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In-vitro cytotoxic screening of Glycyrrhiza glabra L. (Fabaceae): A natural anticancer drug

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Abstract

During the last few decades, chemo preventive and chemotherapeutic compounds against various types of cancer have been isolated from various plants. Glycyrrhezic acid (2-20%), Glycyrrhizin, is present as a major chemical constituent of natural anticancer compound Glycyrrhiza glabra L. (Fabaceae). The present work is aimed to investigate the in-vitro cytotoxic screening of standard 18 β-glycyrrhetic acid and also for natural anticancer drug Glycyrrhiza glabra L. (Fabaceae) using three different extracts (chloroform, methanol and water) of the drug through MTT method. Cell viability of previously identified 18 β-glycyrrhetic acid in three different extracts of Glycyrrhiza glabra L. (Fabaceae) was determined by two fold trypan blue method using two different cell lines MCF7-cancerous and Vero-normal cell line and it was quantified through HPTLC method. The results of the HPTLC study indicated that the amount of 18 β-glycyrrhetic acid into three different extracts (chloroform, methanol and water extract) of Glycyrrhiza glabra L. (Fabaceae) was 26.6, 12.5 and 5.6 μg/g, respectively. The percentage viability of two different cell lines was 45.71% for Vero-normal cell line and 78.78% for MCF7-cancerous cell line. IC 50 values of standard 18 β-glycyrrhetic acid was 0.412±0.004 μM and those for the three different extracts (chloroform, methanol and water)of Glycyrrhiza glabra L. (Fabaceae) on MCF7 cell line were 0.4485±0.001, 0.9906±0.001 and 1.288±0.005 μM, respectively. From the above result, it can be said that 18 β-glycyrrhetic acid could be considered as a potential source of natural anticancer component and the percentage of which was higher in the chloroform extract.

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... [32] The utility of cell lines acquired from tumor allows the investigation of tumor cells in a simplified and controlled environment. [33] Common basic steps of in vitro cytotoxic screening include: (a) Isolation of cells, (b) incubation of cells with drugs, (c) assessment measure of cell viability. [33] It is now well-documented that apoptosis or programmed cell death is the key mechanism by which chemotherapeutic agents exert their cytotoxicity. ...
... [33] Common basic steps of in vitro cytotoxic screening include: (a) Isolation of cells, (b) incubation of cells with drugs, (c) assessment measure of cell viability. [33] It is now well-documented that apoptosis or programmed cell death is the key mechanism by which chemotherapeutic agents exert their cytotoxicity. [34] Cytotoxicity has been defined as the cell killing property of a chemical compound independent from the mechanism of death. ...
... MTT works by being converted to a formazan dye only by metabolic active cells. [33,37] MTT proliferation assay was carried out to determine the growth rate of cells. In this study, DOC-loaded gelatin nanoparticles have indicated significant growth inhibition in MCF-7 cell line at a low concentration of IC 50 values. ...
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Background: The goal of the present investigation was to evaluate docetaxel (DOC)-loaded gelatin nanoparticles for cancer therapy. Materials and Methods: DOC-loaded gelatin nanoparticles using ultraviolet-visible spectroscopy, X-ray diffraction, particle size and size distribution, scanning electron microscopy, drug entrapment efficiency, infrared, and in vitro drug release were characterized. The viability of MCF-7 breast cancer (BC) cells was determined by MTT. Cell sensitivity to drugs and growth curves were measured by MTT assay. Changes of cell morphology and ultrastructure were examined by optical and electron microscopy. Results: Solubility, crystallinity, and the crystal properties of an active pharmaceutical ingredient play a critical role in the value chain of pharmaceutical development, manufacturing, and formulation. The rate of drug release for formulation stored at 45°C ± 1°C was increased as compared with the fresh formulation; it might be due to the formation of more pores in the nanoparticles due to evaporation of residual amount of solvent. The in vitro release studies of drug-loaded nanoparticles were conducted at 37 ± 0.5 and 100 rpm using phosphate buffer pH 7.4 (900 ml) in a USP dissolution apparatus under sink condition. DOC-loaded gelatin nanoparticles depleted the viability of MCF human breast cell line. In this study, MCF BC cell line revealed growth inhibition in a dose-dependent manner when treated DOC-loaded gelatin nanoparticles at concentrations ranging from 5 to 100 µg. Conclusion: The DOC-loaded gelatin nanoparticles displayed differential cytotoxicity toward MCF7 cancer cells. These biogenic nanoparticles are biocompatible and found to be good candidates for sustained drug delivery in diseases like cancer.
... The newly synthesized compounds were screened for their cytotoxic activities using NCI (lung cancer cell line), MCF 7 (breast cancer cell line) and HEK293 (normal kidney cell line) cells by the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5diphenyltetrazolium bromide) assay. 24 The screening experiments were conducted at the National Cancer Institute (NCI, Cairo, Egypt) and undertaken with approval from Taibah University, Al-madinah Al-munawarah, Saudi Arabia (approval #3006/434). The cultures were inspected using an inverted microscope to determine the degree of viability. ...
... Cytotoxic assays were performed according to the reported method. 24 First, the cells were incubated at a concentration of 1 Â 10 6 cells/ml in culture medium for 3 h at 37 C and 6.5% CO 2 . After incubation, the cells were seeded at a concentration of 5 Â 10 4 cells/well in 100 ml of culture medium with different concentrations (0.005e100 mM/ml) of the reference drug methotrexate and the synthesized compounds, which were dissolved in 2% DMSO (dimethylsulphoxide) solution into microplates (tissue culture grade, 96 wells, flat bottom) and incubated for 24 h at 37 C and 6.5% CO 2 . ...
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Abstract Objectives The aim of the present research was to synthesise several novel fluorinated quinazoline-sulphonamide derivatives and to evaluate their in vitro cytotoxic activity. Methods Eight compounds were synthesised. The compounds' anticancer activities were determined through the [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide] (MTT) assay using a three-cell-line panel consisting of National Cancer Institute (NCI) lung cancer cells, Michigan Cancer Foundation-7 (MCF-7) breast cancer cells, and Human Embryonic Kidney-293 (HEK-293) normal kidney cell. The values of C log P correlations were determined to interpret the results. Results One compound exhibited significant anticancer activity with low toxicity compared with the methotrexate as the reference drug. The biological screening showed good to moderate anticancer activity for the title compounds compared with the reference drug. The reference drug exhibited an IC50 value of 2.4 μM, whereas compound 9, which was identified as the most active compound, exhibited an IC50 value of 2.51 μM on the NCI cell line. The other compounds showed IC50 values that ranged from 2.89 to 46.34 μM on the three cell lines. The newly synthesized compounds had lower toxicity on the normal cell line than did methotrexate. Conclusions The newly synthesized compounds may provide a valuable template for future design and optimization to produce analogues that act as more active anticancer agents.
... Isobavachalcone (4) was synthesized through Claisen-Schmidt condensation using 4-hydroxybenzaldehyde with 2 ,4 -dihydroxyacetophenone as starting materials (Scheme S2) [32]. Structures of the final products 3 and 4 were confirmed by comparing their spectroscopic data with those reported in the literatures [33,34]. ...
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Biotransformation of four bioactive phenolic constituents from licorice, namely licoisoflavanone (1), glycyrrhisoflavone (2), echinatin (3), and isobavachalcone (4), was performed by the selected fungal strain Aspergillus niger KCCM 60332, leading to the isolation of seventeen metabolites (5–21). Structures of the isolated compounds were determined on the basis of extensive spectroscopic methods, twelve of which (5–7, 10–17 and 19) have been previously undescribed. A series of reactions including hydroxylation, hydrogenation, epoxidation, hydrolysis, reduction, cyclization, and alkylation was observed in the biotransformation process. All compounds were tested for their cytotoxic activities against three different human cancer cell lines including A375P, MCF-7, and HT-29. Compounds 1 and 12 exhibited most considerable cytotoxic activities against all the cell lines investigated, while compounds 2 and 4 were moderately cytotoxic. These findings will contribute to expanding the chemical diversity of phenolic compounds, and compounds 1 and 12 may serve as leads for the development of potential cancer chemopreventive agents.
... The anti-cancerous property Gg is well documented in different cancer cell lines viz. MCF-7 and HT-29 colon cells (Rathi et al., 2009;Jo et al., 2005;Nazmi, et al., 2018& Nourazarian et al., 2016. To rule out any interference of the phytoextract with the anti-cancerous property of DOX, we simultaneously designed an experimental study with breast cancer line (MDA-MB). ...
Article
Ethnopharmacological relevance: Doxorubicin (DOX) is an effective anti-neoplastic drug, however; it has downside effects on cardiac health and other vital organs. The herbal remedies used in day to day life may have a beneficial effect without disturbing the health of the vital organs. Glycyrrhiza glabra L. is a ligneous perennial shrub belonging to Leguminosae/Fabaceae/Papilionaceae family growing in Mediterranean region and Asia and widespread in Turkey, Italy, Spain, Russia, Syria, Iran, China, India and Israel. Commonly known as mulaithi in north India, G. glabra has glycyrrhizin, glycyrrhetic acid, isoliquiritin, isoflavones, etc., which have been reported for several pharmacological activities such as anti-demulcent, anti-ulcer, anti-cancer, anti-inflammatory and anti-diabetic. Aim of the study: The objective of the present study is to investigate the interaction between the molecular factors like PPAR-α/γ and SIRT-1 during cardiac failure arbitrated by DOX under in vitro conditions and role of Glycyrrhiza glabra (Gg) root extract in alleviating these affects. Materials and methods: In the present study, we have examined the DOX induced responses in H9c2 cardiomyocytes and investigated the role of phytochemical Glycyrrhiza glabra in modulating these affects. MTT assay was done to evaluate the cell viability, Reactive Oxygen Species (ROS)/Reactive Nitrogen Species (RNS) levels, mitochondrial ROS, mitochondrial membrane potential was estimated using fluorescent probes. The oxidative stress in terms of protein carbonylation, lipid peroxidation and DNA damage was detected via spectrophotometric methods and immune-fluorescence imaging. The cardiac markers and interaction between SIRT-1 and PPAR-α/γ was measured using Real-Time PCR, Western Blotting and Co-immunoprecipitation based studies. Results: The Glycyrrhiza glabra (Gg) extracts maintained the membrane integrity and improved the lipid homeostasis and stabilized cytoskeletal element actin. Gg phytoextracts attenuated aggravated ROS level, repaired the antioxidant status and consequently, assisted in repairing the DNA damage and mitochondrial function. Further, the expression of hypertrophic markers in the DOX treated cardiomyocytes reconciled the expression factors both at the transcriptional and translational levels after Gg treatment. SIRT-1 mediated pathway and its downstream activator PPAR factors are significant in maintaining the cellular functions. It was observed that the Gg extract allows regaining the nuclear SIRT-1 and PPAR-γ level which was otherwise reduced with DOX treatment in H9c2 cardiomyocytes. The co-immunoprecipitation (Co-IP) documented that SIRT-1 interacts with PPAR-α in the untreated control H9c2 cardiomyocytes whereas DOX treatment interferes and diminishes this interaction however the Gg treatment maintains this interaction. Knocking down SIRT-1 also downregulated expression of PPAR-α and PPAR-γ in DOX treated cells and Gg treatment was able to enhance the expression of PPAR-α and PPAR-γ in SIRT-1 knocked down cardiomyocytes. Conclusions: The antioxidant property of Gg defend the cardiac cells against the DOX induced toxicity via; 1) reducing the oxidative stress, 2) maintaining the mitochondrial functions, 3) regulating lipid homeostasis and cardiac metabolism through SIRT-1 pathway and 4) conserving the cardiac hypertrophy and hence preserving the cardiomyocytes health. Therefore, Gg can be recommended as a healthier supplement with DOX towards cancer therapeutics associated cardiotoxicity.
... Previous studies with Fabaceae species have shown that the Gymnocladus dioicus extract exhibits a cytotoxic effect on HeLa cells in a concentration-dependent manner (Jantova et al., 2001). Bremner et al. (2009) reported that Ononis ramosissima extract exhibits a cytotoxic effect on HeLa cells, while Rathi et al. (2009) demonstrated that Glycyrrhiza glabra extract exhibits a cytotoxic effect on human breast cancer (MCF-7) cells. (Demir et al., 2019b). ...
... There are several reports of biological activities including anti-inflammatory, antimicrobial, cytotoxic, and antioxidant properties and these beneficial activities being attributed to their polyphenolic compounds (Bremner et al., 2009;Stefanović et al., 2015). Although there are some studies reporting the cytotoxic effects of different Dorycnium species in recent years (Jantova et al., 2001;Rathi et al., 2009), there are limited studies on the cytotoxic effect of D. pentaphyllum . Especially, it has been revealed that some natural product extracts can increase apoptosis in cancer cells without harming normal cells in recent years. ...
... The in-vitro cytotoxic screening of standard 18 β-glycyrrhetic acid and also for natural anticancer drug G. glabra using three different extracts (chloroform, methanol and water) of the drug through MTT method disclose that chloroform extract inhibited abnormal cell proliferation more effectively, shows good cytotoxicity against cancerous MCF7 cells (Human Breast cancer) than the other two extracts of G. glabra. [19] 6. ...
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... The trypan blue dye exclusion assay is the most commonly accepted method for the measurement of cell viability. It relies on the alteration in membrane integrity as determined by the uptake of dye by dead cells, thereby giving a direct measure of cell viability [5]. It is now well documented that apoptosis or programmed cell death is the key mechanism by which chemotherapeutic agents exert the cytotoxicity [6]. ...
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The present research was aimed to evaluate the anti-cancer potential of fucoidan extracted from Padina distromatica Hauck in Hare Island, Thoothukudi, Tamil Nadu, India for the inhibitory effect against A-431, A-549, HL-60, HT-29 and MCF-7 cell lines. The viability percentage of the cell lines were carried out by using Trypan blue dye exclusion method and the cytotoxicity of fucoidan of Padina distromatica Hauck on A-431, A-549, HL-60, HT-29 and MCF-7 cell lines were evaluated by MTT assay. The fucoidan from Padina distromatica Hauck has significant cytotoxicity effect on A-549 cell line followed by moderate activity against HL-60 and A-431. However fucoidan was found to have less activity on HT-29 and MCF-7 in the concentration range between 250 to 1000μg/ml of fucoidan by using MTT assay. From the performed assay, it was concluded that fucoidan extracted from Padina distromatica Hauck shows greater activity on all the tested cell lines which can be used as anti-cancer activity.
... All the synthesised compounds were further evaluated for their in vitro breast cancer against MCF-7 cell line using MTT assay as followed by the literature [37,38] with slight modification in dosage for treating cells. The susceptibility of cells to the tested compound aliquots and standard doxorubicin was characterised by IC 50 ( Table 2) and the graph interpretation was done using Graphpad Prism Software (6.01v). ...
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... MTT is also used to verify protective effect as it evaluates cell viability in turns. [24] Considering that 100 alkaloids structures have been isolated and elucidated from Amaryllidaceae family plant, with antiproliferative activity over eukaryotic cells by interfering with protein synthesis, [25] this work proposes to select isolated alkaloids from this family and to evaluate cytotoxicity comparing the traditional colorimetric MTT method with image data generated by CellProfiler®. Isolated alkaloids from the Amaryllidaceae family as licorine, 3-epimacronine, tazettine, trispheridine, albomauline, and galanthine. ...
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... The reported percentage of licorice extract cytotoxic activity was 63% at 0.24 mg/mL with greater percentages apparent as the concentrations of licorice were increased up to 4.8 mg/mL. Rathi et al. (66) reported that IC 50 values of chloroform, methanol, and water extracts of G. glabra L. in the breast MCF7 cell line were 0.45, 0.99, and 1.29 µM, respectively. In the present study, the G. glabra water extract displayed mutagenic activity against the TA100 strain in the presence of metabolic activation, but only at the highest extract concentration. ...
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... The result of the study showed glycyrrhetic acid is a potential source of natural anticancer component and the percentage of which was found higher in the chloroform extract. [19][20] Cannabis sativa: ...
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