NDUFA4过表达对人结直肠癌细胞体外生长的影响

目的探讨过表达NDUFA4对人结直肠癌细胞体外生长的影响及其机制。方法将p-NDUFA4重组质粒(p-NDUFA4组)和p-Cont对照质粒(p-Cont组)体外分别转染人结直肠癌HCT116细胞;采用Real-timePCR和Westernblot检测细胞中NDUFA的表达;CCK-8法和克隆形成实验分别检测HCT116细胞增殖活性和克隆形成能力;实时荧光定量PCR法检测HCT116细胞中CDK2、CDK3、CDK4、CDK6的mRNA水平;Westernblot法检测细胞中蛋白总AKT、ERK1/2以及磷酸化AKT(p-AKT)、ERK1/2(p-ERK1/2)的表达水平。结果与p-Cont对照组相比,p-NDUFA4组中NDUFA4的表达水平显著升高(均P<0.01);细胞增殖和克隆形成能力明显增强(均P<0.05);CDK2、CDK3等的mRNA水平显著上调(均P<0.05);p-NDUFA4组中p-AKT和p-ERK1/2蛋白的表达水平明显上调(均P<0.05)。结论过表达NDUFA4能明显促进人结直肠癌细胞的体外生长,其机制可能与AKT和ERK信号通路的变化相关。

Effects of NDUFA4 Overexpression on Growth of Human Colon Cancer Cells in vitro

Objective To investigate the effects of NDUFA4 overexpression on the growth of human colon cancer cells in vitro and to explore its potential mechanism. Methods The recombinant plasmid p-NDUFA4 (p-NDUFA4) and the control plasmid (p-Cont) were transiently transfected into human colon cancer HCT 116 cells, respectively. Then, the relative expression level of NDUFA4 was determined by Real-time PCR and Western blot. The proliferation and colony formation ability of HCT116 cells were detected by CCK-8 assay and clone formation assay. The mRNA expression levels of cyclin-dependent kinase (CDK) 2, CDK3, CDK4 and CDK6 were determined by Real-time PCR. The expression levels of AKT (p-AKT) and ERK1/2(p-ERK1/2) were analyzed by Western blot. Results Compared with control group, the expression levels of NDUFA4 were increased remarkably in p-NDUFA4 group (all P<0.01). The proliferation and clone formation capacity of HCT116 cells in p-NDUFA4 group were obviously increased (all P<0.05). The expression of CDK2 and CDK3 mRNA were increased significantly(all P<0.05). The expression of p-AKT and p-ERK1/2 were increased obviously (all P<0.05). Conclusion The overexpression of NDUFA4 could significantly promote the growth of human colon cancer HCT116 cells in vitro, which might be closely related to the change of AKT and ERK signaling transduction.