靶向着丝粒蛋白F的siRNA调节肝癌细胞增殖的实验研究

Experimental study on the regulation effect of siRNA targeting centromeric protein F on hepatoma cell proliferation

目的研究靶向着丝粒蛋白F(CENPF)的siRNA调节肝癌细胞增殖的作用及机制。方法培养肝癌细胞株中SMMC-7721、HepG2、BEL-7402及正常肝细胞株HL-7702,检测细胞中CENPF的表达水平;HepG2细胞进行分组处理,转染阴性对照(NC)siRNA或CENPF siRNA,给予对照溶剂二甲基亚砜(DMSO)或ERK1/2激动剂表皮生长因子(EGF),检测细胞增殖A490水平及细胞中CENPF、细胞周期蛋白D1(CyclinD1)、B细胞淋巴瘤/白血病-2基因(Bcl-2)、磷酸化细胞外调节蛋白激酶1/2(p-ERK1/2)的表达水平。结果肝癌细胞株中SMMC-7721、HepG2、BEL-7402中CENPF的表达水平高于正常肝细胞株HL-7702,差异有统计学意义(P<0.05);HepG2细胞中CENPF的表达水平最高;转染siRNA后,si-CENPF组HepG2细胞的A490水平及CENPF、CyclinD1、Bcl-2、p-ERK1/2的表达水平低于si-NC组,差异有统计学意义(P<0.05);转染siRNA的同时给予ERK1/2激动剂EGF,EGF+si-CENPF组HepG2细胞的A490水平及CENPF、CyclinD1、Bcl-2、p-ERK1/2的表达水平均高于DMSO+si-CENPF组,差异有统计学意义(P<0.05)。结论靶向CENPF的siRNA可显著抑制肝癌细胞的增殖,相关的分子机制可能是抑制ERK1/2的磷酸化并抑制下游CyclinD1、Bcl-2的表达。

Objective To study the effect and mechanism of siRNA targeting centromeric protein F(CENPF)on the proliferation of hepatoma cells.Methods Hepatoma cell line SMMC-7721,HepG2,BEL-7402 and normal hepatocyte line HL-7702 were cultured,and the expression of CENPF was detected.HepG2 cells were treated in groups,transfected with negative control(NC)siRNA or CENPF siRNA,and given control solvent dimethyl sulfoxide(DMSO)or ERK1/2 agonist epidermal growth factor(EGF),the level of cell proliferation A490 and the expression of CEN-PF,Cyclin D1,B-cell lymphoma/leukaemia-2 gene(Bcl-2),phosphorylated extracellular regulated protein kinase 1/2(p-ERK1/2)were detected.Results The expression level of CENPF in SMMC-7721,HepG2 and BEL-7402 was higher than that in HL-7702,and the difference was statistical-ly significant(P<0.05).The expression level of CENPF in HepG2 cells was the highest;after transfection with siRNA,the A490 level and the ex-pression levels of CENPF,CyclinD1,Bcl-2 and p-ERK1/2 of HepG2 cells in the si-CENPF group were lower than those in the si-NC group,and the differences were statistically significant(P<0.05);the A490 level and the expression levels of CENPF,CyclinD1,Bcl-2 and p-ERK1/2 of HepG2 cells in the EGF+si-CENPF group were higher than those in the DMSO+si-CENPF group,and the differences were statistically significant(P<0.05).Conclusion siRNA targeting CENPF significantly inhibit the proliferation of hepatoma cells,the related molecular mechanism may be the in-hibition of the phosphorylation of ERK1/2 and the expression of downstream CyclinD1 and Bcl-2.