Department of Clinical Laboratory, The First People's Hospital of Changzhou, Changzhou, Jiangsu, Chile.
Department of Clinical Laboratory, The First People's Hospital of Changzhou, Changzhou, Jiangsu, China (mainland).
Med Sci Monit. 2019 Aug 24;25:6367-6377. doi: 10.12659/MSM.915970.
BACKGROUND This study aimed to investigate the role of micro-RNA 205-5p (miR-205-5p) in the progression of gastric cancer, and the target of miR-205-5p in human gastric cancer cells in vitro. MATERIAL AND METHODS Expression of miR-205-5p and PTEN in gastric cancer tissue samples and adjacent normal gastric tissue from 35 patients was studied using immunohistochemistry and in situ hybridization. SGC-7901 human gastric cancer cells included a normal control (NC) group, a group transfected with empty vector (Vector), a group treated with miR-205-5p inhibitor (miR-inhibitor), and a group treated with miR-205-5p inhibitor and small interfering PTEN mRNA (miR-inhibitor+si-PTEN). Quantitative reverse transcription polymerase chain reaction (qRT-PCR) measured miR-205-5p expression, cell proliferation was measured by MTT assay, cell apoptosis by flow cytometry, transwell and wound healing assays measured cell migration, and transmission electron microscopy (TEM) showed ultrastructural changes in SGC-7901 cells. PTEN, AKT and p-AKT protein expression were measured using Western blot. The correlation between miR-205-5p and PTEN was analyzed using a dual-luciferase reporter assay. RESULTS Increased expression of miR-205-5p and PTEN in gastric cancer tissues were correlated with tumor stage. In SGC-7901 cells, miR-205-5p mRNA expression in the miR-inhibitor and miR-inhibitor+si-PTEN groups was significantly lower than that in the NC group (P<0.001). In the miR-inhibitor group, cell proliferation was significantly decreased, and apoptosis was significantly increased (P<0.001). CONCLUSIONS In gastric cancer, increased expression of miR-205-5p was associated with tumor stage, and in SGC-7901 cells PTEN was a target gene for miR-205-5p.
本研究旨在探讨微小 RNA 205-5p（miR-205-5p）在胃癌进展中的作用，以及 miR-205-5p 在体外人胃癌细胞中的靶标。
采用免疫组织化学和原位杂交技术检测 35 例胃癌组织标本及癌旁正常胃组织中 miR-205-5p 和 PTEN 的表达。人胃癌 SGC-7901 细胞分为正常对照组（NC 组）、空载体转染组（Vector 组）、miR-205-5p 抑制剂转染组（miR-inhibitor 组）和 miR-205-5p 抑制剂联合小干扰 PTEN mRNA 转染组（miR-inhibitor+si-PTEN 组）。采用实时荧光定量聚合酶链反应（qRT-PCR）检测 miR-205-5p 表达，MTT 法检测细胞增殖，流式细胞术检测细胞凋亡，Transwell 小室和划痕愈合实验检测细胞迁移，透射电镜（TEM）观察 SGC-7901 细胞超微结构变化，Western blot 检测 PTEN、AKT 和 p-AKT 蛋白表达。采用双荧光素酶报告基因检测 miR-205-5p 与 PTEN 的相关性。
胃癌组织中 miR-205-5p 和 PTEN 的表达增加与肿瘤分期相关。在 SGC-7901 细胞中，miR-inhibitor 组和 miR-inhibitor+si-PTEN 组 miR-205-5p mRNA 表达均明显低于 NC 组（P<0.001）。miR-inhibitor 组细胞增殖明显减少，凋亡明显增加（P<0.001）。
在胃癌中，miR-205-5p 的表达增加与肿瘤分期有关，在 SGC-7901 细胞中，PTEN 是 miR-205-5p 的靶基因。
