GPD2基因敲减对肺癌细胞H1299生物学功能的影响

Effect of GPD2 gene knockdown by shRNA on the biological function of lung cancer cells H1299

目的:沉默甘油-3-磷酸脱氢酶2(GPD2)基因表达,验证对肺癌细胞增殖、凋亡、侵袭等生物功能及行为学的影响。方法:构建GPD2基因的RNA干扰慢病毒载体(shGPD2)和对应的对照载体(shCtrl),并感染H1299细胞,实时荧光定量(PCR)检测GPD2基因在癌细胞中的mRNA变化情况,Western blotting检测GPD2基因在癌细胞中蛋白质变化情况,验证敲减效率;CCK8法检测细胞的增殖情况,流式细胞术检测细胞凋亡情况;Transwell实验检测细胞侵袭情况。结果:成功构建慢病毒干扰载体shGPD2并感染H1299细胞,mRNA及蛋白质含量均减少,其敲减效率达到73.5%(P<0.05);GPD2基因敲减后的H1299细胞增殖明显减缓,细胞凋亡显著增多,细胞侵袭能力明显下降(P<0.01)。结论:GPD2基因敲减能够有效抑制肺癌细胞H1299的增殖。

Objective:To investigate the effects of silencing the expression of glycerol-3-phosphate dehydrogenase 2(GPD2)gene on the biological function(proliferation,apoptosis and invasion)and behavior of lung cancer cells.Methods:The RNA interference lentiviral vector containing GPD2 gene(shGPD2)and corresponding control vector(shCtrl)were constructed,and used to infect the H1299 cells.The expression levels of GPD2 gene mRNA and protein in cancer cells were detected using qPCR and Western blotting,respectively.The knockdown efficiency of GPD2 gene was verified.The proliferation,apoptosis and invasion of cells were detected using CCK8 method,flow cytometry and Transwell assay,respectively.Results:The lentiviral vector shGPD2 was successfully constructed,and infected H1299 cells.The mRNA and protein levels decreased,and the knockdown efficiency was 73.5%(P<0.05).After the GPD2 gene expression decreased,the proliferation,apoptosis and invasion ability of H1299 cells significantly slowed down,increased and reduced,respectively(P<0.01).Conclusions:The GPD2 gene knockdown can effectively inhibit the proliferation of lung cancer cells H1299.