Effect of enhanced survivin expression on lymphocyte proliferation and function in K562 cells / 第三军医大学学报

ABSTRACT

Objective To study the effect of survivin gene expression on the lymphocyte’s proliferation and function in cultured K562 cells. Methods The constructed recombinant vector pEGFP-C1-survivin and the plasmid pTZU6+1-survivin encoding short hairpin RNA of survivin were transfected into K562 cells respectively to generate K562/survivin+ cells and K562/survivin-cells. K562/survivin+ cells were selected by G418. The survivin mRNA and protein levels in the 3 kinds of cells (K562/survivin+,K562 and K562/survivin-) were detected by semi-quantitative RT-PCR and immunohistochemistry methods. The peripheral blood mononuclear cells (PBMC) from healthy subjects were co-cultured with these 3 cells respectively in mixed lymphocyte-tumor cell culture (MLTC). Lymphocyte proliferation in the supernatant was evaluated by MTT assay. Nature killer activity was detected by flow cytometry and IFN-? level was measured by ELISA assay. Results Proliferation index in K562/survivin+ group was much lower than that in the other 2 groups (P