摘要
目的:探索胃癌细胞中高表达hnRNP K蛋白直接调控的miRNA。方法:通过制备正常胃黏膜细胞GES-1和胃癌细胞SGC-7901的细胞芯片初筛高表达hnRNP K胃癌细胞miRNA表达谱;查询TCGA数据库得到胃癌大样本mRNA转录组和miRNA数据,分析处理得到hnRNP K以2倍差异高表达对应的miRNA差异表达谱;对芯片和TCGA数据库结果交互分析得到上调共表达的miRNA;过表达和敲除hnRNP K,通过Western Blot和Real Time q-PCR验证筛查的结果;利用TCGA大数据对hnRNP K和miRNA进行归一化处理,验证筛选的miRNA与hnRNP K和胃癌的相关性。结果:通过芯片初筛miRNA表达谱与TCGA查询结果交互分析得到胃癌上调miRNA共表达谱(miR-17/20a/24/182);Western Blot和Real Time q-PCR验证结果显示hnRNP K可直接调控miR-20a的表达;TCGA数据分析表明miR-20a与hnRNP K正相关,且在胃癌中高表达。结论:胃癌细胞中高表达hnRNP K蛋白上调miR-20a的表达。
Objective:To explore the expression of microRNAs directly regulated by hnRNP K protein in gastric cancer cells.Methods:The microRNA expression profiles of gastric cancer cells with high expression of hnRNP K were screened by cell microarray of normal gastric mucous membrane cell GES-1 and gastric cancer cell line SGC-7901.Large samples of transcription group and microRNA data were obtained by querying TCGA database,and the microRNA differential expression profiles corresponding to hnRNP K with high expression of 2-fold difference were obtained by analysis and processing.The co-expression of microRNAs was up-regulated by interactive analysis of the results of microarray and TCGA database.hnRNP K was overexpressed or knocked down and the results of screening were validated by Western Blot and Real Time q-PCR.hnRNP K and microRNAs were normalized with TCGA data to verify the correlation between the screened microRNAs and hnRNP K in gastric cancer.Results:The co-expression profile of miR-17/20a/24/182 was up-regulated by microarray screening and TCGA query results.Western Blot and Real Time q-PCR results showed that hnRNP K could directly regulate the expression of miR-20a.TCGA data analysis showed that miR-20a was positively correlated with hnRNP K and highly expressed in gastric cancer.Conclusion:High expression of HnRNP K protein in gastric cancer cells up-regulates the expression of miR-20a.
作者
方海云
郭佳慧
高丰厚
Fang Haiyun;Guo Jiahui;Gao Fenghou(Department of Oncology,Shanghai Ninth People's Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 201900,China;Department of Laboratory Medicine,Xinhua Hospital Chongming Branch,Shanghai Jiaotong University School of Medicine,Shanghai 202150,China)
出处
《现代肿瘤医学》
CAS
2020年第5期706-712,共7页
Journal of Modern Oncology
基金
国家自然科学基金(编号:81172322)
