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Typhonium flagelliforme inhibits the proliferation of murine leukemia WEHI-3 cells in vitro and induces apoptosis in vivo
Abstract
Typhonium flagelliforme (TF) is a tropical plant, traditionally used by the ethnic population of Malaysia for the cure of various cancers. This plant had shown to induce antiproliferative effect as well as apoptosis in cancer cells. However, there is no available information to address that TF affects murine leukemia cells in vitro and in vivo. Here, we investigated in vitro and in vivo effects of TF on murine leukemia WEHI-3 cells. It was found that the growth of leukemia cells in vitro was inhibited by the various extracts of TF. Among these fractions, the dichloromethane (DCM) tuber extracts of TF showed the lowest IC(50) (24.0 ± 5.2 μg/ml) and had demonstrated apoptogenic effect when observed under fluorescent microscope. We investigated the in vivo effects of DCM tuber extracts of TF on murine leukemia cells, and the results showed that the counts of immature granulocytes and monocytes were significantly decreased in peripheral blood of BALB/c leukemia mice after the oral administration of DCM tuber extracts of TF for 28 days with three doses (200, 400 and 800 mg/kg). These results were confirmed by observing the spleen histopathology and morphology of enlarged spleen and liver in leukemia mice when compared with the control. Furthermore, the cell death mechanism in the spleen tissue of treated mice was found via apoptosis.
... The in vitro studies evaluated cytotoxic activity, apoptotic activity, caspase activity, and telomerase expression levels on WEHI-3, P388, CEM-ss, and Raji cell lines. It was found that T. flagelliforme extract had IC50 ranging between 6 to above 100 μg/mL, with chloroform extract showing the lowest IC50 while DCM extract showed the second-lowest IC50 (Choo, Chan, Sam, et al., 2001;Mohan, Abdul, Abdelwahab, Al-Zubairi, Aspollah Sukari, et al., 2010;Mohan et al., 2008Mohan et al., , 2011Purwaningsih et al., 2017). In addition, T. flagelliforme extract was able to induce apoptosis in leukemia cell lines. ...
... In addition, T. flagelliforme extract was able to induce apoptosis in leukemia cell lines. It was observed through DNA fragmentation, membrane blebbing, nuclear chromatin condensation, and apoptotic body formation, as well as cell shrinkage (Mohan, Abdul, Abdelwahab, Al-Zubairi, Aspollah Sukari, et al., 2010;Mohan et al., 2008Mohan et al., , 2011. ...
... T. flagelliforme extract also significantly reduced the spleen tumor and liver size in a dose-dependent manner. TUNEL assay also confirmed that the spleen treated with T. flagelliforme extract increased the number of apoptotic cells (Mohan, Abdul, Abdelwahab, Al-Zubairi, Aspollah Sukari, et al., 2010). ...
Cancer is a global health issues that can affect anyone. Cancer is treated conventionally with surgery, chemotherapy, radiotherapy, and hormone therapy. However, the high cost of conventional treatments is a burden for cancer patients. Therefore, many cancer patients seek for cheaper yet effective alternative treatments. Typhonium flagelliforme is a taro-like plant that can be found across Indonesia. Numerous researches on the anticancer effect of T. flagelliformehave been conducted. However, a systematic review on the anticancer property of T. flagelliforme is still lacking. Therefore, this review aimed to systematically evaluate the scientific evidence for the anticancer activities in T. flagelliforme. Five databases were used as the search engine using the designated search terms and studies were selected based on the inclusion criteria. The anticancer evaluation in 30 studies selected were conducted in leukemia, lymphoma, breast, oral, cervical, lung, liver, colon, and squamous cell carcinoma. The result showed that T. flagelliforme could inhibit cancer cell proliferation with most of the IC50 are less than 200 μg/mL. T. flagelliformeinduced an increase of caspase-3 and -9, as well as a decrease of the anti-apoptotic Bcl-2 protein expression. The expression of p21 protein was increased after treatment of T. flagelliforme extract while the tyrosine kinase, Ki67, HER2/neu, telomerase, and COX-2 expressions were decreased implying T. flagelliforme could inhibit tumor growth and development. Lastly, T. flagelliforme is also capable in reducing the possibility of cancer cell invasion. Findings suggest that T. flagelliforme is potential to further developed for cancer treatment.
... The tissue sections were covered with 100 ml of equilibration buffer followed by 50 ml of rTdT incubation buffer. The samples were then stained by propidium iodide solution and were analyzed under a fluorescence microscope [24]. ...
... difference in apoptotic positive cells, clearly indicating that thymoquinone has a time-dependent apoptogenic effect. On the other hand, there were no statistically significant (p.0.05) differences in necrotic counts at different times during treatment (24,48, and 72 h) [27]. ...
... In comparison to leukemic control group, thymoquinone (100 mg/kg) induced apoptosis to the spleen and liver tissues. However, normal spleen showed few amount of apoptotic cells [24]. ...
Thymoquinone is an active ingredient isolated from Nigella sativa (Black Seed). This study aimed to evaluate the in vitro and in vivo anti-leukemic effects of thymoquinone on WEHI-3 cells.
The cytotoxic effect of thymoquinone was assessed using an MTT assay, while the inhibitory effect of thymoquinone on murine WEHI-3 cell growth was due to the induction of apoptosis, as evidenced by chromatin condensation dye, Hoechst 33342 and acridine orange/propidium iodide fluorescent staining. In addition, Annexin V staining for early apoptosis was performed using flowcytometric analysis. Apoptosis was found to be associated with the cell cycle arrest at the S phase. Expression of Bax, Bcl2 and HSP 70 proteins were observed by western blotting. The effects of thymoquinone on BALB/c mice injected with WEHI-3 cells were indicated by the decrease in the body, spleen and liver weights of the animal, as compared to the control.
Thymoquinone promoted natural killer cell activities. This compound showed high toxicity against WEHI-3 cell line which was confirmed by an increase of the early apoptosis, followed by up-regulation of the anti-apoptotic protein, Bcl2, and down-regulation of the apoptotic protein, Bax. On the other hand, high reduction of the spleen and liver weight, and significant histopathology study of spleen and liver confirmed that thymoquinone inhibited WEHI-3 growth in the BALB/c mice. Results from this study highlight the potential of thymoquinone to be developed as an anti-leukemic agent.
... The WEHI-3B cell line was originally derived from the BALB/c mice (Warner et al., 1969). It is a good animal model that has been extensively used to induce leukemia in BALB/c mice for evaluation of anti-leukemia effects of drugs and herbal plants (He and Na, 2001; Lin et al., 2009; Mohan et al., 2010; Tsou et al., 2009; Wen et al., 2010; Yu et al., 2010). Supercritical fluid extraction (SFE) is a process to extract bioactive components by using supercritical fluid as the solvent. ...
... Body weight of the mice was recorded daily for 14 days following single oral dose of kenaf seed oil emulsion. The percentage of weight loss (relative to the initial starting weight) was calculated and a loss of greater than 15% is considered toxic (Mohan et al., 2010; Phillips et al., 2004). ...
... However, both populations of the cells decreased after treatment with kenaf seed oil. The reduction might be due to the induction of apoptosis or differentiation of immature cells into the mature one (He and Na, 2001; Mohan et al., 2010). CD11b antigen has a wide tissue distribution such as macrophages, bone marrow, spleen, natural killer cells and blood monocytes and granulocytes. ...
Hibiscus cannabinus (kenaf) seed oil is a rich source of bioactive phytochemicals with high anti-oxidative and cancer chemopreventive properties. However, the seeds are disposed as waste material during the harvesting or processing of kenaf. Preliminary study revealed that kenaf seed oil from supercritical carbon dioxide fluid extraction (SFE) induced apoptosis in WEHI-3B leukemia cells. Thus, this study was carried out to investigate the effects of kenaf seed oil from SFE on WEHI-3B cells in vivo. Acute toxicity study revealed that kenaf seed oil is practically non-toxic by oral route. Treatment with kenaf seed oil increased the population of T cells, but decreased the populations of immature monocytes and granulocytes in the peripheral blood of WEHI-3B/BALB/c mice. The weights of the spleen and liver of WEHI-3B/BALB/c mice decreased after the treatment with kenaf seed oil. Moreover, infiltration of leukemic cells into the splenic red pulp reduced after the treatment. In conclusion, kenaf seed oil reduced the severity of leukemia in WEHI-3B/BALB/c mice. These results provide information to industrialists and farmers to fully utilize and develop kenaf seed oil as a novel bio-health product.
... Water, ethanol, and methanol extracts of garlic have exhibited lethal cytotoxic effects of 50-75% on cells harvested from 3 patients with acute lymphoblastic leukemia (ALL) and three patients with acute myeloid leukemia (AML) [10]. Typhonium flagelliforme (TF), a herbal plant in Malaysia, exhibited cytotoxic effects in P388 [11] and WEHI-3 [12] murine leukemia cells. Further, in vivo studies showed a significant decline in the cell count of immature monocytes and immature granulocytes, when the TF extract was orally administered for 28 days in a BALB/c leukemic mouse model [12]. ...
... Typhonium flagelliforme (TF), a herbal plant in Malaysia, exhibited cytotoxic effects in P388 [11] and WEHI-3 [12] murine leukemia cells. Further, in vivo studies showed a significant decline in the cell count of immature monocytes and immature granulocytes, when the TF extract was orally administered for 28 days in a BALB/c leukemic mouse model [12]. Studies with grape seed extracts reported induction of apoptosis in Jurkat leukemic cells through a process that involves sustained JNK activation and Cip1/p21 up-regulation, culminating in caspase activation [13]. ...
Numerous edible plants have been reported to interfere with the carcinogenic process, and therefore, the regular consumption of these plant products may reduce the risk of developing cancer. We investigated the effect of papaya fruit and leaves on the cell proliferation of Jurkat T-lymphocytic and Daudi B-lymphocytic leukemia cells. Cells were treated with aqueous or methanolic extracts from leaves, skin, pulp, and seeds from green papaya. The papaya fractions were tested for total phenolic content, total flavonoids content, and anti-oxidation activity using chemical assays. Cell proliferation was measured using a WST-1 assay. Our data indicate that methanol and water extracts of seeds and leaves contained higher concentrations of total phenolic compounds and higher anti-oxidation activity than that of extracts from skin and pulp. Both methanol and water extracts from leaves and skin potently inhibited the proliferation of leukemic Jurkat T-cells and Daudi B-cells. However, the effect was more potent on Jurkat T-cells, and the leaf extracts were more effective than that of skin extracts. None of the pulp or seed extracts showed inhibitory activity on leukemic cell proliferation. Although papaya leaves are not consumed as a food, leaf extracts have been used for the treatment of various conditions, including dengue and malaria fevers, gastric ulcers, low platelet counts, and cancers of the breast, lung, and cervix. Our data suggest that the consumption of papaya leaf extracts may also be beneficial in preventing and/or treating lymphocytic cancer. Isolation of active compounds from papaya leaves will also help in developing new drugs for cancer treatment.
... Many studies investigated that rodent tuber has bioactive compounds as anticancer, antiviral, antibacterial, and antioxidant agents [2][3][4]. Rodent tuber was found to possess useful anticancer activities such as in breast, intestine, prostate gland, liver, leukemia, and cervix [2,5,6]. In vitro mutagenesis was carried out using a combination of gamma-ray irradiation and somaclonal variation to increase the bioactive compounds in rodent tuber plants. ...
... Extracts of the rodent tuber plant have been applied to several cancer cells in vitro. Rodent tuber extract with ethanol fraction has been shown to be effective in inhibiting the growth of T47D breast cancer cells [14], inhibiting the proliferation of T4-lymphoblastoid human cancer cells [4,5] and can inhibit the growth of cell cultures of non-small cell lung carcinoma NCI-H23 [2]. The anticancer activity test can be carried out by antiproliferative extract test on the growth of cancer cells using the clonogenic assay method [15,16]. ...
Objective: The objective of this study was to determine the new bioactive compounds through gas chromatography–mass spectrometry analysis and the cytotoxic activity of two rodent tuber mutant plants against breast cancer cells (MCF-7). Methods: The bioactive compounds in rodent tuber mutant plants were successfully increased by somaclonal variation using gamma rays irradiation technique. Further, the cytotoxicity activity of rodent tuber mutant plants was tested on breast cancer cell line (MCF-7) performed by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide assay method. Results: This results study confirmed that the presence of phytochemical composition in the tuber of rodent tuber mutant plants KB 6–1–3–4 and KB 6–9–5 was found six bioactive compounds from fatty acid groups which have the potential as an anticancer compound, such as octadecanoic acid, hexadecanoic acid, hexadecanoic acid methyl ester, 9-octadecanoic acid, linolelaidic acid methyl ester, and butanoic acid. The results showed that extracts from rodent tuber mutant plants had a cytotoxicity effect on MCF-7 cancer cells with half maximal inhibitory concentration (IC50) values that were lower than the control (mother plant). In vitro tests of KB 6–1–3–4 and KB 6–9–5 against MCF-7 cancer cell lines have IC50 values of 12.482 μg/mL and 7.043 μg/mL, respectively, while it had a lower cytotoxicity effect with the IC50 value of control plant was 19.113 μg/mL. The mutant plants of KB 6-9-5 have 3 times more effective than control. Conclusion: The results of this study clearly indicated that rodent tuber mutant plants have shown promising as an anticancer drug on breast cancer.
... Several previous studies using cultured T4 lymphoblastoid and WEHI-3 leukemia cells have reported that Typhonium flagelliforme may inhibit cancer cell proliferation and induce apoptosis. (7)(8)(9) According to a study conducted in Malaysia, rodent tuber may decrease pain, metastases or dissemination of cancer cells, comprising mammary, nasopharyngeal, cervical, prostatic, pancreatic, and pulmonary cancers. (7) In addition, rodent tuber grows abundantly in Indonesia and Malaysia, and has been used for the treatment of hemorrhoids and skin diseases, for neutralizing narcotic drugs, and has antibacterial and antioxidant properties. ...
... (7) In addition, rodent tuber grows abundantly in Indonesia and Malaysia, and has been used for the treatment of hemorrhoids and skin diseases, for neutralizing narcotic drugs, and has antibacterial and antioxidant properties. (8,9) Regarding the chemical compounds of rodent tuber, there is a scarcity of information. Phytochemical analyses of rodent tuber performed at the Research Station for Medicinal and Aromatic Plants (Balai Penelitian Tanaman Obat dan Aromatik) showed the presence of alkaloids, saponins, steroids, and glycosides, but the specific active substances in rodent tuber that play a role in the healing of cancers are as yet unknown. ...
p> Background
Cancer cells have a relatively high telomerase activity compared to normal cells, so that cancer cells have the ability for continued proliferation and uncontrolled mitosis. Telomerase is an enzyme responsible for the length of telomeres, DNA segments located at the ends of eukaryotic chromosomes. Natural materials such as rodent tuber ( Typhonium flagelliforme ) have anticancer potential. The purpose of the present study was to determine the effects of Typhonium flagelliforme extract on telomerase expression in HeLa cervical cancer and T47D breast cancer cells.
Methods
This experimental laboratory study was conducted on cultured HeLa and T47D cancer cell lines, with normal Vero cells as controls, and using RPMI and M199 culture media. The study comprised three groups, i.e. controls, and groups receiving Typhonium flagelliforme extract at doses of ½ IC50 and IC50. Telomerase expression was measured by immunohistochemistry (IHC). Analysis of variance and LSD multiple comparison test were used to analyze the data.
Results
Telomerase expression in cancer cells showed significantly higher values compared to normal Vero cells. Typhonium flagelliforme extract was capable of significantly decreasing telomerase expression in cancer cells receiving the extract.
Conclusion
Typhonium flagelliforme extract at different doses is capable of decreasing telomerase expression more effectively in cervical cancer cells than in breast cancer cells. This study shows that Typhonium flagelliforme may have anti-cancer activity, necessitating further investigations.
... However, only a portion of these plants has been developed for therapeutic drugs [14], particularly cancer therapy [15]. Several plants have been investigated as therapeutic agents for leukemia, lung cancer, breast cancer, liver cancer, and prostate cancer [16][17][18][19][20][21][22][23]. Several anticancer compounds have also been isolated from other sources such as microbes (endophytic bacteria/yeast and fungi). ...
Breast and cervical cancers are the leading cause of death in women, and chemotherapy with cytotoxins is the usual treatment. This study evaluated the cytotoxicity of guava leaf (Psidium guajava L.) extracts as an alternative chemotherapeutic drug. Although many studies related to the cytotoxic effects of guava leaf (Psidium guajava L.) on cancer cells have been reported, the effects of guava leaf fractions on human breast and cervical cancer cells (T47D, MCF-7, and HeLa) have never been evaluated. Herein, we researched candidate activities of ethanol, ethyl acetate, and n-extracts from guava leaf fractions and their effect on various human cancer cell lines (T47D, MCF-7, and HeLa cells). The cytotoxicity test was carried out using the microtetrazolium assay for all fractions. We confirmed and showed the in vitro antitumor activity of guava leaf (Psidium guajava L.) fractions in human breast and cervical cancer cells. We found that the effectiveness of anticancer activity increased from ethanol to ethyl acetate to n-hexane fraction. This work underlines the potential of n-hexane fraction as a chemotherapeutic drug. These novel results have important implications for further isolation, identification, and characterization of Psidium guajava L.-based anti-cancer extracts.
... Despite the fortune Journal of Chemistry available research on the molecular details of TGF-β signaling, little is known regarding how TGF-β and Smad influence CML [60]. One of these realized relationships is that the constitutively active tyrosine kinase produced by the specific Bcr-Abl fusion gene on the Philadelphia chromosome can enhance the resistance of malignant cells to TGF-β1-induced growth inhibition and apoptosis [61]. Previous studies reported ursolic acid and oleanolic acid as antagonists of TGF-β1 binding to its receptor in Balb/c 3T3 cells [62]. ...
As part of our research group’s continuous efforts to find alternative treatments for cancer, the aqueous ethanol extract of Sesbania sesban L. Merr. (SS, Egyptian riverhemp) demonstrated an antileukemic activity against K562 cell line. Bioguided fractionation of SS leaves hydroethanolic extract resulted in the isolation of one new compound (33) named as hederatriol 3-O-β-D-glucuronic acid methyl ester as well as 34 known compounds. Seven compounds ((34), (22), (20), (24), (21), (19), and (35)) showed high antiproliferative effects (IC50 = 22.3, 30.8, 31.3, 33.7, 36.6, 37.5, and 41.5 μM, respectively), while four compounds ((32), (5), (29), and (1)) showed milder activities (IC50 = 56.4, 67.6, 83.3, and 112.3 μM, respectively). A mechanistic study was further carried out on a molecular genetics level against several transcription factors signaling pathways that are incorporated in the incidence of cancer. The results showed that compounds (22) and (21) demonstrated a specific inhibition of Wnt pathway (IC50 = 3.8 and 4.6 μM, respectively), while compound (22) showed a specific inhibition of Smad pathway (IC50 = 3.8 μM). Compound (34) strongly altered the signaling of Smad and E2F pathways (IC50 = 5 μM). The bioactive metabolites were further investigated in silico by docking against several targets related to K562 cell line. The results showed that compounds (22) and (34) exhibited a strong binding affinity towards topoisomerase (docking score = −7.81 and −9.30 Kcal/Mole, respectively). Compounds (22) and (34) demonstrated a strong binding affinity towards EGFR-tyrosine kinase (docking score = −7.12 and −7.35 Kcal/Mole, respectively). Moreover, compound (34) showed a strong binding affinity towards Abl kinase (docking score = −7.05 Kcal/Mole).
... Patient-derived cell xenotransplants in immunodeficient murine model systems provide an experimental platform to test the efficacy of novel therapeutic compounds against human AML cells, but these immunodeficient models are limited by their inability to address the interplay of leukemic blasts with different cells of the immune system (13). In this sense, the WEHI-3 murine cell graft model in immunocompetent Balb/c mice, although it induces a pathology of leukemia that could not fully phenocopy human AML, represents a model where leukemic cells over impose the immune system, establish themselves in the bone marrow and develop until they generate a disease pattern that, in the absence of treatment, invariably leads to death; thus, although it is not without limitations, this leukemia study system remains valid (12,(14)(15)(16)(17). ...
Background/aim:
Although acute myeloid leukemia (AML) has traditionally been considered an oncological emergency and initiation of therapy is believed to be crucial to minimizing disease-related morbidity and mortality, it has also been suggested that a certain delay in treatment has no negative consequences in terms of response, early mortality, or survival. We aimed to determine the effect of administration of sodium caseinate (SC), a salt of casein, the main milk protein, with cytarabine or with daunorubicin on survival in mice with well-established leukemia.
Materials and methods:
To assay the time of establishment of leukemia in the bone marrow, Balb/c mice were inoculated with 2.5×10 5 WEHI-3 cells/mouse and after 3, 6 and 9 days were euthanized. Bone marrow mononuclear cells (BM-MNCs) of the femur were obtained and cultured for 120 h with or without rmIL-3 and cell proliferation was evaluated by the crystal violet technique. Then, the effect of administrating SC-cytarabine or SC-daunorubicin on survival rates of mice with well-established leukemia was assayed. Another group of Balb/c mice was inoculated with WEHI-3 cell and after 10 days mice were treated with SC-cytarabine or SC-daunorubicin for 40 days. Survival rates were recorded daily and in surviving mice, the prevalence of bone marrow proliferation after treatment was assayed by the crystal violet technique.
Results:
The assay on the time of establishment of leukemia shows that in 9 days leukemia cells accumulate in the bone marrow in sufficient quantities to sustain an in vitro culture in the absence of growth factors, and we, thus, used this as a criterion of well-established leukemia. When mice with a burden of leukemic cells of more than 9 days were treated with SC-cytarabine or SC-daunorubicin, this resulted in 55% survival for both treatments, and the proliferation assays showed that the bone marrow retained its normal proliferation capacity.
Conclusion:
SC-cytarabine or SC-daunorubicin treatment prolonged the survival rate of Balb/c mice with a burden of well-established leukemia, and there was no negative impact on bone marrow functionality; however, SC-cytarabine or SC-daunorubicin combination options need to be sought to increase survival beyond 40 days.
... Southeast Asian countries, including India and China, utilize this plant for years as alternative cancer therapy. Typhonium Flagelliforme is a potential health supplement for treatment lung, breast, rectum, liver, prostate, cervical and pancreatic cancer and leukemia [37]- [43]. Characteristics of this plant can be seen on its leaves. ...
Processing data for specific purposes requires an understanding of the data itself.
A special investigation is needed to understand the data. Liquid ChromatographyMass Spectrometry data are the results of material sample examination, which, in
this case, was the sample of Rodent Tuber plants. The data need to be examined
and understood whether they are timeseries or not, which is important for further
processing. In this paper, we examined the data visually with graphs extracted
from the data by human eyes and examination using the Augmented Dickey Fuller
Test conducted by python programming with its library. From human eyes visual
observations and computation using ADF Test, it can be concluded that Liquid
Chromatography-Mass Spectrometry data are stationary timeseries.
... Recent research, indicates a pheophorbide compound isolated from keladi tikus was reported to have an antiproliferation activity on several types of cancerous cells in vitro (Lai et al., 2010). In addition, dichloromethane extract of the tuber parts of keladi tikus proved to have anti-leukemia activity both in vitro and in vivo (Mohan et al., 2010). Despite the pharmacological effects reported, the safety-related information of keladi tikus is still very limited. ...
Keladi tikus (Typhonium flagelliforme (Lodd.) Blume) is an Indonesian medicinal plant that has various pharmacological properties. Zebrafish (Danio rerio) has been proposed as a model that can bridge the gap between cell assays and rodent assays. Evaluation of the toxic effects of natural products using the Zebrafish model can be assessed starting from the blastula stage of embryonic development. This study aims to investigate the potential acute toxicity effect of keladi tikus-ethanol extract (KTEE) using zebrafish embryos. A static non-replacement regime for acute toxicity testing was used. Wild-type zebrafish embryos were exposed to various concentrations of KTEE (50, 100, 200, 400, 800, 1600 µg/mL) starting at 6 hours post-fertilization (hpf) until 96 hpf. The results showed that the survival rate of zebrafish embryos decreased as the concentration of the test extract increased. The LC50 values of KTEE were 494.553 µg/mL at 96 hpf and 555.787 µg/mL at 72 hpf. Embryotoxicity effect of KTEE includes hatching delays and decreased heartrate on zebrafish embryos, especially at high concentrations. KTEE also caused abnormalities in embryo morphology, including pericardial edema, jaw and tail deformity.
... [6] Untreated control of HDF/MCF-7, [7] Pheophorbide 8 µM HDF/ MCF-7 (P8). [4] Andrographolide 48 µM HDF/MCF-7 (A48), [1] Combination of Pheophorbide 2 µM and Andrographolide 12 µM (P2A12), [8] Combination of Pheophorbide 4 µM and Andrographolide 24 µM (P4A24). [2] Combination of Pheophorbide 8 µM and 48 µM Andrographolide (P8A48). ...
The objective of this study was to evaluate the effect of combination of pheophorbide (Thyphonium flagelliforme) and andrographolide (Andrographis paniculata) on breast cancer cell culture of MCF-7 in a laboratory study using a posttest only control group design. human dermis fibroblast (HDF) cells were used as a control. Cell culture was obtained from YARSI University biorepository. Control and experimental treatment was given to six groups for each cell culture, namely control without treatment, P8 (pheophorbide 8 μM), A48 (andrographolide 48 μM), P2A12 (combination pheophorbide 2 μM and andrographolide 12 μM), P4A24 (combination pheophorbide 4 μM and andrographolide 24 μM), and P8A48 (pheophorbide combination 8 μM and andrographolide 48 μM). The treatment was given for 24 h, with three repetitions for each group. Results showed a decrease in cell viability in all treatment groups compared to the control group both in MCF-7 cells and HDF cells. Increasing the dose did not cause an increase in toxicity to MCF-7 cells. The use of a combination of pheophorbide and andrographolide showed anti-cancer effectiveness in MCF-7 breast cancer cell lines at 4 μM pheophorbide doses and 24 μM andrographolide
... Also recent studies have shown relationship between consumption of fruit and vegetable with cancer. The studies have shown some of herbal medicine such as Hibiscus cannabinus (Kenaf) (6), Ginseng root (7), Euphorbia formosana (8), Allium sativum (9), and Vernonia amygdalina (10) have cytotoxic effects on leukaemia cell lines. Ferula species is known for its oleo-resins that are recognized valuable industrial crops. ...
Ferula species known for its oleo-resins that are recognized valuable industrial crops and food products. In this study, we examined the level of cellular oxidants, cytotoxicity, apoptosis and differentiation induced by oleo resin gum from Ferula gummosa (30-250 µg/mL), as well as Arsenic trioxide (50 µM, as positive control), in leukemic (NB4 and HL-60 cells) and normal polymorph nuclear cells during 72 h. Resazurin assay was used to determine cell viability following treatment with F. gummosa (30-250 µg/mL). Intracellular reactive oxygen species was measured by fluorimetry using carboxy 2', 7'-dichlorofluorescein diacetate. Apoptotic cells were evaluated using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). Differentiation of cells evaluated by Giemsa staining and Nitro Blue Tetrazolium reduction. F. gummosa showed a concentration-dependent suppression in cell survival with IC50 values of 41.8 µg/mL for HL60 and 59.2 µg/mL for NB4 cells after 72 h treatment. ROS formation and apoptotic cells were concentration-dependently increased following treatment with F. gummosa, similar to As2O3. F. gummosa did not induce differentiation of leukemic cells towards granulocytic pattern. The resin did not have toxic effect on PMN cells (<800 µg/mL). In conclusion, the present study demonstrated that F. gummosa induced apoptosis through ROS mechanism on leukemic cells as a concentration and time dependent manner. The precise signaling pathway by which F. gummosa induce apoptosis needs further research.
... Moreover, administration of CZ also able to reduces the incidence of mammary gland tumors in rats 10 . In addition, treatment of TF orally at the dose of 200, 400, and 800 mg/kg BW in BALB/c mice leukemia during 28 days was evidenced reduces the number of immature granulocyte cells in the peripheral blood circulation 11 Furthermore, my previous study demonstrated that administration of TF syrup at the dose of 40 and 80 mg/ml once daily for 25 days in C3H mice were inoculated with breast cancer cells, albeit capable of lowering expression of Her2neu protein, but did not reduce the tumor volume. The active substance that mediates PN, CZ, and TF action on tumor including expression of p53, caspase 3 and 7, and reducing immature granulocyte in peripheral blood circulation is polyphenols, curcumin, and flavonoids respectively. ...
Introduction: Breast cancer is the most common cancer and the second leading cause of cancer death in women. Combinations of Thyponium flageliforme, Curcuma zedoaria and Phyllanthus niruri (TCP) have been proven as immuno-modulator and retarded tumor progression in vivo. Objective: to investigate the apoptotic and anti-proliferative effect of TCP in C3H mice suffered from adenocarcinoma cells. Methods: 18 mice inoculated with one million cells of breast adenocarcinoma per mouse were assigned into three groups, TCP with dosage 85 and 125 mg were treated for 21 days to evaluate tumor volume, expression of Ki-67 and Capase3. Expression of Ki-67 and Capase3 were measured by immunohistochemistry staining, whilst cancer volume was assessed using calipers. Results: Mann Whitney analysis indicated that tumor volume in TCP groups were lower, otherwise for expression of caspase3, p < 0.05 respectively. Whereas expression of Ki-67 in TCP groups was lower, but not significant, p > 0.05. Conclusion: TCP treatment with 85 and 125 mg dosage capable of reducing tumor volume, increasing caspase3 and lowering Ki-67 expression.
... They were divided into six groups (0–6), with each containing 10 mice. Group 0 was comprised of healthy controls (not injected with WEHI-3, leukaemic cells); whereas the other five groups (1–5) were injected intraperitoneally (i.p) with WEHI-3 cells (1 × 10 6 cells/mice)[12]using a 1 ml syringe with a 27G needle. Then, 4 days after the inoculation, the mouse blood smear was performed to confirm the induction of leukaemia; following that, the leukaemic mouse group 1 was untreated (Leu), group 2 was treated daily with Tween 20 (the vehicle) (Leu + Tween 20), and groups 3 and 4 were treated with BM 30 mg/ kg(Leu + BM-LD) and 60 mg/kg (Leu + BM-HD), respectively. ...
Background
Beta-mangostin (BM) is a xanthone-type of natural compound isolated from Cratoxylum arborescens. This study aimed to examine the apoptosis mechanisms induced by BM in a murine monomyelocytic cell line (WEHI-3) in vitro and in vivo.
Methods
A WEHI-3 cell line was used to evaluate the cytotoxicity of BM by MTT. AO/PI and Hoechst 33342 dyes, Annexin V, multiparametric cytotoxicity 3 by high content screening (HCS); cell cycle tests were used to estimate the features of apoptosis and BM effects. Caspase 3 and 9 activities, ROS, western blot for Bcl2, and Bax were detected to study the mechanism of apoptosis. BALB/c mice injected with WEHI-3 cells were used to assess the apoptotic effect of BM in vivo.
Results
BM suppressed the growth of WEHI-3 cells at an IC50value of 14 ± 3 μg/mL in 24 h. The ROS production was increased inside the cells in the treated doses. Both caspases (9 and 3) were activated in treating WEHI-3 cells at 24, 48 and 72 h. Different signs of apoptosis were detected, such as cell membrane blebbing, DNA segmentation and changes in the asymmetry of the cell membrane. Another action by which BM could inhibit WEHI-3 cells is to restrain the cell cycle at the G1/G0 phase. In the in vivo study, BM reduced the destructive effects of leukaemia on the spleen and liver by inducing apoptosis in leukaemic cells.
Conclusion
BM exerts anti-leukaemic properties in vitro and in vivo.
... Scientific study had documented the medicinal value of T. flagelliforme for anticancer [4], antibacterial [5], anti-inflammatory and antioxidant properties [5]. Since this species is well recognized as important medicinal plants, it is sought after for further research and uses. ...
Objective: To determine the effects of different strength of Murashige and Skoog (MS) media (full, ½ and ¼) in solid and liquid media on in vitro growth of Typhonium flagelliforme (T. flagelliforme), whereby an optimum media composition can be provided for mass propagation of T. flagelliforme.
Methods: Rhizome bud of T. flagelliforme was obtained from the axenic in vitro established T. flagelliforme plantlets in Plant Tissue Culture Laboratory, Universiti Teknologi MARA, Shah Alam. Rhizome bud was used as explant and cultured onto shoot proliferation medium under different strength of MS media (full, ½, ¼) in solid and liquid culture media.
Results: After 6 weeks of culture, the number of shoot, number of leaf, number of root, height of shoot, fresh weight, dry weight and chlorophyll content of T. flagelliforme were analyzed. A comparison was made between liquid and solid culture media. The results revealed that the liquid culture media were more effective for all the growth parameters (shoot height, shoot number, leaf number, root number, fresh weight, dry weight, chlorophyll a and chlorophyll b content) compared to solid culture media. Apart from that, this study revealed the positive relationship between strength of MS media and type of culture media (solid and liquid media) to the growth of T. flagelliforme. Growth of T. flagelliforme was improved when MS strength was increased in liquid media. In contrast, growth of T. flagelliforme was improved when MS strength was decreased in solid media.
Conclusions: Through this study, an optimum media composition for mass propagation of T. flagelliforme had been established by observing effects of MS media strength and type of culture media (solid and liquid media) on the growth of T. flagelliforme.
... In Malaysia, studies on plants and herbs as potential sources of cancer therapeutics are on the rise. Among the plants being investigated for their therapeutic potential include Typhonium flagelliforme for treatment of leukemias [10,11], chalcone from Boesenbergia rotunda for lung cancers [12], and Elephantopus scaber for human breast cancers [13]. At the same time, several active principles have been identified and investigated to include girinimbine from roots of Murraya koenigii for liver cancers [14], dentatin from wild shrub Clausena excavata Burm for prostate cancers [15], kenaf seed oil from Hibiscus cannabinus and phenylbutenoids from Zingiber cassumunar Roxb for leukemias [16,17]. ...
Cancer is the major killer and dreaded disease caused by abnormal proliferation of body cells. These abnormal cells can block circulations, and damage organ functions which may lead to death. The annual incidence of cancer worldwide is estimated at 30,000. While the most common killer among malignancies is lung cancer, breast cancer poses the biggest threat. With the increasing prevalence of cancer, patients are now turning to complementary and alternative medicine (CAM) to treat the disease.
... Bioactive compounds of rodent tuber are alkaloid, saponin [2], terpenoid, steroid [3], flavonoid, and glycoside [4]. Rodent tuber has anticancer activities against cancer of liver [5], breast, colon, prostate gland, cervix [2], and leukemia [6]. Rodent tuber has been propagated conventionally by tuber separation method. ...
Rodent tuber is an Indonesian anticancer herbal plant whose genetic variation is low. Genetic variation is crucial to be increased to increase the content of anticancer bioactive compounds. In vitro calli of rodent tuber had been mutated by gamma irradiation. Mutant plants had their genome modified and therefore also affected protein expression. The purpose of this research is to identify protein expression differences between mutant and control rodent tuber plants by 1D and 2D PAGE. 1D PAGE had shown differences in the number of protein bands between control and mutant. M6/2 did not show any expression of 6,9 kD protein. 2D PAGE had identified 24 proteins which were expressed by control and M6/2 plants. Among them, 13 spots were up-regulated and 9 spots were down-regulated in M6/2 plants. In conclusion, 1D and 2D PAGE analyses had successfully proven the changes in protein expression between control and mutant rodent tuber plants.
... Bioactive compounds in this plant arealkaloids, saponins, steroids, and glycosides [3]. Rodent tuber's extract was cytotoxic against the cancer of lung, breast [4], liver [5], blood(leukemia) [6], colon, prostate gland, and cervix [7]. Its hexane extract was cytotoxic against Artemia salina larvae [8]. ...
Rodent tuber (Typhonium flagelliforme Lodd.) is an Indonesian anticancer medicinal plant. The natural genetic diversity of rodent tuber is low due to vegetative propagation. Plant's genetic diversity has to be increased for obtaining clones which contain a high amount of anticancer compounds. In vitro calli were irradiated with 6 Gy of gamma ray to produce in vitro mutant plantlets. Mutant plantlets were acclimated and propagated in a greenhouse. This research was aimed to identify the chemical compounds in the leaves and tubers ofthe fourth generation of rodent tuber's vegetative mutant clones (MV4) and control plantsby using GC- MS method. Leaves and tubers of MV4 each contained 2 and 5 anticancer compounds which quantities were higher compared to control plants. MV4 leaves contained 5 new anticancer compounds while its tubers contained 3 new anticancer compounds which were not found in control. The new anticancer compounds in leaves were hexadecanoic acid, stigmast-5-en-3-ol, ergost-5-en-3-ol, farnesol isomer a, and oleic acid while the new anticancer compounds in tubers were alpha tocopherol, ergost-5-en-3-ol, and beta-elemene. Rodent tuber mutant clones are very potential to be developed into anticancer drugs.
... In Malaysia, studies on plants and herbs as potential sources of cancer therapeutics are on the rise. Among the plants being investigated for their therapeutic potential include Typhonium flagelliforme for treatment of leukemias [10,11], chalcone from Boesenbergia rotunda for lung cancers [12], and Elephantopus scaber for human breast cancers [13]. At the same time, several active principles have been identified and investigated to include girinimbine from roots of Murraya koenigii for liver cancers [14], dentatin from wild shrub Clausena excavata Burm for prostate cancers [15], kenaf seed oil from Hibiscus cannabinus and phenylbutenoids from Zingiber cassumunar Roxb for leukemias [16,17]. ...
Cancer is the major killer and dreaded disease caused by
abnormal proliferation of body cells. These abnormal cells can
block circulations, and damage organ functions which may
lead to death. The annual incidence of cancer worldwide is
estimated at 30,000. While the most common killer among
malignancies is lung cancer, breast cancer poses the biggest
threat.
... Anticancer compounds can be found in all parts of a rodent tuber plant, including root, tuber, stem, and leaf [3]. Rodent tuber has cytotoxicity against cancer of lung, breast [4], liver [5], leukemia [6], colon, prostate, and cervix [7]. Rodent tuber's extract has also been proven to be able to prevent breast and cervical cancer [8]. ...
Rodent tuber (Typhonium flagelliforme Lodd.) is an herbal medicinal plant with anticancer activity. The genetic diversity of rodent tuber is low due to vegetative propagation. Somatic cell population of rodent tuber from Bogor had been irradiated with gamma ray to increase genetic diversity. There were 37 clones of first generation putative mutant (MV1) which had been analyzed based on morphological and RAPD markers. Out of those 37 MV1 clones, there were 17 clones which had undergone genetic mutation and had a diversified genetic profile. MV1 had been regenerated to fourth generation putative mutant clones (MV4). This research was aimed to analyze the mutation stability of MV4 based on morphological and RAPD markers. Clone 6-1-2 had the highest increase of the number of shoots and leaves than control and the other MV4 clones, with 4.7 and 19.7 shoots and leaves, respectively. Clone 6-1-1-6 obtained the highest increase in plant height than control and the other MV4 clones, i.e. 25.2 cm. Clone 6-9-5 had the weightiest fresh and dry weight, i.e. 41.67 gram and 12.01 gram respectively. RAPD molecular marker analysis of MV4 by using 15 primers had produced 64 polymorphic DNA bands out of 146 total bands. OPD-10 primer produced the highest number of polymorphic bands, i.e. 15 polymorphic bands out of 17 total bands with sizes 200-2000 bp. RAPD profile of MV4 had showed 5 main clusters at similiarity coefficient cut-off 0.91. Morphological characterization and RAPD analysis had proved the stability of genetic mutation of MV4.
... Treatment strategy of leukemia is a multidisciplinary effort. The modalities of treatment include radiotherapy, chemotherapy, immunotherapy and symptomatic and supportive therapy (Syam et al., 2010). Chemotherapy is the major form of treatment for leukemia. ...
Leukemia is the common cause of death and worldwide incidence of this disease is increasing. Chemotherapy is the first choice for leukemia treatment, but the major limitations of standard therapy are its side effects and poor patient compliances. Therefore it is imperative to look for a therapeutic system with lesser side effects urgently to address the underlying causes of poor treatment outcomes. In such a scenario transdermal route for delivery of chemotherapy could be a better alternative that could provide sustained drug level, enhanced activity, self administration and better patient compliances. The present work is focus on the design of nanolipid based transdermal carrier, deformable liposomes bearing cytarabine as a model drug for effective delivery of drug with enhanced transdermal flux. Developed nanocarriers were characterized for their size, morphology, entrapment efficiency, skin penetration and irritation. It could be concluded that nanodeformable liposomes attenuated transdermal flux of cytarabine and could provide a new strategy for leukemia.
... Rodent tuber (Typhonium flagelliforme Lodd.) is a herbal plant from the Araceae family which is widely cultivated in India, Australia, Sri Lanka, Indonesia, and the other a Asian countries with moderate temperature 1 . Rodent tuber has a broad array of bioactive compounds with anticancer activity against lung, breast 2 , liver 3 , blood 4 , intestine, prostate gland, and cervical cancer 5 based on in vitro experiments. Anticancer compounds can be found in all parts of rodent tuber plant, including root, tuber, stem, and leaf 6 . ...
Rodent tuber (Typhonium flagelliforme Lodd.) is an Indonesian native plant which has a high potency to be used as anticancer medicine, but its genetic variation in Indonesia is low. Gamma irradiation can be used to increase genetic diversity. The obj ective of this research is to obtain in vitro culture of the first generation (MV 1) mutants of rodent tuber which had been irradiated by gamma rays and detect genetic changes by RAPD molecular markers. Irradiation was performed using 60 Co to obtain LD 50 value and induce mutation. Calli were then regenerated to obtain MV 1 plantlets and genetic changes were detected by RAPD molecular markers. Plantlets were successfully regenerated from 9 clumps of calli and produced 59 plantlets at 6 Gy. Genetic changes of 11 plantlets from calli were detected by 10 RAPD primers, which produced 69 fragments from 11 mutant plantlets. OPE-20 and OPC-5 primer showed the highest genetic changes with 11 and 8 polymorphic fragments respectively. Nine polymorphic primers showed genetic changes between 250 bp and 2 500 bp.This research had successfully induced mutation, regenerated, and detected genetic changes in 11 mutant plantlets from calli.
... Rodent tuber is a medicinal plant. Several countries in Asia has been using the plant to treat cancer (Chan et al., 2005), such as liver cancer (Lai et al., 2008), breast cancer, intestinal cancer, prostate cancer, cervix cancer (Syahid, 2008), and leukemia (Mohan et al., 2010). Sianipar, Maarisit and Valencia (2013) had reported the toxicity effect rodent tuber's extract and hexane fraction on Artemia salina. ...
Rodent tuber (Typhonium flagelliforme Lodd.) is an Indonesian plant with high medicinal potential as anti-cancer. This plant has a low genetic variation in Indonesia. Gamma irradiation can be used to increase genetic variation. This research aimed to explore the effects of gamma irradiation and somaclonal variation on several mutant rodent tuber lines. Six somatic cell populations, which were treated by 6 Gy of gamma irradiation, were successfully regenerated into plantlets. Six mutant lines had been sub cultured into second and third generations by using optimal regeneration media (MS0 supplemented with 0.5 mg/L BAP and 1 mg/L NAA). Rooting of in vitro plantlets had been done by using optimal rooting media MS0 supplemented with 0 mg/L NAA and 0.5 mg/L NAA). Plantlets with good roots were acclimatized and transferred into greenhouse. The morphology of first generation mutant in greenhouse (M1) were characterized and analyzed by using descriptive statistical method. The observed morphological characters including plant height, shoot number, leaf number, and leaf area. A hundred and seventy five (175) mutant lines were obtained on the third generation (MV3) from five somatic cell populations. Mutant line 6-3-x has the highest mean number of shoots, which was 4.62 shoots. Percentage of plant alive after acclimatization in the greenhouse was 78%. Out of 37 M1 mutant lines, morphologically diverse lines were observed with the highest plant height increase: 15.5 cm on 6-2-4-1 mutant line, the highest plant shoots increase: 5 shoots on 6-6-7-8 mutant line, the highest leaf number increase: 17 leaves on 6-6-7-8 mutant line, and the highest leaf area increase: 47.24 cm 2 on 6-2-5-2 mutant line. In conclusion, gamma irradiation and somaclonal variation could increase genetic variation of mutant rodent tuber lines as shown by morphological data.
... Phase-contrast micrographs reveal that SOE induced typical morphological characteristics of apoptosis, including cell shrinkage, apoptotic vacuoles, membrane blebbing and formation of floating cells, in a dose-and time-dependent manner. These changes are in agreement with findings in literature on apoptosis [21][22][23]. ...
Endometrial cancer is a common malignancy of the female genital tract. This study demonstrates that Siegesbeckia orientalis ethanol extract (SOE) significantly inhibited the proliferation of RL95-2 human endometrial cancer cells. Treating RL95-2 cells with SOE caused cell arrest in the G2/M phase and induced apoptosis of RL95-2 cells by up-regulating Bad, Bak and Bax protein expression and down-regulation of Bcl-2 and Bcl-xL protein expression. Treatment with SOE increased protein expression of caspase-3, -8 and -9 dose-dependently, indicating that apoptosis was through the intrinsic and extrinsic apoptotic pathways. Moreover, SOE was also effective against A549 (lung cancer), Hep G2 (hepatoma), FaDu (pharynx squamous cancer), MDA-MB-231 (breast cancer), and especially on LNCaP (prostate cancer) cell lines. In total, 10 constituents of SOE were identified by Gas chromatography-mass analysis. Caryophyllene oxide and caryophyllene are largely responsible for most cytotoxic activity of SOE against RL95-2 cells. Overall, this study suggests that SOE is a promising anticancer agent for treating endometrial cancer.
... Choo et al. [60] performed an experiment which proved cytotoxic activity on murine P388 leukaemia cell line. Cytotoxicity studies of T. flagelliforme were also performed in vitro against human T3-lymphoblastoid cell lines (CEM-ss) and significant effects were observed [37]. They also further investigated the effects of the leaves extract in in vivo study using BALB/c leukaemic mice model and found that there were significant reductions in the cell count of immature granulocytes and monocytes when the TF extract is orally administered for 28 days at different doses of 200, 400, and 800 mg/kg. ...
In developing countries, herbal therapy is the first and basis form of treatment for most types of diseases. About 75–80% of the world’s population prefers herbal therapy as a major treatment due to its better adequacy and satisfactoriness, which enhance human body’s symmetry with minimal side effects. Fruits and plants have been presented from the past as promising tools in becoming a natural anticancer agents. Many of these plant extracts are currently used in cancer therapy and prevention. This review paper will particularly explore and emphasize on herbs and fruits used in the treatment of the leukaemia.
... These findings are similar to those reported in previous BALB/c mice leukemia model studies. 12,35 Transmission electron microscopy TEM is a useful analytical tool in cell morphological studies in cancers. In this study, the spleen from untreated control mice showed normal cellular features. ...
Cancer nanotherapy is progressing rapidly with the introduction of many innovative drug delivery systems to replace conventional therapy. Although the antitumor activity of zerumbone (ZER) has been reported, there has been no information available on the effect of ZER-loaded nanostructured lipid carrier (NLC) (ZER-NLC) on murine leukemia cells. In this study, the in vitro and in vivo effects of ZER-NLC on murine leukemia induced with WEHI-3B cells were investigated. The results from 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, Hoechst 33342, Annexin V, cell cycle, and caspase activity assays showed that the growth of leukemia cells in vitro was inhibited by ZER-NLC. In addition, outcomes of histopathology, transmission electron microscopy, and Tdt-mediated dUTP nick-end labeling analyses revealed that the number of leukemia cells in the spleen of BALB/c leukemia mice significantly decreased after 4 weeks of oral treatment with various doses of ZER-NLC. Western blotting and reverse-transcription quantitative polymerase chain reaction assays confirmed the antileukemia effects of ZER-NLC. In conclusion, ZER-NLC was shown to induce a mitochondrial-dependent apoptotic pathway in murine leukemia. Loading of ZER in NLC did not compromise the anticancer effect of the compound, suggesting ZER-NLC as a promising and effective delivery system for treatment of cancers.
... These findings are similar to those reported in previous BALB/c mice leukemia model studies. 12,35 Transmission electron microscopy TEM is a useful analytical tool in cell morphological studies in cancers. In this study, the spleen from untreated control mice showed normal cellular features. ...
Cancer nanotherapy is progressing rapidly with the introduction of many innovative drug delivery systems to replace conventional therapy. Although the antitumor activity of zerumbone (ZER) has been reported, there has been no information available on the effect of ZER-loaded nanostructured lipid carrier (NLC) (ZER-NLC) on murine leukemia cells. In this study, the in vitro and in vivo effects of ZER-NLC on murine leukemia induced with WEHI-3B cells were investigated. The results from 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, Hoechst 33342, Annexin V, cell cycle, and caspase activity assays showed that the growth of leukemia cells in vitro was inhibited by ZER-NLC. In addition, outcomes of histopathology, transmission electron microscopy, and Tdt-mediated dUTP nick-end labeling analyses revealed that the number of leukemia cells in the spleen of BALB/c leukemia mice significantly decreased after 4 weeks of oral treatment with various doses of ZER-NLC. Western blotting and reverse-transcription quantitative polymerase chain reaction assays confirmed the antileukemia effects of ZER-NLC. In conclusion, ZER-NLC was shown to induce a mitochondrial-dependent apoptotic pathway in murine leukemia. Loading of ZER in NLC did not compromise the anticancer effect of the compound, suggesting ZER-NLC as a promising and effective delivery system for treatment of cancers.
... These findings are similar to those reported in previous BALB/c mice leukemia model studies. 12,36 Transmission electron microscopy TEM is a useful analytical tool in cell morphological studies in cancers. In this study, the spleen from untreated control mice showed normal cellular features. ...
Cancer nanotherapy is progressing rapidly with the introduction of many innovative drug delivery systems to replace conventional therapy. Although the antitumor activity of zerumbone (ZER) has been reported, there has been no information available on the effect of ZER-loaded nanostructured lipid carrier (NLC) (ZER-NLC) on murine leukemia cells. In this study, the in vitro and in vivo effects of ZER-NLC on murine leukemia induced with WEHI-3B cells were investigated. The results from 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, Hoechst 33342, Annexin V, cell cycle, and caspase activity assays showed that the growth of leukemia cells in vitro was inhibited by ZER-NLC. In addition, outcomes of histopa-thology, transmission electron microscopy, and Tdt-mediated dUTP nick-end labeling analyses revealed that the number of leukemia cells in the spleen of BALB/c leukemia mice significantly decreased after 4 weeks of oral treatment with various doses of ZER-NLC. Western blotting and reverse-transcription quantitative polymerase chain reaction assays confirmed the antileukemia effects of ZER-NLC. In conclusion, ZER-NLC was shown to induce a mitochondrial-dependent apoptotic pathway in murine leukemia. Loading of ZER in NLC did not compromise the anti-cancer effect of the compound, suggesting ZER-NLC as a promising and effective delivery system for treatment of cancers.
... Choo et al. [60] performed an experiment which proved cytotoxic activity on murine P388 leukaemia cell line. Cytotoxicity studies of T. flagelliforme were also performed in vitro against human T3-lymphoblastoid cell lines (CEM-ss) and significant effects were observed [37]. They also further investigated the effects of the leaves extract in in vivo study using BALB/c leukaemic mice model and found that there were significant reductions in the cell count of immature granulocytes and monocytes when the TF extract is orally administered for 28 days at different doses of 200, 400, and 800 mg/kg. ...
In developing countries, herbal therapy is the first and basis form of treatment for most types of diseases. About 75–80% of the world’s population prefers herbal therapy as a major treatment due to its better adequacy and satisfactoriness, which enhance human body’s symmetry with minimal side effects. Fruits and plants have been presented from the past as promising tools in becoming a natural anticancer agents. Many of these plant extracts are currently used in cancer therapy and prevention. This review paper will particularly explore and emphasize on herbs and fruits used in the treatment of the leukaemia.
... Liver diseases such as hepatocellular carcinoma, viral and alcoholic hepatitis and non-alcoholic steatosis are the most common liver-associated and prevalent diseases in the World, which are closely associated with jaundice [12]. As modern medicine fails to cure these ailments completely and harmlessly, research on natural products has gained much attention in the drug discovery against diseases such as liver diseases [13]. Most of the time the research will be guided by some traditional practice or from undocumented treatment modalities [14,15]. ...
In the Indian system of traditional medicine (Ayurveda) it is recommended to consume Ipomoea aquatica to mitigate disorders like jaundice. In this study, the protective effects of ethanol extract of I. aquatica against liver damage were evaluated in thioacetamide (TAA)-induced chronic hepatotoxicity in rats. There was no sign of toxicity in the acute toxicity study, in which Sprague-Dawley (SD) rats were orally fed with I. aquatica (250 and 500 mg/kg) for two months along with administration of TAA (i.p injection 200 mg/kg three times a week for two months). The results showed that the treatment of I. aquatica significantly lowered the TAA-induced serum levels of hepatic enzyme markers (ALP, ALT, AST, protein, albumin, bilirubin and prothrombin time). The hepatic content of activities and expressions SOD and CAT that were reduced by TAA were brought back to control levels by the plant extract supplement. Meanwhile, the rise in MDA level in the TAA receiving groups also were significantly reduced by I. aquatica treatment. Histopathology of hepatic tissues by H&E and Masson trichrome stains displayed that I. aquatica has reduced the incidence of liver lesions, including hepatic cells cloudy swelling, infiltration, hepatic necrosis, and fibrous connective tissue proliferation induced by TAA in rats. Therefore, the results of this study show that the protective effect of I. aquatica in TAA-induced liver damage might be contributed to its modulation on detoxification enzymes and its antioxidant and free radical scavenger effects. Moreover, it confirms a scientific basis for the traditional use of I. aquatica for the treatment of liver disorders.
Leukemia or blood cancer was initially discovered in 1845 and this malignancy was reported in patients who had an amplified number of blood cells, in particular, white blood cells (WBC), due to this disease. The event of leukemia was further identified as a malignant hematopoietic disorder due to both uncontrolled and unlimited proliferation in combination with lack of differentiation of the leukemic stem cells.Furethermore, 75 to 80% of the global population use herbal remedies as primary therapy, mainly because of their better efficiency and satisfaction, which elevate the human body symmetry with the minimum unwanted adverse effects. For the control of cancer, plant products, and fruits have been considered promising tools and are being consumed for centuries. Several plant extracts are also being used for the therapy and prevention of different types of known cancers. Indole-3-carbinol (I3C) is a natural material obtained from Brassica diversity of vegetables and has been reported to serve as a promising cancer preventative agent. In the present review, theauthors mainly tried to focus and emphasize I3C applications in the leukemia treatment.
Superior mutant rodent tuber plant ( Typhoniumflagelliforme ) is a medicinal herb of Indonesia, which immensely useful for anticancer activity. Some studies reported that the leaves and the tubers, conventionally parts of rodent tuber plant, showed various anticancer, antimicrobial, and antioxidant potential. This study aim is to determine the bioactive compounds of superior mutant rodent tuber plants through n -hexane and ethyl acetate fractions using GC-MS for the analysis. Phytochemical characterization of the superior mutant rodent tuber plant extracts was detected by qualitative analysis. A total of 20 bioactive compounds were obtained in an n-hexane fraction. A total of 4 bioactive compounds were identified in ethyl acetate fraction. The GC-MS analysis showed the presence of the major compound in ethyl acetate and n-hexane fraction were 9,11-octadecadienoic acid, methyl ester (55.37%), 9-octadecenoic acid (Z)-, methyl ester (26.77%), hexadecanoic acid, methyl ester (2.97%), 9,12-octadecadienoyl chloride (2.47%), humulene (1.11%), octahydronapthalene (0.97%), alloaromadendrene oxide-(2) (0.85%), pentadecanoic acid, methyl ester (0.80%), methyl tetradecanoate (0.79%), and eucalyptol (0.69%). Most of the identified compounds in the n -hexane and ethyl acetate fractions exhibit following biology activity, such as anticancer, antioxidant, anti-androgenic, antimicrobial, antifungal, and anti-inflammatory. This study provides an overview of the chemical compounds and their beneficial impact on developing drugs from n-hexane and ethyl acetate fraction of the superior mutant rodent tuber plant.
Leukemia is a leukocyte cancer that is characterized by anarchic growth of immature immune cells in the bone marrow, blood and spleen. There are many forms of leukemia, and the best course of therapy and the chance of a patient’s survival depend on the type of leukemic disease. Different forms of drugs have been used to treat leukemia. Due to the adverse effects associated with such therapies and drug resistance, the search for safer and more effective drugs remains one of the most challenging areas of research. Thus, new therapeutic approaches are important to improving outcomes. Almost half of the drugs utilized nowadays in treating cancer are from natural products and their derivatives. Medicinal plants have proven to be an effective natural source of anti-leukemic drugs. The cytotoxicity and the mechanisms underlying the toxicity of these plants to leukemic cells and their isolated compounds were investigated. Effort has been made throughout this comprehensive review to highlight the recent developments and milestones achieved in leukemia therapies using plant-derived compounds and the crude extracts from various medicinal plants. Furthermore, the mechanisms of action of these plants are discussed.
Cancer is still an epidemiological disease in Indonesia. Drug development against cancer still relies to pharmacological laboratories and natural chemicals, which could have side effects. Cancer drug development has entered the stage of molecular biology, where the interaction of ligand chemical structure with receptor protein can be studied with high accuracy. Various chemical compounds, ranging from synthetic, semi-synthetic, to natural materials, developed for the purpose to fight one of the most dangerous diseases. In the context of the development of herbal-based drugs, there has been found heaps of natural compounds, curated and annotated, in various databases belonging to China, Taiwan, Indonesia, Japan, and several other countries. However, problems arise when choosing the best bioactive compounds to develop against cancer. Complexity arises because the metabolic pathway of cancer is very diverse, depending on the type and phase of cancer. Therefore, in this systematic review, we developed a machine learning approach to screen for these bioactive compounds, then took the best candidates for molecular simulation operations that would be tested for validity in wet experiments. Thus, the automation of the candidate drug development process for cancer could be achieved with great significance. It is known that the most effective and efficient machine learning method was Naïve Bayes, but the best in processing large amounts of compound data was classfier SVM. The future of complex bioactive compounds data could be secured by employing deep learning method.
ABSTRAK Penyakit kanker merupakan masalah epidemiologis di tanah air. Pengembangan obat modern sejauh ini masih bergantung pada laboratorium farmakologi dan kimia bahan alam yang dapat memiliki efek samping. Namun, pengembangan obat untuk penyakit kanker sudah memasuki tahap biologi molekuler, dimana interaksi struktur kimia ligan dengan protein reseptor dapat dikaji dengan ketelitian tinggi. Berbagai tipe senyawa kimia, mulai dari sintetik, semi sintetik, hingga bahan alam, dikembangkan untuk keperluan melawan penyakit yang dianggap paling berbahaya tersebut. Dalam konteks pengembangan obat berbasis herbal, telah ditemukan senyawa bahan alam dalam jumlah banyak. Data tersebut dikurasi dan dianotasi oleh basis data milik China, Taiwan, Indonesia, Jepang, maupun beberapa negara lainnya. Hanya saja, permasalahan timbul ketika memilih senyawa bioaktif terbaik untuk dikembangkan untuk melawan penyakit kanker. Kompleksitas timbul karena jalur metabolik penyakit kanker sangat beragam, tergantung pada tipe dan fasenya. Oleh karena itu, dalam telaah sistematis ini, disajikan telaah pendekatan machine learning untuk melakukan penapisan terhadap pustaka senyawa bioaktif tersebut, kemudian proses seleksi kandidat yang terbaik untuk operasi simulasi molekuler, dan selanjutnya teruji validitasnya pada wet experiment. Sehingga proses automatisasi pengembangan kandidat obat bagi penyakit kanker dapat dicapai dengan sangat signifikan. Diketahui bahwa metode machine learning paling efektif dan efisien adalah Naïve Bayes. Namun yang terbaik dalam mengolah data senyawa dalam jumlah besar adalah SVM classifier. Kedepannya, metode deep learning sangat menjanjikan untuk komputasi data senyawa bioaktif yang kompleks.
Kata kunci: machine learning, pengembangan obat, senyawa bahan alam, jalur metabolik, penyakit kanker
Rodent tuber plant (Typhonium flagelliforme Lodd.), as one of the most potent medicinal plants, has to be developed as an active ingredient of degenerative drugs, including cancer drugs. However, this enormous potential must be supported by sustainable cultivation of the plant. The conventional preservation of rodent tuber can be done by planting various accessions in the field, but it will need land availability and intensive plant maintenance. Preservation through in vitro culture is an alternative method that can be used. The aim of this study was to determine the effect of paclobutrazol in suppressing the growth of rodent tuber cultures, and test the ability of culture regeneration after being preserved with paclobutrazol. The media formulation used was Murashige and Skoog (1962) (MS) with the addition of paclobutrazol at concentrations of 4, 5, 6, and 7 mg L-1. The results showed that using paclobutrazol at 5 mg L-1 was the best concentration that can inhibit the elongation of buds, seedling formation, leaf formation, and root elongation until 5 months. Cultures of paclobutrazol treatment at 5 mg L-1 had shoot heights, number of shoots, number of leaves, and root lengths of 0.49, 2.33, 7.23, and 0.3 cm, respectively. Paclobutrazol could inhibit the growth of in vitroculture of rodent tuber and prolong the shelf life of culture up to 5 months. The culture of rodent tuber from paclobutrazol treatment had normal growth and regenerative ability after transfer to the medium regeneration.
The low-dose cytarabine, aclarubicin and granulocyte-colony stimulating factor (G-CSF) (CAG) priming regimen is an effective treatment for patients with relapsed or refractory acute myeloid leukemia (AML) and advanced myelodysplastic syndrome (MDS). G-CSF influences the bone marrow microenvironment (BMM) by mobilizing regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs), as well as by reducing the expression of stromal cell-derived factor-1α (SDF-1α). In the present study, a WEHI-3-grafted BALB/c mouse AML model (AML-M4) was employed to determine how the BMM was altered by different treatment regimens. It was evident that CAG regimen decreased and increased the proportion of Tregs and MDSCs in the bone marrow and spleen, respectively. Furthermore, the CAG regimen downregulated SDF-1α levels in the bone marrow and peripheral blood. However, hematoxylin and eosin staining of the main organs revealed that leukemic cells infiltrated the liver following treatment with the CAG regimen. The present study indicates that the CAG regimen has a positive effect on the immunosuppressive microenvironment in AML and relieves AML-associated BMM immune suppression by decreasing Tregs and MDSCs in the bone marrow and downregulating the SDF-1α/CXCR4 axis in the bone marrow and peripheral blood
Malaysia hosts a diverse range of medicinal plants having therapeutic values including anticancer properties. Over the years, a group of microorganisms called endophytes producing bioactive compounds similar to their host plants has been discovered. These endophytes, producing important compounds such as Taxol, L-asparaginase, and sclerotiorin, can be isolated and cultured in large scale to produce valuable compounds. This alternative approach of producing bioactive compounds is more sustainable than plants, as endophytes are renewable sources. In this chapter, endophytes from Malaysian medicinal plants have been discussed, highlighting the diversity of endophytes, the valuable compounds produced, the current methods used in biosourcing of endophytes, and the future prospects of anticancer agents derived from endophytes of Malaysian medicinal plants.
Objective: To determine the inhibition activity of Typhonium flagelliforme Lodd. Blume (T. flagelliforme) leaf extract on cyclooxygenase 2 (COX-2) expression of colon cancer cells.
Methods: T. flagelliforme leaf extract was prepared to macerate in ethyl acetate. In vitro anticancer activity was assayed by MTT method on WiDr colon cancer cells. This study applied apoptosis induction assay to investigate the mechanism of cell death using double staining method. COX-2 expression was stained by immunocytochemistry.
Results: T. flagelliforme showed anticancer activity and induced apoptosis on WiDr cells through inhibition of COX-2 expression with IC50 70 μg/mL.
Conclusions: This study showed that T. flagelliforme is a promising chemopreventive agent for colon cancer through COX-2 inhibition.
Glioblastoma multiforme (GBM) is a highly malignant brain tumor with limited therapeutic options, and high recurrence and mortality rates. In this study, the anti-tumorigenic properties of luteolin on GBM were examined in vitro. Luteolin was isolated from the whole plant of Glossogyne tenuifolia and the chemical structure was determined by its spectroscopic data. The inhibitory effect of luteolin on the proliferation of GBM 8401 and U87 human glioblastoma cells and the apoptotic effect of this compound on GBM 8401 cells were investigated. Luteolin inhibited the growth of GBM 8401 and U87 cells in a dose- and time-dependent manner. It significantly induced S and G2/M phase cell cycle arrest and apoptotic cell death by decreasing the expression of cyclin-dependent kinase (CDK1), cyclin B1, and anti-apoptotic proteins (Bcl-2, Mcl-1). In addition, luteolin increased the expression of pro-apoptotic proteins (Bid, Bak, Bax, Bad) and activated caspase-3, with a concomitant increased in the levels of cleaved poly-ADP-ribose polymerase (PARP). Moreover, luteolin reduced the cell migration of GBM 8401 cells. Combination treatment of luteolin with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), an anticancer drug used in clinical GBM treatment, enhanced significantly the cytotoxic effect of BCNU in GBM 8401 cells. These results suggest that luteolin may be a potential candidate for GBM adjuvant therapy.
Typhonium flagelliforme is a prominent plant candidate from aroid family, endowing various curative properties against a variety of illness and infections. This tropical plant found in damp, shady habitats and population of south east asian countries used it as alternative curative health supplement. Traditionally, this plant is used as a alternative remedy for cancer. Also, antibacterial and antioxidant activities are well established. This plant has shown promising results as a cough suppressant, which can be helpful in various respiratory tract problems. This review focuses on various biological activities of Typhonium flagelliforme.
Zerumbone (ZER) is a potential anticancer natural compound, isolated from Zingiber zerumbet Smith. In this investigation, the anticancer properties of ZER were evaluated on cancer cells of T-acute lymphoblastic leukemia, CEM-ss. The results showed that ZER has cytotoxic effect against CEM-ss cells with an IC(50) of 8.4 ± 1.9 μg/ml (coefficient of variation < 30%). Comparatively, 5-fluorouracil (positive control), imposed an inhibitory effect on CEM-ss cells with an IC(50) of 1.94 ± 0.06 μg/ml. Scanning electron microscopy (SEM) results revealed abnormalities such as membrane blebbing, holes and cytoplasmic extrusions, all of which are characteristics of apoptosis. In addition, ZER has increased the number of TUNEL-positive stain and the cellular level of caspase-3, the hallmarks of apoptosis, on treated CEM-ss cells. It could be concluded that, ZER was able to produce apoptosis on T-acute lymphoblastic leukemia, CEM-ss. The current findings suggest that ZER might be helpful for improving the usefulness of anticancer agents in the therapy of leukemia.
Herbs have been utilized to treat acute and chronic disorders for thousands of years. The scientific basis for such utilization, however, has been poorly understood. Nonetheless, in the past few decades, significant advances in experimental methodology and molecular biology enabled researchers to investigate the potential use of phytochemicals to treat or manage a plethora of chronic diseases including various types of cancer, inflammatory diseases and cardiovascular abnormalities. This review summarizes some of the molecular mechanisms through which several phytochemicals may inhibit inflammation and cancer. In addition, the review discusses basic definitions, ethical and clinical aspects and some guidelines for the use of phytochemicals as chemopreventive or therapeutic agents.
Aqueous extracts prepared from six South African medicinal plants, with cancer-related ethnobotanical uses, were tested for their cytotoxic ability in vitro against three human cancer cell lines: DU-145 prostate cancer cells, MDA-MB-231 and MCF-7 breast cancer cells and a non-malignant breast cell line, MCF-12A. The plants studied were: Bidens pilosa, Centella asiatica, Cnicus benedictus, Dicoma capensis, Hypoxis hemerocallidea and Sutherlandia frutescens. Of these plants, only D. capensis exhibited pronounced cytotoxic effects in two of the cell lines tested: MCF-7 and MCF-12A.
Human immunodeficiency virus (HIV) infection is arguably the most significant global health problem of the modern era. Infection
has all but devastated third-world countries and continues to threaten public health in developed nations. With the numbers
of deaths approaching tens of millions each year, the greatest imperative for world health is the realization of an effective
preventative vaccine. The major obstacles prohibiting this goal include a better understanding of protective immunity in the
natural host of the virus. In working toward this objective, animal model systems were developed to recapitulate disease processes
and viral diversity as it occurs in natural infections of man. Nonetheless, HIV is species specific and is diffi- cult to
study in animal systems. Transgenic animals have been developed expressing human receptors in order to overcome some of these
limitations; but an animal model that can be progressively infected by HIV remains elusive. Thus, a number of animal models
have been established that utilize “other” lentiviruses that mimic HIV infection in specific ways and provide the means to
mirror natural infection in its human host. Alternatively, relevant animal models replicate aspects of human disease through
the engraftment of infected human cells. Ultimately, these animal model systems of HIV disease provide insight into specific
disease processes and serve to elucidate underlying mechanisms of infection and subsequent disease.
The MTT cell viability assay is widely used in determining drug sensitivity profiles for patients with hematological malignancies and in primary screening of potential chemotherapeutic drugs. Because the multidrug resistance (MDR) phenotype is associated with these malignancies, and since many vital dyes are effluxed from MDR expressing cells, we have investigated whether the MDR phenotype interferes with the MTT assay. In CCRF-CEM and K562 human leukemic cell lines and drug-resistant sub-lines developed from them, comparison of the MTT assay with other cell viability assays showed significant variation in ic50 concentrations, although the resistance relative to the sensitive parent cell was correlated. Inclusion of verapamil, an inhibitor of drug efflux activity, had no effect an the MTT assay.
yphonium flagelliforme (Lodd.) Blume, commonly known as rodent tuber in Malaysia, is one of the widely used alternative medicines in cancer therapy by South East Asian population. Intake of this plant is common among patients with malignancies especially Leukaemia, breast and cervical cancer; however no data available regarding the possible direct effect of T. flagelliforme in these cancers. The purpose of the present study was to investigate the potential in vitro cytotoxic effect of leaves and tubers of T. flagelliforme extracts against human T4-lymphoblastoid cell line CEM-ss. Among the 8 extracts Dichloromethane and Ethyl acetate extracts of T. flagelliforme demonstrated significant anti proliferative effect with a marked level for both leaves (10.8 and 5.8 μg mL-1) and tuber (6.5 and 8.2 μg mL-1), against CEM-ss cells. Considering all the results collectively T. flagelliforme appears to be a promising plant demonstrating anti cancer activity, that requires further investigation.
This work presents a study of the importance of natural products, especially those derived from higher plants, in terms of drug development. It describes the main strategies for obtaining drugs from natural sources, fields of knowledge involved, difficulties and perspectives. It also includes a brief discussion of the specific situation in Brazil regarding the use of, trade in, and research into therapeutic resources of natural origin and the general lack of awareness of the use of potentially toxic plants, mainly in folk medicine.
Natural product compounds are the source of numerous therapeutic agents. Recent progress to discover drugs from natural product sources has resulted in compounds that are being developed to treat cancer, resistant bacteria and viruses and immunosuppressive disorders. Many of these compounds were discovered by applying recent advances in understanding the genetics of secondary metabolism in actinomycetes, exploring the marine environment and applying new screening technologies. In many instances, the discovery of a novel natural product serves as a tool to better understand targets and pathways in the disease process. This review describes recent progress in drug discovery from natural sources including several examples of compounds that inhibit novel drug targets.
The maximum tolerated dose (MTD) is commonly estimated to be the maximum dose that can be administered for the duration of a specific study that will not compromise the survival of the animals by causes other than carcinogenicity. If the MTD has been chosen appropriately, there should be no adverse effect on survival, only a modest decrement in body weight gain and minimal overt signs of toxicity. The MTD has been exceeded if there is increased mortality, severe body weight decrement, or marked signs of toxicity. It should be noted that another meaning for MTD has sometimes been ‘minimum toxic dose.’
Anin vitro cytotoxicity screening of theTyphonium flagelliforme extracts indicated high cytotoxicity effect on human lung carcinoma NCl-H23 cells and human mammary gland carcinoma T-47D
cells, but the extracts were not active on human liver carcinoma HepG2 cells. NCl-H23 cells were more susceptible toT. flagelliforme extracts than T-47D cells. EDP50 values of the hexane fractions of the mature plant and thein vitro plantlet ofT. flagelliforme on NCl-H23 cells were less than 2 μg/mL Extract from the mature plant was relatively more cytotoxic than the one fromin vitro plantlet except for the hexane fraction. The chloroform and butanol fraction of the mature plant had higher cytotoxicity
effect than the fraction fromin vitro plantlet on NCl-H23 cells. All the 3 fractions (hexane, chloroform, and butanol) of the mature plant exhibited higher cytotoxicity
effects on human mammary gland carcinoma T-47D cells than the 3 fractions ofin vitro plantlet. However, the human liver carcinoma cells were resistant toT. flagelliforme extracts except for higher concentration of hexane fractions of both the mature and thein vitro plants and the chloroform fraction of the mature plant. Micropropagated plantlets ofT. flagelliforme could hence be used as herbal materials for the treatment of human lung and breast cancers.
In recent years, large pharmaceutical companies have significantly reduced or eliminated the search for new therapeutic agents from natural sources. In spite of the many successes from natural product drug discovery, these companies have chosen to focus on compound libraries as the source of new lead compounds. Smaller biotechnology companies are continuing the search for novel natural products by developing and employing new and innovative approaches. This paper will describe some of these recent approaches to natural product drug discovery.:
The antiproliferative effect of an immature Citrus grandis Osbeck fruit extract was investigated using U937 human leukaemia cells. Maximum cytotoxicity was observed using the hexane fraction (HF) of the extract. Cell death was dose-dependent (IC50=ca. 60μg/ml) and was characterised by chromatin condensation, apoptotic body formation, and DNA fragmentation. The induction of apoptosis was confirmed by caspase-3 activity assays and by immunoblotting using antibodies against Bcl-2, Bax, poly(ADP-ribose) polymerase (PARP), caspase-9, and caspase-3. The molecular mechanism underlying HF-induced apoptosis in U937 cells may involve a mitochondria-mediated signalling pathway, as demonstrated by an increase in the Bax/Bcl-2 expression ratio. Analyses of the HF by gas chromatography (GC) and GC-mass spectrometry (MS) tentatively identified 19 compounds, including γ-sitosterol (17.5%), 7-methoxy-8-(2-oxo-3-methylbutyl) coumarin (6.8%), stigmasterol (3.8%), and campesterol (3.4%). Together, our results provide the first evidence that the HF of an immature C. grandis Osbeck fruit extract induces apoptosis in U937 cells.
It is well documented that enhanced garlic (Allium sativum) consumption leads to decrease in the cancer incidences. Diallyl sulfide (DAS), one of the components of garlic, induces cytotoxicity and apoptosis in many cancer cell lines. The present studies are focused on the in vivo effects of DAS on leukemia WEHI-3 cells in the BALB/c mice. We examined the effects of DAS on the cytotoxicity of WEHI-3 cells and results indicated that DAS decreased the percentage of viable WEHI-3 cells and these effects are dose-dependent. We examined the effects of DAS on WEHI-3 in vivo and the results indicated that DAS decreased the percentage of Mac-3 and CD11b, indicating that the differentiation of the precursor of macrophage cells was inhibited. DAS stimulated the percentage of CD3 and CD19, indicating that the differentiation of the precursor of T and B cells promoted. The weights of liver and spleen indicated that DAS decreased the weight of these organs after being compared to the control groups. One of the major characteristic of WEHI-3 leukemia is the enlarged spleen in murine after intraperitoneal (i.p.) injection of WEHI-3 cells. In conclusion, DAS affects WEHI-3 cells both in vitro and in vivo.
Typhonium flagelliforme is an indigenous plant of Malaysia and is used by the local communities to treat cancer. This study aims to identify the chemical constituents of Typhonium flagelliforme particularly those which have antiproliferative properties towards human cancer cell lines.
Purification of the chemical constituents by various chromatographic procedures was guided by the antiproliferative activity. Identification of the chemical constituents was carried out by various spectroscopic techniques including high resolution MS and NMR. The antiproliferative activity was assayed using MTT on NCI-H23 (lung cancer) and HS578T (breast cancer) cell lines. Microscopic observation and DeadEnd colourimetric TUNEL assay was used to identify the apoptotic mode of cell death.
Four pheophorbide related compounds, namely pheophorbide-a, pheophorbide-a', pyropheophorbide-a and methyl pyropheophorbide-a were identified in the most active fraction, D/F19. These constituents exhibited antiproliferative activity against cancer cells and the activity increased following photoactivation. However, the greater antiproliferative activity exhibited by D/F19 itself compared to the pheophorbides and its other subfractions suggests some form of synergistic action between the constituents. The inhibitory effect of D/F19 and the pheophorbides was apoptotic in the absence of light. Other chemical constituents that have been identified in this study include hexadecanoic acid, oleic acid, linoleic acid, linolenic acid, campesterol, stigmasterol and beta-sitosterol. Most of the chemical constituents identified in this plant have not been reported previously.
Enhanced flavonoid consumption is closely related with a reduced cancer incidence as shown in epidemiological studies. Quercetin (3,5,7,3',4'-pentahydroxylflavone) is one of the active components of flavonoids which exist in natural plants, particularly in onions and fruits. It was reported that quercetin induced apoptosis in human cancer cell lines, including human leukemia HL-60 cells, but there is no available information as to its effects on leukemia cells in vivo. The purpose of the present studies was to focus on the in vivo effects of quercetin on leukemia WEHI-3 cells. The effects of quercetin on WEHI-3 cells injected into BALB/c mice were examined. Quercetin decreased the percentage of Mac-3 and CD11b markers, suggesting that the differentiation of the precursors of macrophages and T cells was inhibited. There was no effect on CD3 levels but increased CD19 levels. Quercetin decreased the weight of the spleen and liver compared with the olive oil treated animals. Quercetin stimulated macrophage phagocytosis of cells isolated from peritoneum. Quercetin also promoted natural killer cell activity. Based on pathological examination, an effect of quercetin was observed in the spleen of mice previously injected with WEHI-3 cells. Apparently, quercetin affects WEHI-3 cells in vivo.
Increasing attention is being paid to the possibility of applying chemopreventive agents for the protection of individuals from cancer risk. The beneficial potential of chemoprotective compounds is usually well documented by extensive experimental data. To assure the desired effect, these compounds are frequently concentrated to produce dietary supplements for human use. The additive and synergistic effects of other food constituents are, however, frequently ignored. Even natural chemopreventive compounds have to be considered as xenobiotics. Thus, as much attention has to be paid to their testing prior to their wide application as is usual in drug development for human treatment. Unfortunately, much of the research in this area is solely based on simplified in vitro systems that cannot take into account the complexity of biotransformation processes, e.g. chemopreventive compound-drug interaction, effect on metabolism of endogenic compounds. Hence, the predicted chemopreventive potential is not attained in respect of cancer prevention; moreover, the administration of high doses of chemopreventive compounds might be even detrimental for the human health.
Many evidences have shown that dietary intake of cruciferous vegetables could protect against the risk of various types of malignancies. Benzyl isothiocyanate (BITC), one of the compounds from cruciferous vegetables, had shown induced cell cycle arrest and apoptosis in cancer cells. However, there is no available information to address that BITC affects murine leukemia cells in vitro and in vivo. Here, we investigated in vitro effects of BITC on murine leukemia WEHI-3 cells. BITC decreased the percentage of viable cells via G0/G1 arrest and apoptosis in WEHI-3 cells. BITC induced apoptosis through the dysfunction of mitochondria (decreased the levels of mitochondria membrane potential) and activation of caspase-3. Then we investigated in vivo effects of BITC on murine leukemia WEHI-3 cells and the results indicated that BITC decreased the weights of liver and spleen and it also decreased the percentage of CD11b and Mac-3 markers, indicating that the differentiation of the precursor of macrophage and B cells was inhibited. BITC promoted the activity of macrophage phagocytosis in cells which are isolated from PBMC and peritoneal (i.p.). Taken together, BITC can affect WEHI-3 cells in vitro and in vivo.
Flavonoids are polyphenolic compounds found in various foods of plants. Rutin, one of the flavonoids, had been showed induced apoptosis in cancer cells. There is no available information to address rutin affects murine leukemia cells in vivo. In the present study, we are focused on the in vivo effects of rutin on leukemia WEHI-3 cells. The effects of rutin on WEHI-3 in BALB/c mice in vivo were also examined and the results indicated that rutin decreased the percentage of Mac-3 marker, indicating that the differentiation of the precursor of macrophage and T cells was inhibited. The weights of liver and spleen were decreased from rutin-treated groups compared to the control groups and the results indicated that rutin decreased the weight of these organs. One of the major characteristic of WEHI-3 leukemia is the enlarged spleen in murine after i.p. injection of WEHI-3 cells. After the pathological examination, the function of rutin was observed in the liver and spleen in the mice previously injected with WEHI-3 cells. Rutin promoted the activity of macrophage phagocytosis in cells which isolated from peritoneal (i.p.). Taken together, rutin can affect WEHI-3 cells in vivo.
The influence of Rauscher Leukemia Virus infection of BALB/c mice on the differentiation of red blood cells has been investigated. Twenty-one days after infection, the total number of red blood cells in the spleen is increased 20 times in contrast to the bone marrow which is largely unaffected. The cellular composition of these hemopoietic organs has been analysed using a number of markers of erythroid maturation, i.e. heme and hemoglobin synthesis and the presence of globin messenger RNAs.These studies show that most of the blasts present in leukemic spleens are immature as judged by morphological criteria and do not contain globin mRNA, whereas the more mature blasts contain globin mRNA as determined autoradiographically by in situ hybridization to globin cDNA. The number of mature blasts, comprising about one-third of the immature ones, is both in absolute terms and per mg tissue higher than in control spleens.Pulse labelling with 59Fe shows, that heme and hemoglobin synthesis takes place in leukemic spleens.The rate of heme synthesis in leukemic spleens, as judged by short-term incorporation of 59Fe and 14C-glycine is intermediate between the values for spleens from anemic and control animals.After a pulse, labelled 59Fe is first bound to non-hemoglobin proteins. After a chase, this protein-bound 59Fe is transferred to hemoglobin as erythroid differentiation occurs in spleens of normal and anemic mice. However, this transfer of 59Fe to hemoglobin does not occur in leukemic spleen cells during a pulse/chase in vivo.These results could mean that these immature blasts die or that the maturation is inhibited in the enlarged spleens, whereas only a minor portion differentiates in smaller basophilic erythroblasts, which mature into reticulocytes.
Retinoic acid (RA) has been shown to be an inducer of the terminal differentiation of several leukemia cell lines in vitro and in clinical trials to produce a high percentage of remissions in patients with acute promyelocytic leukemia. In an effort to increase the therapeutic efficacy of RA, we have measured the capacity of granulocyte colony-stimulating factor (G-CSF) to enhance the differentiation inducing activity of RA in WEHI-3B D+ monomyelocytic leukemia cells. Combinations of G-CSF and RA produced a supra-additive increase in the percentage of WEHI-3B D+ cells reducing nitro blue tetrazolium and expressing Mac-1 (CD11b) antigen on the cell surface, two markers of the mature state. In the presence of 50 ng/ml of G-CSF, which produced only 12% differentiation when used alone, 0.5 microM RA induced the same degree of cellular differentiation as the optimum concentration of RA (i.e. 7 microM) employed alone. The supra-additive differentiation produced by this combination was prevented by the presence of G-CSF monoclonal antibody in the culture medium, resulting in a degree of maturation comparable to that produced by the retinoid alone. When cells were sequentially exposed to G-CSF followed by RA, a much higher level of differentiation was obtained than when the order was reversed, suggesting that WEHI-3B D+ cells were primed to enter a differentiation pathway by G-CSF. The supra-additive terminal differentiation exhibited by the mixture of G-CSF and RA suggests that these agents should be evaluated for the therapeutic efficacy of the combination in patients with acute non-lymphocytic leukemia.
Myelomonocytic leukemia was detected in a BALB/c mouse which had undergone mineral oil (paraffin) injections intended to induce plasma cell tumor development. The tumor was composed of a mixed population of monocytic and granulocytic cells. On transplantation of the tumor, four distinct sublines developed, two of which retained the original chloroma appearance and were distinguishable by karyotype (one diploid and one tetraploid). The other two nonchloroma sublines were also distinguishable karyologically, because one had a hypodiploid 39 stemline. Chromosome marker studies in vivo and DNA-content studies of cells from mice carrying the tetraploid subline confirmed that in this leukemia both the monocytic and granulocytic cells are neoplastic, indicating the existence of a neoplastic stem cell capable of differentiation into both cell series. Serum and urine samples from mice carrying this tumor contained high levels (frequently over 200 μg/ml) of muramidase, and cell suspensions of the solid tumor also contained this enzyme. This tumor therefore fulfills all the criteria applied to human myelomonocytic leukemia and is a useful laboratory model for this type of leukemia.
The relationship between maximum tolerated dose (MTD) and study sensitivity for detecting rodent carcinogenicity was evaluated for 216 chemicals found to be carcinogens in laboratory animal studies conducted by the National Cancer Institute (NCI) and the National Toxicology Program (NTP). Approximately two-thirds of these rodent carcinogens would have been detected even without the top dose (estimated MTD), but in many of these studies, some site-specific carcinogenic effects would have been missed. Among the remaining one-third of the rodent carcinogens that required the top dose for statistical significance, approximately 80% had numerically elevated rates of the same site-specific tumors at lower doses as well. Only 13 of the NCI/NTP rodent carcinogens had increased tumor rates limited to the top dose for all sites of carcinogenicity. Alternatively, of the 838 site-specific carcinogenic effects observed in the NCI/NTP studies, 447 (53%) would have been detected even without the top dose. Of the remaining effects, 75% (294/391) showed numerically elevated site-specific tumor rates at lower doses. Our evaluation indicates that most carcinogenic effects observed at the top dose in rodent studies are also present (with reduced incidence that might or might not be statistically significant) at the lower doses typically employed (1/2MTD, 1/4MTD).
To evaluate the effects of a novel 2-aminosteroid, 2-(4'-methyl-1'-piperazinyl)-3alpha-hydroxyl-5alpha-androstane-17-one (KH), on in vitro murine WEHI-3B leukemia cells, semisolid colony culture, MTT assay, morphological examination, NBT reduction, NSAE test and ACP assay were used to determine proliferation and differentiation. It was found that the growth of leukemia cells in colony and liquid cultures was inhibited by KH (10(-8)-10(-4) mol/l) after treatment for 7 days. The percentages of NBT and NSAE positive cells were 71.17 and 79.25%, respectively, after treatment with KH (10(-8)-10(-6) mol/l) for 5 days. The morphology of treated leukemia cells was identified to be macrophage-like and these cells acquired significant ACP activities. It was indicated that the ACP enzyme activities were increased as high as two and three times of the control, respectively, after treatment with 10(-8) or 10(-5) mol/l KH for 6 days. It was also indicated by DNA fragmentation in gel electrophoresis that WEHI-3B cells were induced toward apoptosis by KH (10(-8)-10(-4) mol/l) when checked at day 5. The c-myc mRNA expressions in WEHI-3B cells were decreased by 58.7% after treatment with KH (10(-8) mol/l) for 5 days. Therefore, it is first reported here that KH, a novel 2-aminosteroid, could suppress proliferation and induce differentiation of WEHI-3B leukemia cells. These differentiated cells were mature macrophage-like cells and showed characteristics of functional phagocytes acquired with acid phosphatase activity. The mechanisms underlying the above effects involved the apoptosis of WEHI-3B leukemia cells and the down-regulation of c-myc oncogene expression. It is also shown that the counts of immature granulocytes and monocytes were significantly decreased in both peripheral blood and bone marrow of BALB/c leukemia mice after KH was administrated per os for 7 consecutive days with four doses (5, 10, 15 or 20 mg/kg day), respectively. It is also observed that the enlarged spleens in leukemia mice were decreased when compared with the control.
The plant Typhonium flagelliforme (Araceae), commonly known as the 'rodent tuber', is often included as an essential ingredient in various herbal remedies recommended for cancer therapies in Malaysia. Various extracts prepared from either the roots, tubers, stems or leaves were tested for cytotoxic activity on murine P388 leukaemia cells using the MTT assay method. Both the chloroform (IC50 = 6.0 microg/mL) and hexane (IC50 = 15.0 microg/mL) extract from the 'roots and tubers' exhibited weak cytotoxic activity. The hexane extract (IC50 = 65.0 microg/mL) from the 'stems and leaves' exhibited weaker cytotoxic activity than the chloroform extract (IC50 = 8.0 microg/mL). Although the juice extract from the 'roots and tubers' is frequently consumed for cancer treatment, it exhibited poor cytotoxic activity. Further analysis using an amino acid analyser revealed that the juice extract contained a high concentration of arginine (0.874%). A high tryptophan content (0.800%) was confirmed by NMR and HPLC analysis.
The problem of cancer in Malaysia is a growing one. It is now the fourth leading cause of death among medically certified deaths. Cancer of the lung is the most common killer among malignancies. It is estimated that the annual incidence of cancer is 30 000. The majority of patients are found at a late stage of the disease. The National Cancer Control Program aims to reduce the incidence and mortality of cancer and to improve the quality of life of cancer patients. Policies encompass prevention, early diagnosis, treatment, palliative care and rehabilitation. The program for prevention includes an anti-smoking campaign and immunization of babies against hepatitis B. Papanicolaou's smear and breast self-examination are among efforts for the early detection of cancer. Public education and the promotion of healthy lifestyles have been actively carried out. Facilities for treatment and palliative care are being developed further. Networks between the public and private sectors and non-governmental organizations have been on-going. Apart from the establishment and upgrading of treatment facilities, the need for training of skilled staff in the treatment of cancer is highlighted.
We studied the effects of the in vivo administration of interleukin-2 (IL-2) (at low doses) + GM-CSF in BALB/C mice injected intraperitoneally with LSTRA murine leukemic cell line in order to test the possible role of this immunotherapeutic approach in the eradication of leukemia. Mice were injected intraperitoneally on day -1 with different concentrations of LSTRA cells. On day 0, mice received a lethal dose of TBI of 700 cGy from a Cs(137) source, followed by a single high dose of recombinant human-granulocyte-colony stimulating factor (rh-G-CSF) (1 microg/g) intraperitoneally 2h after TBI. This procedure rescued 80% of mice but only mice injected with the lower concentration of LSTRA cells (10(3)) could be cured. The lethal dose (LD 100/60) of LSTRA in normal mice was 10(4). The subcutaneous administration of rh-IL-2 (4.000 IU/mouse per day, from day +1 to +28) + rm-GM-CSF (1 microg/kg per day) from day +7 to +28 cured mice injected with 10(4) LSTRA cells and 40% of mice injected with 10(5) cells. Mice injected with 10(6) cells died from leukemia. We observed an increase in LAK activity on days +21 and +28 without an increase in NK activity. We show that BALB/C mice injected with LSTRA cell line can be cured by in vivo activation with IL-2 + GM-CSF depending on the cell dose injected. The curability of leukemia was confirmed at the molecular level with a PCR method.
The indolequinone compound EO9 has good pharmacodynamic properties in terms of bioreductive activation and selectivity for either NAD(P)H:quinone oxidoreductase-1 (NQO1)-rich aerobic or NQO1-deficient hypoxic cells. However, its pharmacokinetic properties are poor and this fact is believed to be a major reason for EO9's lack of clinical efficacy. The purpose of this study was to develop quinone-based bioreductive drugs that retained EO9's good properties, in terms of bioreductive activation, but have improved pharmacokinetic properties. Out of 11 naphthoquinone compounds evaluated, 2-aziridinyl-5-hydroxy-1,4-naphthoquinone (compound 2), 2,3-bis(aziridinyl)-5-hydroxy-1,4-naphthoquinone (compound 3), and 2-aziridinyl-6-hydroxymethyl-1,4-naphthoquinone (compound 11) were selected for further evaluation based on good substrate specificity for NQO1 and selectivity towards NQO1-rich cells in vitro. Compound 3 was of particular interest as it also demonstrated selectivity for NQO1-rich cells under hypoxic conditions. Compound 3 was not metabolised by murine whole blood in vitro (in contrast to compounds 2, 11 and EO9) and pharmacokinetic studies in non-tumour-bearing mice in vivo (at the maximum soluble dose of 60 mg kg(-1) administered intraperitoneally) demonstrated significant improvements in plasma half-life (16.2 min) and AUC values (22.5 microM h) compared to EO9 (T(1/2) = 1.8 min, AUC = 0.184 microM h). Compound 3 also demonstrated significant anti-tumour activity against H460 and HCT-116 human tumour xenografts in vivo, whereas EO9 was inactive against these tumours. In conclusion, compound 3 is a promising lead compound that may target both aerobic and hypoxic fractions of NQO1-rich tumours and further studies to elucidate its mechanism of action and improve solubility are warranted.
Arsenic trioxide has shown great promise in the treatment of patients with relapsed or refractory acute promyelocytic leukemia (APL). In clinical trials, arsenic trioxide induces complete remission in 87% of patients and molecular remission in 83% of patients. Two-year overall and relapse-free survival estimates are 63% and 49%, respectively. Treatment with arsenic trioxide may be associated with the APL differentiation syndrome, leukocytosis, and electrocardiographic abnormalities. The expanded use of arsenic trioxide in APL for postremission therapy, in conjunction with transplantation, and in patients with newly diagnosed APL is under investigation. The multiple mechanisms of action of arsenic trioxide suggest that it may have antitumor activity in malignancies other than APL and that it may be used in combination with other agents to expand its potential use. This article reviews the clinical use of arsenic trioxide to date and discusses new therapeutic strategies evolving from its diverse biologic activities.
Podocytes are terminally differentiated and highly specialized epithelial cells. The factors governing podocyte differentiation are poorly understood. We tested the hypothesis that all-trans retinoic acid (ATRA), a vitamin A derivative, induces podocyte differentiation in vitro and in vivo.
We tested the effects of ATRA on podocytes. Primary rat, primary mouse, and immortalized mouse podocytes were exposed to ATRA (1, 5, 10, 20, 40, 50, 80, 160, and 200 micromol/L) or control (ethanol) for 72 hours. Cell morphology was examined by electron microscopy, the expression of podocyte specific proteins was measured by immunoflourescence and Western blot analysis, cell number and apoptosis were measured by 3-[4,5] dimethylthiazol-2,5-diphenyltetrazolium bromide (MTT) assay and Hoechst staining, respectively. To determine if ATRA alters podocyte differentiation in vivo, experimental injury was induced in C57BL6 mice using the antiglomerular antibody. Animals were given either daily intraperitoneal ATRA (16 mg/kg) or vehicle (corn oil). For end points, we measured proteinuria, podocyte-specific protein immunostaining, and proliferation [proliferating cell nuclear antigen (PCNA)] at days 5 and 14 (N= 5/group/time point).
ATRA induced podocyte process formation in vitro, and significantly increased the expression of nephrin and podocin. This coincided with a reduction in proliferation. ATRA also significantly prevented the decrease in staining for synaptopodin, nephrin, and podocin in experimental animals (P < 0.05 vs. control). This was accompanied by reduced proteinuria and decreased podocyte proliferation (P < 0.05 vs. control).
ATRA induces podocyte differentiation in vitro and in vivo and alters the expression of certain podocyte-specific proteins. Further studies are ongoing to delineate the mechanism of this effect.
During the past 15 years, most large pharmaceutical companies have decreased the screening of natural products for drug discovery in favor of synthetic compound libraries. Main reasons for this include the incompatibility of natural product libraries with high-throughput screening and the marginal improvement in core technologies for natural product screening in the late 1980s and early 1990 s. Recently, the development of new technologies has revolutionized the screening of natural products. Applying these technologies compensates for the inherent limitations of natural products and offers a unique opportunity to re-establish natural products as a major source for drug discovery. Examples of these new advances and technologies are described in this review.
This study was designed to identify the cell surface protein markers that can differentiate between chronic myeloid leukemia (CML) and acute promyelocytic leukemia cells (APL). The differentially expressed plasma membrane proteins were analyzed between CML cell line (K562) and APL cell line (NB4) using the comparative proteomic approach. The cell membrane proteins were enriched by labeling with a membrane-impermeable biotinylation reagent, sulfo-NHS-SS-Biotin, and subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS). By comparative proteomic analysis of K562 and NB4 cells, we identified 25 membrane and 14 membrane-associated proteins. The result of LC-MS/MS combined with chemical tagging method was validated by confirming the expression and localization of one of the differentially expressed plasma membrane proteins, CD43, by FACS and confocal microscopy. Our results indicate that CD43 could be a potential candidate for differentiating CML from APL.
To evaluate the effects of YC-1 on leukemia cell lines, PI incorporation was used to determine cell viability. YC-1 induced a dose- and time-dependent decrease in viability and apoptosis in YC-1-treated U937 cells. YC-1-induced apoptosis is a cyclic guanosine monophosphate (cGMP)-independent pathway. Proteomic analysis showed that the altered proteins include the significant regulation of HSP70, chaperonin, ATP synthase beta chains, and Chain F. Western blotting and immuno-cytochemistry stain showed that YC-1 treatment caused a time-dependent increase in cytosolic Cytochrome c, pro-caspase-9, Apaf-1, and the activation of caspase-9 and -3. Importantly, the in vivo antileukemia effects of YC-1 were evaluated in BALB/c mice inoculated with WEHI-3B orthotopic model. YC-1 enhanced survival rate and prevented the body weight loss in leukemia mice. The enlargement of spleen and lymph nodes were reduced in YC-1 treated than that in leukemia mice. H-E stain of spleen sections revealed that infiltration of immature myeloblastic cells into red pulp was reduced in YC-1-treated group. The apoptotic cells of splenocyte were significantly increased in YC-1 treated than that in leukemia mice by Tdt-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay. Taken together, we conclude that YC-1 acted against U937 cells in vitro via a mitochondrial-dependent apoptosis pathway, and in orthotopic leukemia model, YC-1 administered antileukemia activity.
Typhonium flagelliforme (Lodd.) Blume (Araceae) is a Malaysian plant used locally to combat cancer. In order to evaluate its antiproliferative activity in vitro and to possibly identify the active chemical constituents, a bioactivity guided study was conducted on the extracts of this plant.
The active extracts of Typhonium flagelliforme were fractionated by flash column chromatography and each fraction was evaluated for antiproliferative activity using MTT assay. The apoptotic effect of the active fraction was determined microscopically and by using TUNEL colorimetric assay. GC-MS and NMR were used to determine the chemical constituents of this active fraction.
Several fractions of the hexane and dichloromethane extracts were found to inhibit the growth of NCI-H23 non-small cell lung carcinoma cell line significantly, with IC(50)<15 microg/ml. However, most of these active fractions were also found to inhibit the growth of non-tumorigenic BALB/c 3T3 mouse fibroblast cell line except for fraction 21 of the dichloromethane extract (D/F21). This particular fraction was not only less cytotoxic to the non-tumorigenic cells, where the IC(50) was 48.6 microg/ml compared to IC(50) 7.5 microg/ml for NCI-H23, but it was also found to induce apoptosis in the cancer cell line. GC-MS analysis revealed that D/F21 contains hexadecanoic acid, 1-hexadecene, phytol and a derivative of phytol. The presence of non-saturated fatty acids in this fraction was confirmed by nuclear magnetic resonance spectroscopy.
D/F21 was found to be the active and cancer cell line specific fraction of Typhonium flagelliforme. Its major chemical constituents had been determined spectroscopically.
Animal model systems of HIV-diseases. In vivo models of HIV disease and control. Springer publication
- Anderson Er H Xiong
- Gendelman
- He
Anderson ER, Xiong H, Gendelman HE. Animal model systems of HIV-diseases. In vivo models of HIV disease and control. Springer publication; 2005. pp. 19–25.
Maximum Tolerated Dose (MTD). Encyclopedia of Toxicology
- Gad
- Sc
- Philip
Gad SC, Philip W. Maximum Tolerated Dose (MTD). Encyclopedia of Toxicology. New York: Elsevier; 2005. pp. 21–2.
Typhonium divaricatum (rodent tuber): a promising local plant in the fight against cancer
- Neoh
Neoh CK. Typhonium divaricatum (rodent tuber): a promising local plant in the
fight against cancer. Med J Malaysia 1992;47:86.
New aspects of natural products in drug discovery
- Lam