In vitro and in vivo evaluation of the cyclophosphamide effect along with oncolytic Newcastle disease virus (NDV): An animal preclinical research

Abstract

Background: Breast cancer is a major cause of mortality globally. Oncolytic virotherapy is a promising treatment modality that directly destroys cancer cells and induces an immune response against them. Among natural oncolytic viruses, Newcastle disease virus (NDV) has shown selective tumor cell infection.

Materials and methods: In this study, we investigated the efficacy of variable doses of NDV and cyclophosphamide on 4T1 cancer cell line and BALB/c mouse tumors for the first time.

Results: Compared with the control group, the combination treatment group with NDV and cyclophosphamide showed a significant increase in the expression levels of P21, P27, and P53 genes by 38%, 46%, and 81%, respectively (p < 0.05). In contrast, the expression levels of CD34, integrin α5, vascular endothelial growth factor (VEGF), and vascular endothelial growth factor receptor (VEGFR) genes significantly decreased by 47%, 45%, 42%, and 23%, respectively (p < 0.05). The reactive oxygen species (ROS) generation assay evaluated with 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining showed a significant increase in ROS levels within 4T1 cells treated with NDV compared with the untreated group after 24 h (p < 0.01). Furthermore, Annexin V/propidium iodide (PI) double staining analysis showed that the proportion of apoptotic cells in the NDV-treated group decreased by 0.61% and 1.63% after 6 h and 12 h, respectively (p < 0.05). After 12 days, tumor volume in the NDV-treated groups decreased by 72%−87% compared with a 48% increase in the control group, reflecting a net reduction in tumor volume relative to the control group (p < 0.001).

Conclusion: These findings demonstrate that NDV in combination with chemotherapy drugs may be a promising therapeutic option for cancer patients. However, several other factors need to be considered. These results indicate that NDV may have potential effects on cancer treatment.

Keywords

breast cancer / Newcastle disease virus / apoptosis / noninvasive treatment / 4T1 cell line

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