长链非编码RNA OIP5-AS1在肾癌细胞中的表达及其与细胞增殖和转移的关系  

Expression of long non-coding RNA OIP5-AS1 in renal carcinoma cells and its relationship with cell proliferation and metastasis

作　　者：杨栋 赵鹏程 田沛 马彦 何朝红 Yang Dong;Zhao Pengcheng;Tian Pei;Ma Yan;He Chaohong(Department of Urology,Affiliated Cancer Hospital of Zhengzhou University,Zhengzhou 450008,China)

机构地区：[1]郑州大学附属肿瘤医院泌尿外科,郑州450008

出　　处：《中华实验外科杂志》2024年第6期1148-1151,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划项目(LHGJ20220192)。

摘　　要：目的探讨长链非编码RNA(lncRNA)OIP5-AS1在肾癌细胞中的表达及其与细胞增殖和转移的关系。方法人肾小管上皮细胞系(HK2)和肾癌细胞系ACHN采用转录组学分析肾小管细胞和肾癌细胞差异表达lncRNA。选择表达差异最为显著的lncRNA作为研究对象,采用荧光定量聚合酶链反应(PCR)分析正常肾上皮细胞和肾癌细胞系(ACHN、GRC-1和786-0)lncRNA OIP5-AS1表达水平。肾癌细胞系ACHN分为lncRNA对照组和lncRNA OIP5-AS1 KD组,采用慢病毒建立稳定转染细胞系,细胞计数试剂盒(CCK-8)和肿瘤形成实验分析两组细胞的增殖能力;流式细胞术分析细胞凋亡和细胞周期变化;Transwell分析两组细胞的迁移和侵袭能力;蛋白质免疫印迹分析两组细胞增殖、侵袭和上皮-间充质转化相关蛋白表达水平。组间计量数据比较采用t检验。结果转录组学发现表达最为差异的基因是lncRNA OIP5-AS1。人肾小管上皮细胞系(HK2)lncRNA OIP5-AS1表达水平(0.97±0.13)明显低于肾癌细胞系ACHN、GRC-1和786-0(2.12±0.16、1.68±0.12、1.46±0.11),差异有统计学意义(t=13.870、9.791、7.125,P<0.05)。lncRNA对照组细胞吸光度值、裸鼠体内肿瘤体积和质量[1.99±0.10、(754.50±71.61)cm^(3)、(4.96±0.87)g]明显高于lncRNA OIP5-AS1 KD组细胞[1.55±0.12、(419.17±87.59)cm^(3)、(2.96±0.27)g],差异有统计学意义(t=6.934、5.384、7.260,P<0.05)。lncRNA对照组迁移和侵袭细胞数量[(145.00±9.75)、(89.17±5.38)个]明显高于lncRNA OIP5-AS1 KD组细胞[(110.67±11.52)、(63.17±10.61)个],差异有统计学意义(t=5.571、5.353,P<0.05)。lncRNA对照组细胞核增殖抗原(Ki-67)、黏着斑激酶(FAK)、间充质细胞标志物神经钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达水平(1.02±0.11、1.44±0.16、1.21±0.17、1.44±0.16)明显高于lncRNA OIP5-AS1 KD组细胞(0.50±0.09、0.91±0.09、0.67±0.17、0.69±0.09),差异有统计学意义(t=8.755、7.117、6.509、10.100,P<0.05)。lncRNA对照组�Objective To investigate the expression of long non-coding RNA(lncRNA)OIP5-AS1 in renal carcinoma cells and its relationship with cell proliferation and metastasis.Methods The differential expression of lncRNA in human tubular epithelial cell line(HK2)and renal carcinoma cell line(ACHN)was analyzed by transcriptomics.LncRNA with the most significant expression difference was selected as the study object,and the expression level of lncRNA OIP5-AS1 in normal renal epithelial cells and renal cancer cell lines(ACHN,GRC-1 and 786-0)was analyzed by fluorescence quantitative polymerase chain reaction(PCR).Renal cancer Cell lines(ACHN)were divided into lncRNA control group and lncRNA OIP5-AS1 KD group.Lentivirus was used to establish stable transfected cell lines.Cell proliferation capacity of the two groups was analyzed by cell counting kit-8(CCK-8)and tumor formation assays.Cell cycle was analyzed by flow cytometry.The migration and invasion ability of the two groups of cells was analyzed by Transwell.The expression levels of proteins related to cell proliferation,invasion and epithelial mesenchymal transformation were detected by Western blotting.T test was used to compare measurement data between groups.Results The most differentially expressed gene was lncRNA OIP5-AS1.The expression level of OIP5-AS1 in human renal tubular epithelial cell line(HK2)was significantly lower(0.97±0.13)than that in renal carcinoma cell lines(ACHN,GRC-1 and 786-0)(2.12±0.16,1.68±0.12,1.46±0.11;t=13.870,9.791,7.125;P<0.05).The absorbance value of lncRNA control group,tumor volume and mass in nude mice[1.99±0.10,(754.50±71.61)cm^(3),(4.96±0.87)g]was significantly greater than those of lncRNA OIP5-AS1 KD group[1.55±0.12,(419.17±87.59)cm^(3),(2.96±0.27)g;t=6.934,5.384,7.260;P<0.05].The number of migratory and invasive cells(145.00±9.75,89.17±5.38)in lncRNA control group was significantly greater than that in lncRNA OIP5-AS1 KD group(110.67±11.52,63.17±10.61;t=5.571,5.353,P<0.05).The protein expression levels of proliferating cel

关 键 词：长链非编码RNA 肾癌细胞 增殖 转移 

分 类 号：R737.11[医药卫生—肿瘤]