Figure 2- - uploaded by Ahmed Majeed Al-Shammari
Content may be subject to copyright.
AMN3cell line treated for 48hrs with IC 50 or10 MOI doses of LaSota strain afters; 400x. (A) In cross microscope, unstained cell showing the rounded cell (orange arrow) and vacuole degeneration (yellow arrow). (B) In H &E staining, a treated cell showing vacuole degeneration (yellow arrow), hypertrophy, and condensation of nuclei (red arrow) compared to an untreated control cell. (C) In acridine orange and propidium iodide staining, a treated cell showed green, viable cells (blue arrow) and red apoptosis cells (white arrow) viewed under a fluorescence microscope.
Source publication
Newcastle disease virus (NDV) is a wide-spectrum anti-tumor agent. The oncolytic selectivity of NDV, a family of Paramyxoviridae, depends on the differential type of inducing different death pathways. This work was conducted to further understand the oncolytic effect of LaSota strain. A mouse breast cancer model (Murine mammary adenocarcinoma cell...
Contexts in source publication
Context 1
... addition, the changes in the unstained culture showed large empty plaque spaces between cells (Figure-2 A2-A3) compared with the control cells that did not show any change for the same duration (Figure-2 A1). Whereas under the light microscope, the H&E stained AMN3 cell line showed t many cytopathological changes which included vacuolation, cell degeneration, condensation of nucleus, and cells detachment ( Figure-2 B2, B3), compared with the untreated cells Figure-(2B1). Furthermore, inverted fluorescent microscopy examination utilizing PI and AO was conducted; the selective permeability property of the entire plasma membrane of viable cells permitted the entry of AO stain, whereas they were impermeable to PI staining. ...
Context 2
... viable cells could be distinguished from apoptotic cells based on morphological deviations such as smaller cells size and fluoresced orange or red color. In addition, an increase in green to red shift was noticed in apoptotic LaSota strain-infected AMN3 cells (Figure-2C2-C3) after 48hrs , as compared with untreated control (viable green cell) (Figure-2 C1). This green to red shift in fluorescence was more at the treatment with the 10 MOI doses than IC50 of LaSota strain; however, 10 MOI of LaSota strain induced more effect (apoptosis, compared to the IC 50 dose. ...
Context 3
... assay was performed to identify the mechanism of apoptosis induction and determine the exact in vitro pathway for apoptosis of LaSota-treated AMN3 cell line after 48h post inoculation. Immunocytochemistry analysis showed that all tumor cells infected with LaSota strain were positive for the expression of ND and caspase 8 and 9 proteins Figures-(3A2-3, B2-3 and C2-3). Antigens in the infected cells were stained in brown, while no brown staining was observed in the positive control (uninfected cells) that were not exposed to LaSota strain but exposed to both primary and secondary antibodies Figures-(3A1, B1 and C1) (Nuclei were stained blue by counterstain). ...
Similar publications
Hematological malignancies remain one of the leading causes of death worldwide despite advances in cancer therapeutics. Newcastle disease virus (NDV) is a member of Paramyxoviridae that elicits considerable interest as an anticancer agent because it can replicate up to 10 000 times faster in human cancer cells than in most normal cancer cells. Seve...
Newcastle disease virus (NDV) is a wide-spectrum anti-tumor agent. The oncolytic selectivity of NDV, a family of Paramyxoviridae, depends on the differential type of inducing different death pathways. This work was conducted to further understand the oncolytic effect of LaSota strain. A mouse breast cancer model (Murine mammary adenocarcinoma cell...
Hematological malignancies remain one of the leading causes of death worldwide despite advances in cancer therapeutics. Newcastle disease virus (NDV) is a member of Paramyxoviridae that elicits considerable interest as an anticancer agent because it can replicate up to 10 000 times faster in human cancer cells than in most normal cancer cells. Seve...
Citations
... NDV is a natural virus that specifically targets cancer cells, spares normal cells, triggers adaptive immunity against tumors, and is acknowledged as a crucial element in cancer treatment by researchers [19,20]. In 2020, a study by Hassan et al. represented that the NDV LaSota strain exhibited anti-cancer properties tackling both mammary adenocarcinoma of mouse and breast cancer cells [21]. NDV replication occurs autonomously from host cell DNA replication. ...
Background and aims
Cervical cancer (CC) is a common cancer among women, often treated with Doxorubicin (Doxo). Research is underway to explore the use of oncolytic virus (OV) therapy as a means to improve drug efficacy and enhance the immune system’s tumor-fighting capabilities. Hence, our study purposes to evaluate the therapeutic potential of Newcastle disease virus (NDV) in increasing the antitumor efficacy of Doxo in mouse models of CC.
Methods and materials
TC1 cells were administered to C57BL/6 mice (Female) in a range of 6 to 8 weeks age (n = 40) to induce tumor growth. After tumor development, four treatment groups of mice were formed. Treatment were performed through NDV, Doxo, and a combination of both in three groups of treatment twice in a one-week intervention manner, while the control group treated with PBS. Following the last treatment, half of these mice were subjected to euthanize due to the immune-response assessment, and the other half were followed up till they died naturally in a certain period of time.
Results
Mice that underwent the combined treatment showed significantly improved survival rates and slower tumor progression in comparison with the control group. This combined treatment substantially elevated nitric oxide (NO) and lactate dehydrogenase (LDH) levels in the splenocytes cultures of mice bearing cervical tumors. Furthermore, combination therapy resulted in a notable elevation in TNF-α, IL-12, and IFN-γ, secretion alongside a reduction in the release of TGF-β and IL-4 within the splenocytes in counter with the treatment of just NDV or Doxo.
Conclusion
According to the findings of this study, it seems that utilizing NDV can improve the effectiveness of Doxo in a mouse model of CC, suggesting it can serve as an adjunct therapy alongside chemotherapy.
... According to the results of this study, the inhibitory concentration 50% growth (IC50) for oncolytic RSV-A2-infected TC-1 cells after 72 h post-infection was around 10 MOI. Therefore, this MOI was selected as the optimal dose (RSV-A2 IC50 MOI) for future experiments [26][27][28]. ...
Background
Cervical cancer represents one of the most prevalent cancers among women worldwide, particularly in low- and middle-income nations. Oncolytic viruses (OVs) can infect cancer cells selectively and lethally without harming normal cells. Respiratory syncytial virus (RSV) is an oncolytic virus for anticancer therapy because of its propensity to multiply within tumor cells. This research aimed to assess the in vitro antitumor activities and molecular basis processes of the oncolytic RSV-A2 on the TC-1 cancer cells as a model for HPV‑related cervical cancers.
Methods
Cellular proliferation (MTT) and lactate dehydrogenase (LDH) release assays were used to investigate the catalytic impacts of RSV-A2 by the ELISA method. Real-time PCR and flow cytometry assays were utilized to assess apoptosis, autophagy, intracellular concentrations of reactive oxygen species (ROS), and cell cycle inhibition.
Results
Our MTT and LDH results demonstrated that TC-1 cell viability after oncolytic RSV-A2 treatment was MOI-dependently and altered significantly with increasing RSV-A2 virus multiplicity of infection (MOI). Other findings showed that the RSV-A2 potentially resulted in apoptosis and autophagy induction, caspase-3 activation, ROS generation, and cell cycle inhibition in the TC-1 cell line. Real-time PCR assay revealed that RSV-A2 infection significantly elevated the Bax and decreased the Bcl2 expression.
Conclusions
The results indicated that oncolytic RSV-A2 has cytotoxic and inhibiting effects on HPV-associated cervical cancer cells. Our findings revealed that RSV-A2 is a promising treatment candidate for cervical cancer.
... The attenuated strain of (AMHA1) is a naturally oncolytic NDV with extensive anticancer activity [28]. NDV's replication has been evaluated on a variety of human malignancies [29]. After the cancer cell is infected with NDV, it multiplies swiftly and infects neighboring tumor cells by producing the child virions, which are evident three hours after inoculation and form plaques after two days [30]. ...
Glioblastoma multiforme (GBM) is one of the most life-threatening types of cancer that is difficult to treat. The search for effective yet safe therapy is progressing and non-conventional therapies such as using viruses as a smart and selective agent against cancer are promising. The aim of the study was the presence of a reliable method to use Newcastle disease virus (NDV) as an oncolytic agent against GBM, which attempted to propagate the NDV in laboratory experiments. Ahmed Majeed Hamza Al-Shammari-1 (AMHA1) attenuated strain of NDV was propagated in chicken embryos. The virus's tittered in Vero-slamed cells to determine the infective dose. MTT cell viability assay was used to investigate the killing effects of NDV on Ahmed-Majeed-Glioblastoma-Multiforme-2005 (AMGM5) human glioblastoma cancer cells derived from Iraqi patients. The infected cells' morphology was studied to measure the cytolytic effect of the NDV in cancer cells. Results showed that After 24 to 72 hours of inoculation, all of the chicken embryos died when the AMHA1 Iraqi NDV strain was injected. Cell viability assay showed that the NDV-AMHA1 strain has cytotoxicity at MOI of 0.1, 0.5, and 1 for 72 hours of exposure to cancer cells. The morphological analysis showed that NDV induces cell death in the infected cells with both necrotic and apoptotic features. In conclusion, the study focuses on the propagation of the oncolytic NDV as a biological agent capable of overcoming treatment resistance through infecting and replicating inside cancer.
... 53 In addition, one of the recent studies conducted in Iraq confirmed that the NDV is an antitumor agent because it showed a clear effect in inhibiting the growth of AMN3 (mouse breast cancer cells), and it is also suggested as a live vaccine. 54 The advantages of viral therapy are that the oncolytic vectors have the ability to bind perfectly to the cell, enter the tumor cell by receptor-mediated endocytosis easily, proliferate in cancer cells selectively as opposed to normal cells, and have minimal side effects. Each way of administration has several disadvantages according to dosage, specificity, complexity, efficiency, and feasibility. ...
Cancer is one of the most common diseases around the world and the second leading cause of death after cardiovascular disease. Breast cancer is the most prevalent cancer type among Iraqi women, as it represents the highest percentage of malignant tumors in women until 2018. Therefore, women should be aware of the aggravation of this disease, the importance of the periodic examination for early detection for breast cancer, and following the most appropriate means for the treatment to get recovered and, thus, to reduce mortality. To fight cancer, there is an urgent need to search for new effective anticancer therapies that alter the molecular biology of tumor cells, stimulate the immune system, or specifically deliver chemotherapy factors directly to cancer cells without affecting normal cells and reducing the side effects of treatments. In this context, this paper aimed to highlight the therapeutic approaches used in the current researches of breast cancer treatment. Accumulated evidence showed that medicinal plant extracts, and can serve as anticancer agents. The proposed mechanisms were discussed and presented in this review.
... Viruses from this family are enveloped, nonsegmented, negative-sense RNA viruses that cause the inflammation of the respiratory and gastrointestinal tracts in a wide variety of poultry species [28]. The virulent NDV strain still inducing outbreaks in different countries, including Iraq, leading to severe economic damages in the poultry industry [11]; yet, attenuated and lentogenic NDV has promising antitumor activity and excellent safety in laboratory animals [26]. The Iraqi NDV strain AMHA1 is an oncolytic virus that is attenuated strain isolated originally from an outbreak [7]. ...
Newcastle disease virus (NDV) can modulate cancer cell signaling pathway and induce apoptosis in cancer cells. Cancer cells increase their glycolysis rates to meet the energy demands for their survival and generate ATP as the primary energy source for cell growth and proliferation. Interfering the glycolysis pathway may be a valuable antitumor strategy. This study aimed to assess the effect of NDV on the glycolysis pathway in infected breast cancer cells. Oncolytic NDV attenuated AMHA1 strain was used in this study. AMJ13 and MCF7 breast cancer cell lines and a normal embryonic REF cell line were infected with NDV with different multiplicity of infections (moi) to determine the IC50 of NDV through MTT assay. Crystal violet staining was done to study the morphological changes. NDV apoptosis induction was assessed using AO/ PI assay. NDV interference with the glycolysis pathway was examined through measuring hexokinase (HK) activity , pyruvate, and ATP concentrations, and pH levels in NDV infected and non-infected breast cancer cells and in normal embryonic cells. The results showed that NDV replicates efficiently in cancer cells and spare normal cells and induce morphological changes and apoptosis in breast cancer cells but not in normal cells. NDV infected cancer cells showed decreased in the HK activity, pyruvate and ATP concentrations, and acidity, which reflect a significant decrease in the glycolysis activity of the NDV infected tumor cells. No effects on the normal cells were observed. In conclusion, oncolytic NDV ability to reduce glycolysis pathway activity in cancer cells can be an exciting module to improve antitumor therapeutics.
Background
Breast cancer is one of the most common malignancies in women, with one in 20 globally. Oncolytic viruses have recently been the first step in the biological treatment of cancer, either genetically engineered or naturally occurring. They increase specifically inside cancer cells and destroy them without damaging normal tissues or producing a host immune response against tumour cells or expressing transgenes. One of the most known members of this family is the Newcastle disease virus (NDV), a natural oncolytic virus that selectively induces apoptosis and DNA fragmentation in human cancer cells.
Methods
This study performed biochemical and molecular investigations with variable doses of NDV (32, 64, 128 HAU) and liposomal doxorubicin (9 mg/kg) on mouse triple‐negative mammary carcinoma cell line 4T1 and BALB/c models tumours for the first time.
Results
Real‐time quantitative PCR analysis in NDV‐treated animal tumours showed increased expression of P21, P27 and P53 genes and decreased expression of CD34, integrin Alpha 5, VEGF and VEGF‐R genes. Additional assessments in treated mouse models also showed that NDV increased ROS production, induced apoptosis, reduced tumour size and significantly improved prognosis, with no adverse effect on normal tissues.
Conclusions
These findings all together might indicate that NDV in combination with chemotherapy drugs could improve prognosis in cancer patients although many more conditions should be considered.
Newcastle Disease Virus (NDV) can modulate cancer cell signaling pathways and induce apoptosis in cancer cells. The laboratory-based studies of the oncolytic NDV requires a reliable protocol for the propagation of the oncolytic NDV. A comprehensive protocol is provided for virus propagation in fertile chicken eggs, which consistently yields high titer viral stock. Aim: Propagation of oncolytic NDV AMHA1 attenuated strain in Embryonated Chicken Eggs (ECE) and tissue culture infective dose 50% (TCID 50 ) determination protocol of the virus. Method: Specific pathogen-free fertilized chicken eggs were incubated at 37 °C and 55-60% humidity for 9’ 10 days. Over this period, embryo death was monitored using an egg candle regularly. Virus inoculation is carried out by injection of the diluted virus stock into the allantoic cavity using a needle. embryo death was recorded every two hours and the egg rushed to the refrigerator and fluids collected after four to six hours. Hemagglutination assay (HA) was used to determine the preliminary titer of the virus to collect the high titer egg fluids only which is about 128 to 256HAU. The Vero cell line was exposed to NDV at tenfold serial dilutions to determine TCID 50 of the virus. The number of viruses in 1 ml of allantoic fluid was measured of embryonated chicken eggs. Results: NDV Iraqi virulent strain has the ability to kill all the chicken embryos through (24-72) h of inoculation. A high titer of NDV was achieved from the infected eggs.
Conclusion: Oncolytic NDV propagated in embryonated chicken eggs in high titers as indicated by TCID 50 value.