摘要
目的了解hsa-miR-653-5p在胃癌中的表达水平及与胃癌临床病理特征的关系,初步探讨hsa-miR-653-5p在胃癌进展中的功能作用。方法本文利用TCGA数据库的数据评估得出hsa-miR-653-5p与胃癌患者预后显著相关,并分析hsamiR-653-5p在胃腺癌组织和正常胃组织中的表达水平;同时评估胃癌组织中hsa-miR-653-5p的表达水平与胃癌患者的淋巴结个数及临床预后的临床特征关系,利用TCGA数据库和KEGG通路富集分析预测hsa-miR-653-5p的潜在分子生物学功能,构建hsa-miR-653-5p过表达及低表达细胞株行transwell实验验证hsa-miR-653-5p在胃癌细胞中的生物学功能;根据miRNA靶基因数据库及TCGA数据库来预测和筛选hsa-miR-653-5p的靶基因,通过Western blot检测转染前后胃癌细胞内PTEN蛋白表达水平的变化,并采用双荧光素酶报告基因实验验证hsa-miR-653-5p与靶基因PTEN的相互作用。结果 hsa-miR-653-5p在胃腺癌组织中的表达水平显著高于正常胃组织,hsa-miR-653-5p高表达的胃腺癌患者具有更差的生存率,hsa-miR-653-5p的表达水平与淋巴结数量呈正相关;通路富集分析表明hsa-miR-653-5p可能参与了粘附连接、粘着斑、胃癌、磷脂酰肌醇信号系统的生物过程和途径;transwell实验证实hsa-miR-653-5p促进胃癌细胞BGC823和SGC7901的迁移;hsa-miR-653-5p直接靶向并负调控PTEN促进胃癌的发生。结论 hsa-miR-653-5p在胃腺癌组织中表达上调并与胃癌患者预后呈负相关,hsa-miR-653-5p可能通过负调控抑癌基因PTEN促进胃癌细胞的迁移。
Objective The aim of this study is to identify whether hsa-miR-653-5 p plays a role in the development of gastric cancer( GC). Methods This article used the TCGA database to evaluate that hsa-miR-653-5 p was significantly correlated with the prognosis of patients with gastric cancer. The expression of hsa-miR-653-5 p in gastric adenocarcinoma and normal gastric tissues was obtained from the TCGA database. We also assessed the association of hsa-miR-653-5 p expression with GC clinicopathological features and investigated the impact of hsa-miR-653-5 p on GC survival. Potential molecular function of hsa-miR-653-5 p was predicted through TCGA database and functional enrichment. Migration assay was used to validate the biological function of hsa-miR-653-5 p in GC. Target genes of hsa-miR-653-5 p were predicted online using the TCGA database and miRNA target gene database. Western blot was performed to validate the negative correlation relationship between hsa-miR-653-5 p and PTEN and a dual-luciferase reporter assay was designed to confirm that PTEN was the target gene of hsa-miR-653-5 p. Results Compared with normal gastric tissues,the expression of hsa-miR-653-5 p was significantly higher in stomach adenocarcinoma tissues. Patients with stomach adenocarcinoma with high expression of hsamiR-653-5 p had a worse survival. Hsa-miR-653-5 p expression was positively correlated with the number of lymph nodes. KEGG Pathway analysis indicated that hsa-miR-653-5 p might be involved in Adherens junction,Focal adhesion,Gastric cancer,Phosphatidylinositol signaling system. Hsa-miR-653-5 p promoted the migration of gastric cancer cells. By using luciferase reporter assay,we found PTEN was a direct downstream target of hsa-miR-653-5 p,and negatively regulated by hsa-miR-653-5 p via targeting its 3’ UTR. Knockdown or overexpression of hsa-miR-653-5 p in BGC823 cells negatively regulated PTEN expression. Conclusion Hsa-miR-653-5 p is upregulated in gastric adenocarcinoma and negatively correlated with the prognosis of gastric canc
作者
禤文真
何杰
杜艳蕾
赵冲
徐豪明
聂玉强
XUAN Wen-zhen;HE Jie;DU Yan-lei;ZHAO Chong;XU Hao-ming;NIE Yu-qiang(Department of Gastroenterology,Guangzhou First People′s Hospital,Guangzhou Medical University,Department of Gastroenterology,Guangzhou First People′s Hospital,School of medicine,South China University of Technology,Guangzhou,Guangdong,China,510180)
出处
《现代消化及介入诊疗》
2019年第12期1366-1373,共8页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
国家自然科学基金(81871905)
广州市科创委项目(201707010275)
