Overexpression of human CKLF2 promotes proliferation of murine BALB/c 3T3 fibroblast cells and inhibits their apoptosis

Objective To study the effect of human chemokine-like factor 2(CKLF2)on the proliferation and cell survival of BALB/c 3T3 cells. Methods We constructed a eukaryotic expression vector of pcDI/CKLF2 and established two cell lines, including CKLF2-overexpressed cell line (3T3/CKLF2) and a control cell line(3T3/pcDI). Cell proliferation was analyzed by MTT and cell counting assay; cell apoptosis after serum-withdrawal was analyzed by annexin-Ⅴ staining. Results We have successfully established two cell lines, 3T3/CKLF2 and 3T3/pcDI, and identified expression of CKLF2 by RT-PCR; 3T3/CKLF2 cells grow faster than the control cells, after cell culture for 40h, the number of 3T3/CKLF2 is 2-3 times more than that of 3T3/pcDI and wild type cells. Flow cytometry analysis showed that annexin-Ⅴ positive cells in 3T3/CKLF2 were less than those in 3T3/pcDI cells after serum withdrawal. Conclusion CKLF2 could promote proliferation and exert an anti-apoptotic effect on BALB/c 3T3 cells. These results provide experimental evidences for further study of biological activity of CKLF family. [