LncRNA-ATB在高糖诱导的人腹膜间皮细胞表型转换及增殖中的作用

发布时间：2018-03-27 19:48

本文选题：细胞表型转换　切入点：LncRNA-ATB　出处：《解放军医学杂志》2017年11期

【摘要】：目的探讨长链非编码RNA-ATB(Lnc RNA-ATB)在高糖刺激诱导的人腹膜间皮细胞(HPMCs)表型转换及增殖中的作用。方法将HPMCs分为对照组、甘露醇组和高糖组3组。对照组不予任何处理;高糖组和甘露醇组分别采用50mmol/L D型葡萄糖和同等渗透压的甘露醇刺激72h。采用Real-time PCR检测各组Lnc RNA-ATB、E-钙黏素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、结缔组织生长因子(CTGF)、细胞周期素D1(Cyclin D1)、细胞周期蛋白依赖激酶抑制因子4(CDK4)、蛋白质p27(p27)及增殖细胞核抗原(PCNA)m RNA的表达,Western blotting检测各组E-cadherin、α-SMA、CTGF、Cyclin D1、CDK4、p27及PCNA蛋白的表达,并采用流式细胞仪检测细胞周期。利用慢病毒转染技术,将Lenti-Lnc RNA-ATB和sh RNA-Lnc RNA-ATB导入HPMCs中,分别上调及下调Lnc RNA-ATB的表达,采用Real-time PCR检测E-cadherin、α-SMA及CTGF m RNA的表达,Western blotting检测E-cadherin、α-SMA、CTGF蛋白的表达,流式细胞仪检测细胞周期。结果 Real-time PCR、Western blotting及流式细胞仪检测结果显示,高糖刺激后,Lnc RNA-ATB、α-SMA、CTGF、Cyclin D1、CDK4及PCNA表达增加,E-cadherin及p27表达减少(P0.05)。上调Lnc RNAATB表达可促进细胞表型转换及增殖,下调Lnc RNA-ATB表达可抑制细胞表型转换及增殖。结论高糖刺激可通过上调Lnc RNA-ATB的表达促进人腹膜间皮细胞表型转换及增殖。
[Abstract]:Objective to investigate the role of long chain noncoding RNA-ATB(Lnc RNA-ATB in the phenotypic transformation and proliferation of human peritoneal mesothelial cells stimulated by high glucose. Methods HPMCs was divided into three groups: control group, mannitol group and high glucose group. The high glucose group and mannitol group were stimulated with 50mmol/L D glucose and mannitol for 72 h, respectively. The Lnc RNA-ATBU E-cadherin, 伪 -SMAG (伪 -SMAN), D1(Cyclin D1 (cyclin D 1), D1(Cyclin D 1 (Cyclin D 1) were detected by Real-time PCR in each group. The expression of cyclin dependent kinase inhibitor (p27p27) and proliferating cell nuclear antigen (PCNA RNA) was detected by Western blotting to detect the expression of E-cadherin, 伪 -SMA-CTGFGFCF-Cyclin D1, CDK4p27 and PCNA protein in each group. The cell cycle was detected by flow cytometry. Lenti-Lnc RNA-ATB and sh RNA-Lnc RNA-ATB were transfected into HPMCs with lentivirus transfection technique. The expression of Lnc RNA-ATB was up-regulated and down-regulated, respectively. E-cadherin, 伪 -SMA and CTGF m RNA were detected by Real-time PCR and E-cadherin and 伪 -SMACTGF were detected by Western blotting. Results Real-time PCR Western blotting and flow cytometry showed that the expression of Lnc RNA-ATB, 伪 -SMA-CTGFGFCyclin D1 CDK4 and PCNA increased after high glucose stimulation, and the expression of E-cadherin and p27 decreased. Upregulating the expression of Lnc RNAATB could promote the phenotypic transformation and proliferation of the cells. Conclusion High glucose stimulation can promote the phenotypic transformation and proliferation of human peritoneal mesothelial cells by up-regulating the expression of Lnc RNA-ATB.
【作者单位】：解放军273医院内三科;武警克州支队卫生队;空军广州天河干休所;空军95269部队53分队保健科;空军军医大学西京医院肾脏内科;
【分类号】：R459.5

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