抑癌因子miR-10a抑制Tiam1表达对胃癌细胞凋亡和迁移的影响

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。

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

ABSTRACT Objective: To investigate the effect of miR-10a 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 miR-10a expression and Tiam1 mRNA and protein expression levels in both cells. As well as, the expression of miR-10a and the expression of Tiam1 protein in gastric cancer cell S746T and normal gastric mucosal cells RGM-1 and NGEC were detected. By transfecting miR-10a mimic and miR-10a inhibitor into HS746T cells, flow cytometry was used to detect the cell cycle and apoptosis of HS746T, Transwell ^TM assay was used to detect the invasion ability of HS746T 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 miR-10a. The constructed Tiam1-highly expressed Tiam1-pcDNA3.1 plasmid and the PX458 plasmid knocked out of the Tiam1 gene were transfected into HS746T cells, respectively, and the apoptosis and invasion ability of HS746T cells were detected by flow cytometry and Transwell ^TM experiments. Results: Compared with gastric epithelial tissue cells, miR-10a expression was reduced in early gastric cancer clinical tissue cells, and expression of Tiam1 mRNA and protein was increased; The expression of miR-10a 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, miR-10a expression was reduced in gastric cancer cell HS746T, while Tiam1 protein expression was increased; miR-10a can inhibit the invasion of HS746T cells, promote apoptosis, and stagnate them in G0 / G1 phase; miR-10a targets the 3 ''untranslated region (3''UTR) of the Tiam1 gene and reduces Tiam1 protein expression; Tiam1 can inhibit the apoptosis of HS746T cells and promote the invasion of HS746T cells. Conclusion: miR-10a inhibits the proliferation and invasion of HS746T cells and promotes the apoptosis of HS746T cells by targeting the 3'' UTR of Tiam1 gene.