eIF4A1基因沉默对胃癌SGC-7901细胞生物学行为影响机制的探讨  

Effect of eIF4A1 gene silencing on biological behavior of SGC-7901 in gastric cancer

作　　者：谢东毅 韦尉元 吴锟[1] 谢玉波[3] 肖强[1] XIE Dong-yi;WEI Wei-yuan;WU Kun;XIE Yubo;XIAO Qiang(Department of Gastrointestinal and Gland Surgery,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,南宁530021 [2]广西医科大学附属肿瘤医院胃肠外科,南宁530021 [3]广西医科大学第一附属医院麻醉科,南宁530021

出　　处：《中国临床新医学》2018年第4期315-320,共6页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：国家自然科学基金资助项目(编号:81660511);广西自然科学基金重点项目(编号:2015GXNSFDA227001)

摘　　要：目的探讨沉默eIF4A1基因对胃癌细胞增殖、凋亡、迁移的细胞生物学行为影响的机制。方法以eIF4A1沉默重组慢病毒颗粒(LV-EIF4A1-RNAi组)感染人胃癌SGC-7901为eIF4A1沉默组(eIF4A1-RNAi组),感染阴性对照慢病毒颗粒(CON077)的胃癌细胞为阴性对照组(CON077组),空白对照组(control组)不作任何处理。以每个副孔初始含4×10~3个细胞,CCK-8检测各组细胞增殖能力;流式细胞仪检测各组5×10~6个细胞凋亡和1×10~6个细胞周期分布;Transwell实验检测各组1×10~5个细胞迁移和侵袭能力;实时荧光定量PCR和Western blot检测各组细胞eIF4A1和AKT基因的mRNA和蛋白表达水平。结果 LV-EIF4A1-RNAi(44683-1)组eIF4A1基因的表达量最低为(0.31±0.04),因此选择该组为eIF4A1沉默组(eIF4A1-RNAi)。与CON077组和control组相比,eIF4A1-RNAi组细胞增殖活力明显降低,细胞凋亡率提高,细胞周期阻滞于S期,G1/G2期细胞减少,细胞的迁移、侵袭能力下降,eIF4A1和AKT基因的mRNA和蛋白表达明显降低,差异均有统计学意义(P<0.05);CON077组与control组比较,上述指标差异均无统计学意义(P>0.05)。结论慢病毒介导eIF4A1基因沉默能抑制人胃癌细胞株SGC-7901的增殖和迁移、促进细胞凋亡、阻滞细胞周期于S期,其机制可能与AKT表达下调有关。Objective To investigate the effects of eIF4A1 gene silencing on the proliferation,apoptosis,migration of human gastric cancer cell lines and its molecular mechanisms.Methods Human gastric cancer cell lines SGC-7901 were infected with the recombinant lentivirus vector LV-EIF4A1-RNAi for eIF4A1 gene silencing,as eIF4A1-RNAi group,while control recombinant lentivirus vector CON077 infected with human gastric cancer cell lines SGC-7901,as the negative control group(CON077)and blank control group without any treatment.CCK-8 was used to detect cell proliferation activity,and the cell apoptosis rate and cell cycle distribution were detected by flow cytometry.The activity of migration and invasion of cells were evaluated by Transwell.Quantitative real-time polymerase chain reaction(QRT-PCR)and Western blot were applied for measurement of mRNA and protein expressions of eIF4A1 and AKT.Results The expression of eIF4A1 gene in LV-EIF4A1-RNAi(44683-1)group was the lowest(0.31±0.04),so the group was selected as eIF4A1 silent group(eIF4A1-RNAi).Compared with those in the CON077 group and the control group,the cell proliferation activity was significantly decreased,the cell apoptosis rate was increased,the cell cycle was blocked in S phase,the G 1/G 2 phase cells were decreased,and the cell migration and invasion ability were decreased in the eIF4A1-RNAi group(P<0.05).The expressions of mRNA and protein of eIF4A1 and AKT were down-regulated.No significant differences were found in the measurements between the CON077 group and the blank control group(P>0.05).Conclusion Lentivirus-mediated eIF4A1 gene silencing inhibits the growth and migration of human gastric cancer cell lines,induces cells apoptosis and arrests cell cycle in S phase,and its mechanisms may be related to the downregulation of AKT expression.

关 键 词：胃癌 eIF4A1 细胞增殖 细胞凋亡 细胞迁移 AKT 

分 类 号：R735.2[医药卫生—肿瘤]