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In vitro antitumor activity of Lasota strain. (A) IC 50 of Lasota strain in AMJ13 cell line (B) IC 50 of Lesotho strain in MCF7 cell line (C) % tumor growth inhibition (GI) of Lasotastrain in AMJ13 (D) % tumor growth inhibition (GI) of Lasota strain in MCF7 cell line, indicated (10,000 cells per well) were transmitted in 96-well flat plates and (0.1,0.5,1, 3, 5,10, 15, 20 MOI) LaSota strain was added in triplicates. After 72 h, tumor growth inhibition was measured by the evolution of MTT. For calculation of % Tumor Growth inhibition (GI), the untreated cell was set to zero.
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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...
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... determine the inhibitory effect of LaSota strain in vitro, we inoculated different MOI (0.1, 0.5, 1, 3, 5, 10,15, 20 MOI) of LaSota strain on monolayers of Human breast carcinoma MCF7 and human breast cancer cell line from Iraq patient AMJ13. After 72 hours, the IC 50 and tumor growth inhibition (GI) were determined ( Table 1, Fig. 1). The anti-tumor behavior of LaSota strain showed that the increase in the dose of LaSota strain led to increasing the inhibitory effect of the virus on breast cancer ...
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... light and inverted fluorescent microscopes were utilized to observe the morphological alterations and the ratios of viable, necrotic and apoptotic cells in the population of human breast cancer cells (AMJ13 and MCF7) exposed LaSota strain IC 50 , 10 MOI for 48h compared to untreated human breast cancer cells. Under the light microscope, those unstained breast cancer cells were observed as showed round and shrinking cells, also we noticed increase in cell granulation, in addition to the changes in unstained culture showed a large empty plaque spaces between cells (Figs. 2 and 3 A2-A3) compared with controls that did not show any change for the same duration (Figs. 2 and 3 A1). Whereas under the light microscope, that stained breast cancer cells with H&E showed that many cytopathic changes, which included vacuolation and cell degeneration, and condensation of the nucleus. ...
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... under the light microscope, that stained breast cancer cells with H&E showed that many cytopathic changes, which included vacuolation and cell degeneration, and condensation of the nucleus. In addition to cells detachment (Figs. 2 and 3 B2-B3), compared with the untreated cells (Figs. 2 and 3 B1). Furthermore, inverted fluorescentmicroscopy utilizing PI (propidium iodide) and AO acridine orange were conducted; the selective permeableness property of the entire plasma membrane of viable cells permitted the acridine orange to enter the cell and was impermeable to the propidium iodide. ...
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... nucleus of viable cells was stained with green when noticed under inverted fluorescence microscope whereas the plasma membrane of necrotic cells was no longer intact which allowed propodium iodide to insert and make the cells appear red. The viable cells can be distinguished from apoptotic cells based on morphological deviations which smaller cells in size and fluoresced orange or red in color, increase in green to red shift were noticed inLaSota strain-infected breast cancer cell line (Figs. 2 and 3 C2-C3), after 48hrs in apoptotic cells as compared with untreated control (viable green cell) (Figs. 2 and 3C1) was observed. This green to red shift in fluorescence was more at 10 MOI than IC\ of LaSota strain; however, 10 MOI of LaSota strain induced more effect (apoptosis), compared to IC 50 dose. ...
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... control 0.441±0.036 0.466±0.041 0.430±0.031 NDV, Caspase8 and Caspase9) were significantly expressed when compared to control, not stained cells (positive control) using ANOVA one-way multiple comparison test (Figs. 4, 5 D1 and D3). However, significant (P<0.05) increase in caspase9 activity and ND antibody in ND viruses infected MCF7 and AMJ13breast cancer cells as compared to caspase8 expression. Further, IC50 of caspase8 in AMJ13 cell line shows no significant (P<0.05) in optical density of expression, protein (Figs. 5 D2), when compared with caspase9 were ...
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... straininfected MCF7 and AMJ13 cells were distinguished by morphological varieties, which include rounding, cell shrinking, and separation from the monolayer. Untreated MCF7 and AMJ13 cells appear normal in its gross morphology. The mechanism(s) of anti-tumor activity for NDV is unknown, may be oncolytic activity was at once submit on the MOI (Fig. 1). Fabian et al (2007) exhibited that a virulent Newcastle virus strain is analytic by used an NDV MTH-68/H of attenuated pathogenic strain, which was originally a vaccine strain. This strain demonstrated oncolytic effect both in vitro and in vivo in a clinical experiment (Schirrmacher et al, 2001). Furthermore, Schirrmacher et al (2001) ...
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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
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.
