草酸钠结晶肾损伤肾小管上皮细胞中骨桥蛋白相关长链非编码RNA的差异表达  

Differential expression of osteopontin-related long non-coding RNAs in sodium oxalate crystallization-induced renal tubular epithelial cell injury

作　　者：张杰[1] 谌卫[1] 袁继行 王欢[1] 马金召 王田田[2] 郭志勇[1] 

机构地区：[1]第二军医大学长海医院肾内科,上海200433 [2]第二军医大学基础部医学遗传学研究所,上海200433

出　　处：《第二军医大学学报》2017年第5期616-621,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81270773;81573759);上海市科委中药支撑计划(13401900105);上海市教委科研创新重点项目(14ZZ080)~~

摘　　要：目的检测草酸钠诱导的人肾小管上皮细胞结晶肾损伤模型中与骨桥蛋白(OPN)相关的长链非编码RNA(lncRNA)的差异表达,并探讨OPN相关lncRNA在结晶肾损伤发病中的作用。方法利用草酸钠(20 mmol/L)刺激人肾近曲小管上皮细胞HK-2建立草酸钠诱导的结晶肾损伤肾小管上皮细胞模型。运用RNA干扰技术对其转染OPN小干扰RNA构建OPN敲低细胞模型(OPN-siRNA组),对照组细胞转染阴性随机对照序列(NC-siRNA组)。采用Arraystar公司提供的人类全基因组lncRNA芯片(V4.0)检测两组结晶肾损伤肾小管上皮细胞模型中与OPN相关的lncRNA和mRNA的差异表达水平,并采用生物信息学方法分析lncRNA芯片结果。结果全基因组lncRNA芯片结果显示草酸钠诱导的结晶肾损伤肾小管上皮细胞模型中与OPN相关的差异表达lncRNA共583个,其中OPN-siRNA组表达上调354个、下调229个;差异表达mRNA共235个,上调139个,下调96个。生物信息学分析结果显示差异表达lncRNA参与结晶肾损伤中细胞生长、酶活性调节等多种生物学过程。结论草酸钠体外诱导的结晶肾损伤肾小管上皮细胞模型中存在OPN相关的差异表达的lncRNA。Objective To detect the differential expression of osteopontin （OPN）-related long non-coding RNAs （lncRNAs） in sodium oxalate crystallization-induced renal tubular epithelial cell injury and to explore the role of OPN-related lncRNAs in the pathogenesis of renal injury by crystallization. Methods The renal tubular epithelial cells （HK-2 cells） were cultured and stimulated with 20 mmol/L sodium oxalate to establish the renal injury cells model. The OPN knockdown in HK-2 cell injury was carried out by transfecting OPN small interfere RNA （siRNA） into the cells, which was set up as the OPN-siRNA group; the cells in control group （NC-siRNA group） were transfected with negative random control sequences. The lncRNAs and mRNAs differential expressions were screened and identified by Arraystar Human lncRNA V4.0 Microarray in two groups, and the microarray results were analyzed by bioinformatics methods. Results Human genome lncRNA microarray results showed there were a total of 583 differentially expressed OPN-related lncRNAs in renal injury cell model of renal tubular epithelial cell induced by sodium oxalate crystallization, including 354 up-regulated lncRNAs and 229 down-regulated lncRNAs in OPN-siRNA group; there were a total of 235 mRNAs with differential expression, including 139 up-regulated mRNAs and 96 down-regulated mRNAs. Bioinformatics analysis results showed the differentially expressed lncRNAs were correlated with cell growth, enzyme activity regulation and other biological processes. Conclusion OPN-related differentially expressed lncRNAs are involved in the mechanism of the sodium oxalate crystallization-induced HK-2 cell injury.

关 键 词：肾结石 结晶肾损伤 肾小管上皮细胞 草酸钠 骨桥蛋白 长链非编码RNA 

分 类 号：R692.4[医药卫生—泌尿科学]