抑癌因子miR-10a抑制Tiam1表达对胃癌细胞凋亡和迁移的影响  

Inhibition of Tiam1 Expression by Tumor Suppressor miR-10a on Proliferation and Migration of Gastric Cancer Cells

作　　者：吴江[1] 阿里木江•阿不都热合曼 庞澜[1] 朱勇荷 马秀英[1] 陈鹏[1] 张荔霜[1] 岳跃明 WU Jiang;ALIMUJIANG·Abudoureheman;PANG Lan;ZHU Yong-he;MA Xiu-ying;CHEN Peng;ZHANG Li-shuang;YUE Yue-ming(The Second Affiliated Hospital of Xinjiang Medical University,Endoscopic diagnosis and treatment,Urumqi,Xinjiang,830063,China)

机构地区：[1]新疆医科大学第二附属医院内镜诊治一科,新疆乌鲁木齐830063

出　　处：《现代生物医学进展》2020年第15期2830-2837,2867,共9页Progress in Modern Biomedicine

基　　金：新疆维吾尔自治区自然科学基金项目(2016D01C201)。

摘　　要：目的:探讨miR-10a抑制Tiam1表达对胃癌细胞凋亡和侵袭的影响。方法:获取胃上皮组织细胞及胃癌组织细胞,利用q PCR及Western blot实验检测两种细胞中mi R-10a表达与Tiam1的m RNA及蛋白表达水平,同时检测胃癌细胞S746T及正常胃粘膜细胞RGM-1和NGEC中mi R-10a表达与Tiam1蛋白表达水平。通过将mi R-10a mimic和mi R-10a inhibitor转染HS746T细胞,利用流式细胞术检测HS746T的细胞周期和细胞凋亡,TranswellTM实验检测HS746T细胞的侵袭能力,qPCR及Western blot实验检测凋亡相关蛋白caspase3、caspase9和Bax以及周期相关蛋白P21表达水平;荧光素酶活性分析实验检测Tiam1是mi R-10a的作用靶点。已构建的Tiam1高表达的Tiam1-pcDNA3.1质粒和敲除Tiam1基因的PX458质粒分别转染HS746T细胞,通过流式细胞术及TranswellTM实验检测HS746T细胞的凋亡及侵袭能力。结果:与胃上皮组织细胞相比,早期胃癌临床组织细胞中mi R-10a表达降低,Tiam1的m RNA及蛋白表达升高;mi R-10a的表达与早期胃癌患者的肿瘤转移密切相关,与年龄、性别和肿瘤分期无关;与正常胃粘膜细胞RGM-1和NGEC相比,胃癌细胞HS746T中的mi R-10a表达降低,而Tiam1蛋白表达升高;mi R-10a可抑制HS746T细胞侵袭,促进细胞凋亡,使其停滞于G0/G1期;mi R-10a靶向作用于Tiam1基因的3’非翻译区(3’UTR),减少Tiam1的蛋白表达;Tiam1可抑制HS746T细胞凋亡,促进HS746T细胞侵袭。结论:mi R-10a靶向作用于Tiam1基因的3’UTR,抑制HS746T细胞的增殖及侵袭,促进HS746T细胞凋亡。Objective: To investigate the effect of mi R-10 a inhibition of Tiam1 expression on apoptosis and invasion of gastric cancer cells. Methods: Obtain gastric epithelial tissue cells and gastric cancer tissue cells. QPCR and Western blot experiments were used to detect mi R-10 a expression and Tiam1 m RNA and protein expression levels in both cells. As well as, the expression of mi R-10 a and the expression of Tiam1 protein in gastric cancer cell S746 T and normal gastric mucosal cells RGM-1 and NGEC were detected. By transfecting mi R-10 a mimic and mi R-10 a inhibitor into HS746 T cells, flow cytometry was used to detect the cell cycle and apoptosis of HS746 T, TranswellTMassay was used to detect the invasion ability of HS746 T cells, and qPCR and Western blot were used to detect the apoptosis-related protein caspase3, Caspase9, Bax, and cycle-associated protein P21 expression levels;luciferase activity analysis experiments detected Tiam1 as the target of mi R-10 a. The constructed Tiam1-highly expressed Tiam1-pcDNA3.1 plasmid and the PX458 plasmid knocked out of the Tiam1 gene were transfected into HS746 T cells, respectively, and the apoptosis and invasion ability of HS746 T cells were detected by flow cytometry and TranswellTMexperiments. Results: Compared with gastric epithelial tissue cells,mi R-10 a expression was reduced in early gastric cancer clinical tissue cells, and expression of Tiam1 m RNA and protein was increased;The expression of mi R-10 a is closely related to tumor metastasis in patients with early gastric cancer, and has nothing to do with age,gender and tumor stage;Compared with normal gastric mucosal cells RGM-1 and NGEC, mi R-10 a expression was reduced in gastric cancer cell HS746 T, while Tiam1 protein expression was increased;mi R-10 a can inhibit the invasion of HS746 T cells, promote apoptosis,and stagnate them in G0/G1 phase;mi R-10 a targets the 3 ’untranslated region(3’UTR) of the Tiam1 gene and reduces Tiam1 protein expression;Tiam1 can inhibit the apoptosis of HS746 T cells and

关 键 词：胃癌 miR-10a TIAM1 细胞凋亡 细胞侵袭 

分 类 号：R-33[医药卫生] R735.2