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Inhibitory and apoptosis-inducing effects of Newcastle disease virus strain AF2240 on mammary carcinoma cell line.

Ahmad U, Ahmed I, Keong YY, Abd Manan N, Othman F - Biomed Res Int (2015)

Bottom Line: Surgery, radiotherapy, and chemotherapy are the available treatments for breast cancer, but these were reported to have side effects.Morphological and apoptotic-inducing effects of NDV on MD-MB-231 cells were observed using phase contrast and fluorescence microscopes.It appears that NDV AF2240 strain is a potent anticancer agent that induced apoptosis in time-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratory of Anatomy & Histology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia ; Department of Human Anatomy, Faculty of Medicine, Bauchi State University, PMB 65, Gadau, Nigeria.

ABSTRACT
Breast cancer is the malignant tumour that developed from cells of the breast and is the first leading cause of cancer death among women worldwide. Surgery, radiotherapy, and chemotherapy are the available treatments for breast cancer, but these were reported to have side effects. Newcastle disease virus (NDV) known as Avian paramyxovirus type-1 (APMV1) belongs to the genus Avulavirus in a family Paramyxoviridae. NDV is shown to be a promising anticancer agent, killing tumour cells while sparing normal cells unharmed. In this study, the oncolytic and cytotoxic activities of NDV AF2240 strain were evaluated on MDA-MB-231, human mammary carcinoma cell line, using MTT assay, and its inhibitory effects were further studied using proliferation and migration assays. Morphological and apoptotic-inducing effects of NDV on MD-MB-231 cells were observed using phase contrast and fluorescence microscopes. Detection of DNA fragmentation was done following terminal deoxyribonucleotide transferase-mediated Br-dUTP nick end labeling staining (TUNEL) assay, which confirmed that the mode of death was through apoptosis and was quantified by flow cytometry. Furthermore, analysis of cellular DNA content demonstrated that the virus caused an increase in the sub-G1 phase (apoptotic peak) of the cell cycle. It appears that NDV AF2240 strain is a potent anticancer agent that induced apoptosis in time-dependent manner.

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Related in: MedlinePlus

Contour diagram of BrdUTP/PI flow cytometry for MDA-MB-231 breast cancer cell lines treated with NDV AF2240 IC50 concentration value. (a) Untreated cells after (b) 24 hours (c) 48 hours, and (d) 72 hours of treatment. The lower left quadrants of each panel (R1) show the viable cells, which exclude PI and are negative for BrdUTP binding. The upper right quadrants (R2) contain the nonviable, necrotic cells, positive for BrdUTP binding and for PI uptake. The lower right quadrants (R3) represent the fragmented DNA (apoptotic cells), BrdUTP positive, and PI negative.
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fig7: Contour diagram of BrdUTP/PI flow cytometry for MDA-MB-231 breast cancer cell lines treated with NDV AF2240 IC50 concentration value. (a) Untreated cells after (b) 24 hours (c) 48 hours, and (d) 72 hours of treatment. The lower left quadrants of each panel (R1) show the viable cells, which exclude PI and are negative for BrdUTP binding. The upper right quadrants (R2) contain the nonviable, necrotic cells, positive for BrdUTP binding and for PI uptake. The lower right quadrants (R3) represent the fragmented DNA (apoptotic cells), BrdUTP positive, and PI negative.

Mentions: Figure 7 shows the results for flow cytometric analysis of BrdUTP/PI stained MDA-MB-231 cells posttreated with IC50 concentration value of NDV AF2240 strain at 24 (Figure 7(b)), 48 (Figure 7(c)), and 72 hours periods (Figure 7(d)), respectively. The lower left quadrant (R1) of the cytograms is indicating the total number of viable cells, which excluded PI and were negative for BrdUTP binding, while the upper right quadrant (R2) represents the nonviable (necrotic cells) which is positive for BrdUTP binding and showing PI uptake. The lower right quadrant (R3) represents the number of cells that had DNA fragmentation (apoptotic cells) and BrdUTP positive but negative for PI stain, indicating BrdUTP binding. The BrdUTP + PI apoptotic cell population for MDA-MB-231 cell line increased significantly (*P < 0.05) from 1.71 ± 0.02% in untreated cells to 11.66 ± 0.91% in cells treated for 24 hours and to 36.98 ± 2.71% and 44.52 ± 10.13% in cells treated for 48 and 72 hours (Table 1), respectively.


Inhibitory and apoptosis-inducing effects of Newcastle disease virus strain AF2240 on mammary carcinoma cell line.

Ahmad U, Ahmed I, Keong YY, Abd Manan N, Othman F - Biomed Res Int (2015)

Contour diagram of BrdUTP/PI flow cytometry for MDA-MB-231 breast cancer cell lines treated with NDV AF2240 IC50 concentration value. (a) Untreated cells after (b) 24 hours (c) 48 hours, and (d) 72 hours of treatment. The lower left quadrants of each panel (R1) show the viable cells, which exclude PI and are negative for BrdUTP binding. The upper right quadrants (R2) contain the nonviable, necrotic cells, positive for BrdUTP binding and for PI uptake. The lower right quadrants (R3) represent the fragmented DNA (apoptotic cells), BrdUTP positive, and PI negative.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4363544&req=5

fig7: Contour diagram of BrdUTP/PI flow cytometry for MDA-MB-231 breast cancer cell lines treated with NDV AF2240 IC50 concentration value. (a) Untreated cells after (b) 24 hours (c) 48 hours, and (d) 72 hours of treatment. The lower left quadrants of each panel (R1) show the viable cells, which exclude PI and are negative for BrdUTP binding. The upper right quadrants (R2) contain the nonviable, necrotic cells, positive for BrdUTP binding and for PI uptake. The lower right quadrants (R3) represent the fragmented DNA (apoptotic cells), BrdUTP positive, and PI negative.
Mentions: Figure 7 shows the results for flow cytometric analysis of BrdUTP/PI stained MDA-MB-231 cells posttreated with IC50 concentration value of NDV AF2240 strain at 24 (Figure 7(b)), 48 (Figure 7(c)), and 72 hours periods (Figure 7(d)), respectively. The lower left quadrant (R1) of the cytograms is indicating the total number of viable cells, which excluded PI and were negative for BrdUTP binding, while the upper right quadrant (R2) represents the nonviable (necrotic cells) which is positive for BrdUTP binding and showing PI uptake. The lower right quadrant (R3) represents the number of cells that had DNA fragmentation (apoptotic cells) and BrdUTP positive but negative for PI stain, indicating BrdUTP binding. The BrdUTP + PI apoptotic cell population for MDA-MB-231 cell line increased significantly (*P < 0.05) from 1.71 ± 0.02% in untreated cells to 11.66 ± 0.91% in cells treated for 24 hours and to 36.98 ± 2.71% and 44.52 ± 10.13% in cells treated for 48 and 72 hours (Table 1), respectively.

Bottom Line: Surgery, radiotherapy, and chemotherapy are the available treatments for breast cancer, but these were reported to have side effects.Morphological and apoptotic-inducing effects of NDV on MD-MB-231 cells were observed using phase contrast and fluorescence microscopes.It appears that NDV AF2240 strain is a potent anticancer agent that induced apoptosis in time-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratory of Anatomy & Histology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia ; Department of Human Anatomy, Faculty of Medicine, Bauchi State University, PMB 65, Gadau, Nigeria.

ABSTRACT
Breast cancer is the malignant tumour that developed from cells of the breast and is the first leading cause of cancer death among women worldwide. Surgery, radiotherapy, and chemotherapy are the available treatments for breast cancer, but these were reported to have side effects. Newcastle disease virus (NDV) known as Avian paramyxovirus type-1 (APMV1) belongs to the genus Avulavirus in a family Paramyxoviridae. NDV is shown to be a promising anticancer agent, killing tumour cells while sparing normal cells unharmed. In this study, the oncolytic and cytotoxic activities of NDV AF2240 strain were evaluated on MDA-MB-231, human mammary carcinoma cell line, using MTT assay, and its inhibitory effects were further studied using proliferation and migration assays. Morphological and apoptotic-inducing effects of NDV on MD-MB-231 cells were observed using phase contrast and fluorescence microscopes. Detection of DNA fragmentation was done following terminal deoxyribonucleotide transferase-mediated Br-dUTP nick end labeling staining (TUNEL) assay, which confirmed that the mode of death was through apoptosis and was quantified by flow cytometry. Furthermore, analysis of cellular DNA content demonstrated that the virus caused an increase in the sub-G1 phase (apoptotic peak) of the cell cycle. It appears that NDV AF2240 strain is a potent anticancer agent that induced apoptosis in time-dependent manner.

Show MeSH
Related in: MedlinePlus