微小RNA-199对胃癌细胞增殖和迁移的影响  

Effect of MicroRNA-199 on Proliferation and Migration of Gastric Carcinoma Cells

作　　者：闫超[1] 张连海[1] 陕飞[1] 李双喜[1] 贾永宁[1] 李子禹[1] YAN Chao;ZHANG Lianhai;SHAN Fei;LI Shuangxi;JIA Yongning;LI Ziyu(Key Laboratory of Carcinogenesis and Translational Research(Ministry of Education/Beijing),Gastrointestinal Cancer Center, Peking University Cancer Hospital and Institute,Beijing 100142,China)

机构地区：[1]北京大学肿瘤医院暨北京市肿瘤防治研究所,胃肠肿瘤中心,恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142

出　　处：《中国医学科学院学报》2018年第4期528-533,共6页Acta Academiae Medicinae Sinicae

基　　金：科技部国家科技支撑计划(2014BAI09B02)、北京市医院管理局临床医学发展专项(ZYLX201701)和北京市医院管理局“使命”计划专项(SML20151001).

摘　　要：目的检测微小RNA(miR)-199在胃癌组织以及胃癌细胞株的表达,探讨miR-199对胃癌细胞增殖和迁移的影响。方法采用逆转录-聚合酶链反应技术检测miR-199在51例胃癌组织和配对癌旁组织以及胃癌细胞株和正常胃黏膜上皮细胞GES-1中的表达,建立miR-199过表达或低表达的胃癌细胞株,采用细胞增殖实验、细胞迁移实验、蛋白免疫印迹技术检测胃癌细胞的增殖、迁移能力的变化及其相关机制。结果 miR-199在胃癌组织中的相对表达量显著低于配对的癌旁组织,差异有统计学意义(0.2635±0.0303比1.6700±0.9613,t=13.95,P<0.001);在胃癌细胞株AGS(0.81,t=9.13,P<0.001)、SGC-7901(0.83,t=8.88,P<0.001)、MKN28(0.58,t=10.80,P<0.001)、KATO-Ⅲ(0.60,t=10.31,P<0.001)、MKN-45(0.27,t=13.10,P<0.001)中的相对表达量显著低于正常胃黏膜上皮细胞GES-1(2.1),差异有统计学意义;在胃癌细胞株MKN-45中,高表达miR-199组细胞的增殖能力、迁移能力较对照组胃癌细胞显著降低,差异有统计学意义(731±13比345±18,t=24.90,P<0.001);在胃癌细胞KATO-Ⅲ中,低表达miR-199组细胞的增殖能力、迁移能力较对照组胃癌细胞显著增强,差异有统计学意义(257±16比657±8,t=32.59,P<0.001)。双荧光素酶报告实验显示miR-199可以直接作用于TBL1XR1的3’非翻译区,调控TBL1XR1的表达。Western blot结果显示高表达miR-199后,TBL1XR1表达水平降低。结论 miR-199在胃癌组织中呈低表达,可能与胃癌细胞的增殖和迁移能力有关,miR-199的作用可能是通过调控TBL1XR1实现的。Objective To detect the expression of microRNA(miR)-199 in gastric carcinoma tissues and cell lines,and further explore the effect and molecular mechanism of miR-199 on the proliferation and migration of gastric carcinoma cell lines.Methods Reverse transcriptase-polymerse chain reaction was used to detect the expression of miR-199 in gastric carcinoma and adjacent normal tissue obtained from 51 patients and in gastric carcinoma cell lines and human gastric epithelial cell line GES-1.The gastric carcinoma cell lines over-expressing and low-expressing miR-199 were established to detect their proliferation and migration abilities.Dual-luciferase reporter assay was performed to detect the regulatory effect of miR-199 on the 3’untranslated region of TBL1XR1.Western blot was used to explore the miR-199-related mechanism.Results The relative expression of miR-199 in gastric carcinoma tissues was significantly lower than that in the adjacent normal tissue(0.2635±0.0303 vs.1.6700±0.9613,t=13.95,P<0.001).The relative expressions of miR-199 in gastric carcinoma cell lines AGS(0.81,t=9.13,P<0.001),SGC-7901(0.83,t=8.88,P<0.001),MKN28(0.58,t=10.80,P<0.001),KATO-Ⅲ(0.60,t=10.31,P<0.001),MKN-45(0.27,t=13.10,P<0.001)were significantly lower than that in the normal gastric cell line GES-1(2.1).In miR-199 over-expressed cell lines,the cell proliferation and migration significantly decreased as compared with the control group of gastric carcinoma cells(731±13 vs.345±18,t=24.90,P<0.001),and in miR-199 low-expressed group,the cell proliferation and migration increased compared with the control group of gastric carcinoma cells(257±16 vs.657±8,t=32.59,P<0.001).Dual-luciferase reporter assay proved that miR-199 directly targeted on the 3’untranslated region of TBL1XR1.Western blot analysis showed that miR-199 inhibited the expression of TBL1XR1.Conclusion The over-expression of miR-199 in gastric carcinoma is associated with the decreased ability of proliferation and migration of gastric carcinoma cells by targeting TBL1XR1.

关 键 词：胃癌 微小RNA-199 细胞增殖 细胞迁移 TBL1XR1 

分 类 号：R735.2[医药卫生—肿瘤]