关键词:  , 酪蛋白激酶2, 结肠肿瘤, HCT116细胞, RNA干扰, 细胞周期

Abstract:

Objective To investigate the inhibitory effect of  siRNA targeting casein kinase 2 （CK2α） gene on the growth  of HCT116 cells and to clarify its mechanism.
Methods CK2α-siRNA sequence was designed according to mRNA sequence of CK2α.The in vitro cultured HCT116 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（P<0.01）;the expression level of P53 protein had no change dramatically（P>0.05）,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）.Fl
ow 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 CK2α can 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.

Key words: casein kinase 2, colon neoplasms, HCT116 cells, RNA interference, cell cycle