张雪莲,吴维宇,何峰
ZHANG Xuelian,WU Weiyu,HU Feng

• 1:成都医学院第一附属医院消化内科

摘要(Abstract)：

目的:探索miR-10b在胃癌细胞中的表达及对胃癌细胞增殖和凋亡的影响及相关机制。方法:选取20例确诊胃癌患者,取胃癌组织标本及其癌旁正常组织标本,采用实时荧光定量PCR(qRT-PCR)检测miR-10b和上皮型钙黏蛋白(E-cadherin);利用Lip2000转染试剂盒说明书,将miR-10b转染至对数生长期的人胃癌细胞系,miR-NC作为空白对照组,qRT-PCR检测胃癌组织和细胞中miR-10b以及E-cadherin表达的变化; Western blot检测胃癌组织和细胞中E-cadherin和凋亡蛋白B淋巴细胞瘤-2基因(Bcl2)、Bcl2相关X蛋白(Bax)表达变化;噻唑蓝(MTT)法检测MKN-45细胞增殖能力的改变。结果:qRT-PCR结果显示,与癌旁组织比较,胃癌组织中miR-10b的表达升高,而E-cadherin的表达降低(P <0. 05);与空白对照组比较,低分化胃癌组织和MKN-45癌细胞miR-10b表达增高,E-cadherin表达降低,差异有统计学意义(P <0. 05);与空白对照组相比,转染miR-10b siRNA后,E-cadherin和Bax表达增加,Bcl2表达和细胞生长增殖抑制率降低,差异均有统计学意义(P <0. 05);与空白对照组相比,GES-1细胞转染miR-10b mimics后,细胞内E-cadherin和Bax表达下降,Bcl2表达和细胞生长增殖抑制率增加,差异均有统计学意义(P <0. 05)。结论:miR-10b能促进胃癌细胞增殖并抑制癌细胞的凋亡,其机制可能与调控E-cadherin有关。
Objective: To explore the expression of miR-10 b in gastric cancer cells and its effect on the proliferation and apoptosis of gastric cancer cells and its possible mechanism. Methods: 20 patients with gastric cancer were enrolled in this study. Gastric cancer tissue samples and matched normal tissue samples were collected. MiR-10 b and E-cadherin were detected by quantitative real-time PCR( qRT-PCR). The miR-NC( negative control) and miR-10 b were transfected into human gastric cancer cell line with logarithmic phase using the method provided by Invitrogen Lip2000 Transfection Kit. The qRT-PCR was used to detect the expression changes of gastric cancer cellular tissues and miR-10 b and E-cadherin. Western Blot was used to detect the expression changes of E-cadherin and apoptosis proteins Bcl2 and Bax in gastric cancer tissues and cells. MTT assay was used to detect the proliferation of MKN-45 cells. Results: The results of qRT-PCR showed that the expression of miR-10 b in gastric cancer tissues was significantly higher than that of the adjacent tissues,while the expression of E-cadherin significantly decreased( P < 0. 05). Compared with the control group,the expression of miR-10 b increased and the expression of E-cadherin decreased in poorly differentiated gastric cancer cells( P < 0. 05). The expression of E-cadherin increased after transfection of miR-10 b siRNA,and the expression of Bcl2 markedly decreased; the expression of Bax increased,and the cell proliferation inhibition rates significantly decreased as compared with the control group( P < 0. 05).After transfection of miR-10 b mimics in GES-1 cells, the expression of E-cadherin obviously decreased,the expression of Bcl2 effectively increased, and the expression of Bax effectively decreased. The cell proliferation inhibition rates obviously increased, and the difference was statistically significant compared with the control group( P < 0. 05). Conclusion: MiR-10 b can promote the proliferation and suppress the cell apoptosis of gastric cancer cells,and its mechanism may be related to the regulation of E-cadherin.

关键词(KeyWords)： 胃肿瘤;细胞增殖;细胞凋亡;微小RNA10b;上皮型钙黏蛋白
stomach neoplasms;cell proliferation;apoptosis;miR-10b;E-cadherin

Abstract:

Keywords:

基金项目(Foundation): 四川省卫计委项目(17PJ013)

作者(Author): 张雪莲,吴维宇,何峰
ZHANG Xuelian,WU Weiyu,HU Feng

DOI: 10.19367/j.cnki.2096-8388.2020.09.013

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