Production of carcinoma-targeting newcastle disease virus by propagation in bhk cells expressing hea125 membrane protein

Oncolytic viruses are natural or genetically modified viral species that preferentially infect and kill cancer cells. One of such viruses is the Newcastle Disease Virus (NDV). NDV has a natural preferencefor replication in a variety of tumor cells as opposed to normal tissue. Carcinomas (epithelial cancers) are the most common type of cancer in Iran and the world. Epithelial cell adhesion molecule (EpCAMCD326) is one of the dominant tumor antigens of epithelial origin. EpCAM has high expression and notable distribution on the membrane of cancer cell, and may, therefore, serve as a diagnostic marker for related cancers and also as a convenient receptor for drugs delivery. Thus, production of NDV with anti-EPCAM antibody in its envelope can contribute to treatment of all carcinomas. BHK-HEA125 cells (Baby Hamester Kidney which containing anti-EpCAM antibody - HEA125- in the membrane) were infected with NDV. After 4 days, the NDVs, which had replicated in the cells and acquired anti-EpCAM antibody (HEA125) by budding from the cell membrane, were collected and verified by HA and HI tests. The production of sarcoma-targeted virus was measured by ELISA method and the effect of virus proliferation on carcinoma (SK-BR-3 cell line) was evaluated by cell viability assay using flow cytometry and XTT-assay. After propagation of NDV V4-strain in the cells which stable transfected with plasmid encoding EpCAM antibody, titration was performed by HA using the standard method and the minimum titre of the virus was 160HU. The OD reading in the ELISA test showed significant sign of production of mosaic virus with HEA125 on the envelope. The results of flow cytometry and XTT-assay indicated that the mosaic virus had a greater effect on carcinoma cells than did the wild strain. The wild strain of NDV has good proliferation and CPE in tumor cells; but some results indicate that the mosaic virus with HEA125 has a better effect than the wild strain. In the future, this virus can be used in in-vivo studies, and in the case of desirable results and after appropriate pre-clinical studies, this approach can be used in clinical studies to make NDV target all carcinomas as well as metastases derived from colon, stomach, pancreas, liver, lung, breast, ovary, thyroid gland, kidney, bladder and prostate cancers

کلیدواژه ها:

Breast Cancer ، Gene and Cancer ، Cell and Cancer ، Targeted Cancer Therapy ، Drugs and Cancer