HOTTIP通过竞争性结合miR-506调节Vimentin基因的表达调控肾透明细胞癌细胞迁移  

HOTTIP regulates the expression of Vimentin gene through competitively binding miR-506 to regulate the migration of clear cell renal cell carcinoma cells

作　　者：王晓玲 孟莉丹 王学敏 张颖轩 张成果 WANG Xiaoling;MENG Lidan;WANG Xuemin;ZHANG Yingxuan;ZHANG Chengguo(Department of Nephrology,People's Hospital of Hengshui,Hebei Hengshui 053000,China)

机构地区：[1]衡水市人民医院肾内科,河北衡水053000

出　　处：《现代肿瘤医学》2024年第7期1200-1207,共8页Journal of Modern Oncology

基　　金：河北省医学科学研究课题计划(编号:20220452)。

摘　　要：目的:探究HOTTIP通过竞争性结合miR-506来调节Vimentin基因的表达调控肾透明细胞癌细胞迁移的作用机制。方法:通过生信分析TCGA数据库中肾透明细胞癌的差异表达miRNA、mRNA、lncRNA。利用miRcode数据库数据对lncRNA-miRNA的结合进行预测,并对miRNA调控的靶基因进行预测。qRT-PCR检测肾透明细胞癌细胞系中HOTTIP、miR-506、Vimentin的表达,RNA pull-down实验检测miR-506和HOTTIP结合,RIP实验检测miR-506与HOTTIP和Vimentin的结合,Transwell和划痕愈合实验检测786-O细胞迁移能力。结果:HOTTIP在肾透明细胞癌中显著上调,并且与患者预后显著相关,其靶miRNA miR-506在肾透明细胞癌中显著下调。miR-506的下游调控基因Vimentin在肾透明细胞癌中显著上调并且与患者预后显著相关。HOTTIP通过竞争性结合miR-506来调节Vimentin基因的表达。干扰HOTTIP后,肾透明细胞癌细胞的迁移能力显著降低,而干扰HOTTIP的同时沉默miR-506或过表达Vimentin则能逆转干扰HOTTIP对肾透明细胞癌细胞迁移能力的抑制作用。结论:HOTTIP很有可能通过竞争性结合miR-506来调节Vimentin基因的表达从而调控肾透明细胞癌细胞迁移。Objective:To explore the mechanism of HOTTIP on regulating the expression of Vimentin gene to regulate the migration of clear cell renal cell carcinoma(ccRCC)cells through competitively binding miR-506.Methods:Bioinformatics analysis was performed on differentially expressed miRNA,mRNA and lncRNA of ccRCC in The Cancer Genome Atlas database.miRcode database was used to predict the binding of lncRNA-miRNA.The target gene regulated by miRNA was also predicted.The expression of HOTTIP,miR-506 and Vimentin mRNA in ccRCC cell lines were detected by qRT-PCR.The binding of miR-506 and HOTTIP was detected by RNA pull-down assay.The binding of miR-506 and HOTTIP or Vimentin was detected by RNA binding protein immunoprecipitation assay.The migratory ability of ccRCC 786-O cells was detected by Transwell and wound healing assays.Results:HOTTIP was significantly upregulated in ccRCC and significantly related to the prognosis of patients.The target miRNA of HOTTIP(miR-506)was markedly downregulated in ccRCC.Downstream regulatory gene Vimentin of miR-506 was significantly upregulated in ccRCC,and significantly related to the prognosis of patients.HOTTIP regulated the expression of Vimentin gene through competitively binding miR-506.Cell migratory ability was significantly decreased after interfering with HOTTIP.However,the inhibitory effect of cell migratory ability by interfering with HOTTIP could be reversed by silencing miR-506 or overexpressing Vimentin simultaneously.Conclusion:HOTTIP could regulate the expression of Vimentin gene to regulate the migration of ccRCC through competitively binding miR-506.

关 键 词：HOTTIP miR-506 VIMENTIN 肾透明细胞癌 迁移 

分 类 号：R737.11[医药卫生—肿瘤]