DEK基因在肝癌中的表达及靶向抑制其表达对肝癌细胞增殖及凋亡的影响

Expression of DEK gene in hepatocellular carcinoma and the effect of targeted inhibition gene expression on the proliferation and apoptosis of hepatocellular carcinoma cells

目的探讨DEK基因在肝癌中的表达及RNA干扰抑制其表达对肝癌细胞增殖及凋亡的影响及机制。方法以人正常肝细胞HL-7702作为对照,RT-PCR检测人肝癌HepG2、Huh-7、SMMC-7721、SNU-475细胞中DEK mRNA表达;DEK-siRNA转染HepG2细胞,同时转染阴性对照和空白对照,转染48 h后Western blotting检测各组细胞中DEK、Cleaved caspase 3、β-catenin、Cyclin D1蛋白表达;CCK8实验和流式细胞仪分别检测细胞的增殖及凋亡情况。结果各个肝癌细胞中DEK mRNA表达均显著高于正常细胞,差异有统计学意义(P<0.01);DEK基因在HepG2细胞中表达最高,选择作为后续研究对象;转染DEK-siRNA后DEK蛋白表达显著降低;DEK-siRNA组细胞存活率及β-catenin、Cyclin D1蛋白表达显著低于空白对照组和阴性对照组,细胞凋亡率和Cleaved caspase 3蛋白表达显著高于空白对照组和阴性对照组,差异有统计学意义(P<0.01)。结论抑制肝癌HepG2细胞中DEK基因可降低癌细胞的增殖及诱导细胞凋亡,其机制与Wnt/β-catenin信号通路的调控有关。

Objective To investigate the expression of RNA interference on the proliferation and apoptosis of HCC DEK in hepatocellular carcinoma （HCC） and the effect of cells. Methods Human normal liver cell line HL-7702 was as the control, expressions of DEK gene mRNA in human HCC HepG2, Huh-7, SMMC-7721, SNU-475 cells were detected by RT-PCR; DEK-siRNA was transfected into HepG2 cells, meanwhile negative control group and blank control group were transfected. Expressions of DEK, Cleaved caspase 3, β-catenin, Cyclin D1 protein after transfection for 48 hours were detected by Western blotting; CCK8 assay and flow cytometry were used to detect the proliferation and apoptosis of cells. Results The expression of DEK mRNA in various HCC cells was significantly higher than that in normal cells （P 〈0.01 ） , the expression of DEK gene was highest in HepG2 cells and it was selected as follow up study. The expression of DEK was significantly decreased after transfected DEK-siRNA. Cell survival rate and expressions of β- catenin, Cyclin D1 protein were significantly lower than those in blank control group and negative control group, the apoptosis rate and the expression of Cleaved caspase 3 protein were significantly higher than those in blank control group and negative control group （P 〈 0.01 ）. Conclusion DEK gene can inhibit the proliferation and induce apoptosis of hu- man HCC cell line HepG2, and its mechanism is related to the regulation of Wnt/β-catenin signaling pathway