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

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,West...

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Published inJie fang jun yi xue za zhi Vol. 42; no. 11; pp. 985 - 991
Main Author 车明文;龚翔;张小金;魏达锋;王晓文;王汉民;巨立中
Format Journal Article
LanguageChinese
Published Beijing People's Military Medical Press 2017
841000新疆库尔勒,解放军273医院内三科%845350新疆克州,武警克州支队卫生队%510000广州,空军广州天河干休所%510000广州,空军95269部队53分队保健科%710032西安,空军军医大学西京医院肾脏内科
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ISSN0577-7402
DOI10.11855/j.issn.0577-7402.2017.11.09

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Summary: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(P〈0.05)。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。
Bibliography:Objective To explore the effect of long noncoding RNA-ATB(LncRNA-ATB) on phenotypic transition and proliferation of human peritoneal mesothelial cells(HPMCs) induced by high glucose. Methods HPMCs used in experiment were divided into three groups: control group, mannitol group and hypertonic glucose group. HPMCs in control group received no treatment, and in hypertonic glucose group and mannitol group were treated with 50 mmol/L D-glucose and isotonic mannitol for 72 hours, respectively. Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB, E-cadherin, α-smooth muscle actin(α-SMA), connective tissue growth factor(CTGF), Cyclin D1, cyclin dependent kinase inhibitor 4(CDK4), protein 27(p27)and proliferating cell nuclear antigen(PCNA). Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA, CTGF, Cyclin D1, CDK4, p27 and PCNA, and flow cytometry was used to test the cell cycle. Lentivirus artifice was used to up-or down-regulate the expression of LncRNA-ATB in untr
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ISSN:0577-7402
DOI:10.11855/j.issn.0577-7402.2017.11.09