过表达miR-30b对人胃癌细胞株SGC-7901和AGS生物学功能及其肿瘤形成的影响  

Effects of overexpression of miR-30b on the biological function and tumor formation of human gastric cancer cells

作　　者：陈翠翠 梁焕坤 李康燕 程承武 刘细潘 黎杰星 钟树海 张立成[2] 李来庆 CHEN Cui-cui;LIANG Huan-kun;LI Kang-yan;CHENG Cheng-wu;LIU Xi-pan;LI Jie-xing;ZHONG Shu-hai;ZHANG Li-cheng;LI Lai-qing(Guangzhou Youdi Biotechnology Co., Ltd., Guangzhou 510663, China;Department of Oncology, Eighty-eighth Hospital of PLA)

机构地区：[1]广州优迪生物科技股份有限公司,510663 [2]解放军第88医院肿瘤科

出　　处：《天津医药》2017年第7期677-681,共5页Tianjin Medical Journal

基　　金：广东省科技计划项目(2016A010119067);山东省医药卫生科技发展计划项目(2016WS0725)

摘　　要：目的研究过表达miR-30b对人胃癌细胞株SGC-7901和AGS的增殖、细胞周期、凋亡、侵袭等生物学功能的影响及对体内肿瘤形成的抑制作用。方法 SGC-7901和AGS细胞中转染miR-30b类似物和miR-对照后,qRT-PCR检测转染后细胞miR-30b的表达变化;Western blot检测eIF5A2蛋白的表达;CCK-8检测SGC-7901和AGS细胞增殖;流式细胞术检测细胞周期和凋亡;Transwell实验检测细胞体外侵袭能力。转染miR-30b类似物和miR-对照的SGC-7901和AGS细胞,分别注射裸鼠体内,观察肿瘤的形成及肿瘤组织中eIF5A2蛋白的表达。结果qRT-PCR结果表明miR-30b类似物组的相对表达量显著高于miR-对照组(P<0.05);Western blot结果显示,miR-30b类似物组的eIF5A2蛋白表达降低;CCK-8实验表明miR-30b类似物组中细胞增殖受到抑制;流式细胞术显示miR-30b类似物组的细胞周期减慢,凋亡增加;在Transwell迁移实验中,miR-30b类似物组细胞侵袭能力明显低于miR-对照组(P<0.05)。细胞体内成瘤实验显示,miR-30b过表达抑制肿瘤的形成,肿瘤组织中eIF5A2蛋白表达降低。结论 miR-30b过表达抑制人胃癌细胞的增殖、侵袭和肿瘤的形成,降低eIF5A2蛋白的表达,为胃癌治疗提供了一个潜在的靶点。Objective To investigate the effect of overexpression of miR-30b on the proliferation,cell cycle,apoptosis and invasion of gastric cancer cell line SGC-7901and AGS,and the inhibitory effect on the tumor formation in vivo.Methods SGC-7901and AGS cells were transfected with miR-30b mimics and miR-control,and qRT-PCR was used to detect the expression levels of miR-30b.Western blot assay was used to detect the expression of eIF5A2protein.CCK-8assay was used to measure the cell proliferation.Flow cytometry was used to analyze cell cycle and apoptosis.Transwell assay was used to detect cell invasion.In addition,the SGC-7901and AGS cells transfected with miR-30b mimics and miRcontrol were injected into nude mice to observe the tumor formation and the expression of eIF5A2protein in vivo.Results Results of qRT-PCR showed that the relative expression of miR-30b was significantly higher than that of miR-control group(P<0.05).Western blot assay showed that the expression of eIF5A2protein was decreased in miR-30b mimics group.CCK-8assay showed that cell proliferation was inhibited in miR-30b mimics group.The result of flow cytometry showed that the cell cycle decreased and the apoptosis increased in miR-30b group.Transwell assay showed that the cell invasion was significantly lower in miR-30b group than that of control group(P<0.05).Overexpression of miR-30b inhibited the formation of tumor and decreased the expression of eIF5A2protein in vivo.Conclusion Overexpression of miR-30b inhibits the proliferation,invasion and tumor formation of gastric cancer cells,and reduces the expression of eIF5A2protein,which provides a potential target for gastric cancer treatment.

关 键 词：微RNAS 细胞增殖 细胞周期 细胞凋亡 肿瘤侵润 miR-30b 

分 类 号：R573.9[医药卫生—消化系统]