Inhibitory effect of siRNA targeting CK2αgene on growth of HCT1 16 cells and its mechanism / 吉林大学学报(医学版)

ABSTRACT

Objective To investigate the inhibitory effect of siRNA targeting casein kinase 2 (CK2α)gene on the growth of HCT1 1 6 cells and to clarify its mechanism.Methods CK2α-siRNA sequence was designed according to mRNA sequence of CK2α. The in vitro cultured HCT1 1 6 cells were divided into normal control group (without transfection),negative control group (transfected with siRNA)and CK2α-siRNA group (transfected with CK2α-siRNA ),and the HCT116 cells were transfected with Lipofectamine 2000.The expression levels of CK2α,cyclin H,P53,and P21 proteins in the HCT116 cells were detected by Western blottting method,and the proliferation activities of the HCT116 cells were detected by MTT method,and the cell cycle was measured by flow cytometry. Results Compared with negative control group,the expression levels of CK2α,and cyclin H proteins in CK2α-siRNA group were decreased(P0.05), and the expression level of P21 protein was increased significantly(P<0.01).Compared with negative control group,the survival rate in CK2α-siRNA group was decreased markedly 48 and 72 h after transfection detected by MTT method(P<0.01).Flow cytometry analysis showed the percent of the cells at G1 phase in CK2α-siRNA group was significantly higher than that in negative control group and the percent of the cells at S phase in CK2α-siRNA group was lower than that in negative control group(P<0.01),and the cell cycle was arrested at G1 phase. Conclusion siRNA targeting CK2αcan inhibit the proliferation of HCT116 cells and induce the arrest of G1 phase, which may be associated with inhibiting the expression of cyclin H and recovering the P53 activity after silencing CK2α.