摘要
目的观察miR-214-3p在胃癌组织中的表达情况,探讨其对胃癌细胞侵袭和迁移能力的影响及其分子机制。方法通过实时荧光定量PCR(qPCR)检测41对胃癌及癌旁正常组织中miR-214-3p的表达及其与肿瘤临床病理特征的关系;将胃癌细胞MGC-803接种于培养皿,每组设3个复孔,分3组:miR-214-3p上调组(转染miR-214-3p模拟物)、miR-214-3p下调组(转染miR-214-3p抑制剂),以及阴性对照(NC)组;qPCR法分别检测各组p53表达水平,Transwell实验检测转染miR-214-3p抑制剂对MGC-803细胞的迁移和侵袭能力的影响;生物信息学分析和荧光素酶活性测定以确定p53是否为miR-214-3p的靶基因。结果相比于癌旁组织,miR-214-3p在胃癌组织中上调,并且与肿瘤淋巴结转移相关;qPCR检测分析显示,相比于NC组,miR-214-3p上调组p53的表达下降,miR-214-3p下调组p53的表达上升;Transwell实验结果显示,与NC组相比,miR-214-3p下调组胃癌细胞的迁移和侵袭能力降低;生物信息学分析和荧光素酶活性测定结果显示miR-214-3p通过靶向结合p533′UTR从而抑制其表达。结论miR-214-3p在胃癌发生发展过程中可能发挥了重要作用,miR-214-3p可作为胃癌的潜在靶向标志物。
Objective To observe the expression of miR-214-3 p in gastric cancer tissues, explore its effect on the metastatic ability of gastric cancer cells, and explore the mechanism of its role. Methods The expression of miR-214-3 p in 41 pairs of gastric cancer and adjacent normal tissues and its relationship with tumor clinicopathological characteristics were detected by Real-time quantitative PCR;gastric cancer cell MGC-803 was inoculated in culture dishs, with 3 replicate wells in each group. The experiment was divided into 3 groups: miR-214-3 p up-regulated group(transfected miR-214-3 p mimic), miR-214-3 p down-regulated group(transfected miR-214-3 p inhibitor), and a NC group was set respectively;Real-time quantitative PCR was used to detect the expression levels of p53 in each group, and the Transwell experiment was used to detect the migration and invasion ability of MGC-803 cells after transfection with miR-214-3 p inhibitor. Bioinformatics analysis and luciferase activity determination determine whether p53 is the target gene of miR-214-3 p. Results Compared with adjacent tissues, miR-214-3 p was up-regulated in gastric cancer tissues and was associated with tumor lymph node metastasis;compared with the NC group, the expression of p53 in the miR-214-3 p up-regulated group decreased, and the expression of p53 increased in the miR-214-3 p down-regulated group;the results of Transwell experiment showed that compared with the NC group, cells of the miR-214-3 p down-regulated group had lower migration and invasion capabilities;bioinformatics analysis and luciferase activity measurement results showed that miR-214-3 p inhibited p53 expression by targeting its 3′UTR. Conclusion miR-214-3 p may play an important role in gastric cancer development and progression, and miR-214-3 p can be used as a potential targeted biomark for gastric cancer.
作者
周大新
谢方利
刘海科
丁鹏
毛作周
孙凯
俞磊
姚卫洲
姜连春
李祥兵
刘聪
吴丽颖
Zhou Daxin;Xie Fangli;Liu Haike(Huaibei Clinical College of Xuzhou Medical University,Huaibei 235100;Huaibei People's Hospital,Dept of General Surgery,Huaibei 235100)
出处
《安徽医科大学学报》
CAS
北大核心
2021年第5期769-773,共5页
Acta Universitatis Medicinalis Anhui
基金
安徽省重点研究和开发计划项目(编号:1804h0820231)
淮北市科技计划项目(编号:rJ201820)。
关键词
胃癌
微小RNA
肿瘤迁移和侵袭
gastric cancer
microRNA
tumor migration and invasion