Yang Peng, Zhang Mei, Liu Xiting, Pu Huayun
Department of Nuclear Medicine, The First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
Department of Gastroenterology, General Hospital of Lanzhou Petrochemical Company, Lanzhou, Gansu 730060, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2625-2632. doi: 10.3892/etm.2017.4798. Epub 2017 Jul 17.
The present study aimed to evaluate the expression of microRNA (miR)-421 in gastric cancer and to investigate its biological function and underlying mechanism of action in the development of gastric cancer. The expression of miR-421 was measured in 60 pairs of clinically removed gastric cancer tissues and matched adjacent normal gastric tissues by reverse transcription-quantitative polymerase chain reaction. In addition, following transfection with an miR-421 inhibitor to suppress the expression of miR-421, the proliferation, migration and cell cycle distribution of human gastric carcinoma MKN28/MKN74 cells were determined by cell counting, Transwell and flow cytometry assays. The target gene of miR-421 was also predicted using bioinformatic analysis and verified by dual-luciferase reporter gene assay and western blot analysis. Furthermore, overexpression of the miR-421 target protein was induced in MKN28/MKN74 cells to determine its function. It was observed that miR-421 was significantly upregulated in gastric cancer tissues and that the expression of miR-421 was associated with lymph node metastasis and the clinical stage of gastric cancer (all P<0.05). Claudin11 (CLDN11) was predicted and verified as a direct target of miR-421. experiments demonstrated that inhibition of miR-421 expression suppressed the proliferation and metastasis of MKN28/MKN74 cells and induced G1/S-phase cell cycle arrest (all P<0.05). Analagous results were observed in MKN28/MKN74 cells following overexpression of the CLDN11 protein. Collectively, these data suggest that miR-421 may promote the proliferation, invasion and metastasis of gastric cancer by inhibiting the expression of CLDN11.
本研究旨在评估微小RNA(miR)-421在胃癌中的表达,并探讨其在胃癌发生发展中的生物学功能及潜在作用机制。采用逆转录-定量聚合酶链反应检测60对临床切除的胃癌组织及其配对的癌旁正常胃组织中miR-421的表达。此外,转染miR-421抑制剂以抑制miR-421的表达后,通过细胞计数、Transwell和流式细胞术检测人胃癌MKN28/MKN74细胞的增殖、迁移及细胞周期分布。还利用生物信息学分析预测miR-421的靶基因,并通过双荧光素酶报告基因检测和蛋白质印迹分析进行验证。此外,在MKN28/MKN74细胞中诱导miR-421靶蛋白的过表达以确定其功能。结果显示,miR-421在胃癌组织中显著上调,且miR-421的表达与胃癌的淋巴结转移及临床分期相关(均P<0.05)。Claudin11(CLDN11)被预测并验证为miR-421的直接靶标。实验表明,抑制miR-421表达可抑制MKN28/MKN74细胞的增殖和转移,并诱导G1/S期细胞周期阻滞(均P<0.05)。在CLDN11蛋白过表达的MKN28/MKN74细胞中观察到类似结果。综上所述,这些数据表明miR-421可能通过抑制CLDN11的表达促进胃癌的增殖、侵袭和转移。