Abstract: Objective To construct c-Met target chimeric antigen receptors(chimeric antigen receptor, CAR)retrovirus vector and to explore its expression and effects of modified T lymphocytes on the c-Met positive nasopharyngeal carcinoma cells. Methods Genetic engineering method was used to construct the c-Met scFv, and then the c-Met scFv was inserted into retrovirus vector containing signaling molecules such as CD28, CD137 and CD3ζ.The build c-Met CAR retrovirus vector was confirmed by sequencing. Western blotting was used to analyze c-Met CAR expression in T lymphocyte after the virus vector infected T lymphocyte. CCK8 assay was employed to detect the cytotoxicity against the nasopharyngeal carcinoma cell lines, and changes of IFN-γ and IL-2 was analyzed by ELISA after c-Met CAR-T co-culture with nasopharyngeal carcinoma cells. Results The sequencing result showed that the c-Met CAR vector sequences were right. Western blotting could detect the expression of CD3ζ. CCK-8 assay indicated that c-Met CAR-T cells suppressed proliferation of nasopharyngeal carcinoma cells, and ELISA assay demonstrated that c-Met CAR-T cells could effectively release IFN-γ and IL-2 with the help of nasopharyngeal carcinoma cells. Conclusion c-Met CAR retrovirus vector was established successfully. The CAR could be expressed by the vector in T cells. c-Met CAR-T inhibited proliferation of c-Met positive nasopharyngeal carcinoma cells and increased the secretion of IFN-γ and IL-2.

Key words: Nasopharyngeal carcinoma, c-Met, Chimeric antigen receptor