Twist过表达在人腹膜间皮细胞转分化中的作用  

Effects of twist over-expression on the transdifferentiation of human peritoneal mesothelial cells

作　　者：李翠香[1] 孙世仁[1] 何丽洁[1] 杜锐[1] 娄未娟[1] 王汉民[1] 

机构地区：[1]第四军医大学西京医院肾脏内科,西安710032

出　　处：《解放军医学杂志》2011年第5期456-458,462,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81070570)

摘　　要：目的探讨twist在人腹膜间皮细胞(HPMCs)转分化中的作用。方法取HPMCs细胞,分别用50mmol/L右旋葡萄糖(高糖组)和5.6mmol/L右旋葡萄糖(对照组)刺激48h,采用Western blotting检测twist、E-钙黏素(E-cadherin)、成纤维细胞特异性蛋白1(Fsp1)和α-平滑肌肌动蛋白(α-SMA)的表达情况。另采用质粒pcDNA3.1-twist(pcDNA3.1-twist组)和空载体pcDNA3.1(空载体组)分别转染常规培养的HPMCs,使HPMCs过表达twist后,采用qRT-PCR、Western blotting方法检测twist、E-cadherin、Fsp1和α-SMA mRNA及蛋白的表达情况。再采用质粒twist小干扰RNA(阳性转染组)和空载体pcDNA3.1(空载体组)分别转染50mmol/LD-葡萄糖培养48h后的HPMCs细胞,另选择未转染质粒的细胞作为亲本细胞组,采用Western blotting检测twist下调后,E-cadher-in、Fsp1、α-SMA蛋白水平的表达情况。结果与对照组相比,高糖组twist、Fsp1和α-SMA蛋白表达明显增加,E-cadherin表达减弱(P<0.05)。与空载体组相比,HPMCs过表达twist后,pcDNA3.1-twist组E-cadherin mRNA及蛋白表达减弱,Fsp1和α-SMA mRNA及蛋白表达增强(P<0.05)。用小干扰RNA使twist沉默后,与亲本细胞组及空载体相比,E-cadherin蛋白表达增加,Fsp1和α-SMA蛋白表达减弱(P<0.05)。结论 Twist过表达促进了HPMCs的转分化。Objective To explore the effects of twist over-expression on the transdifferentiation of human peritoneal mesothelial cells（HPMCs）.Methods The HPMCs were treated separately with 50mmol/L（high glucose group） and 5.6mmol/L（control group） D-glucose for 48h.The protein expression of twist,E-cadherin,Fsp1 and α-SMA were detected by Western blotting.The mRNA and protein expression of twist,E-cadherin,Fsp1 and α-SMA were tested by qRT-PCR and Western blotting after transfecting the plasmids pcDNA3.1-twist and empty vector pcDNA3.1 into HPMCs with Lipofectamine 2000 in vitro.The protein expression of twist,E-cadherin,Fsp1 and α-SMA were tested by Western blotting after silencing the twist expression by siRNA,empty vector pSlience and the parental cells.Results Compared with control group,the expression of twist,Fsp1 and α-SMA increased,and of E-cadherin decreased in high glucose group（P〈0.05）.Compared with the empty vector group,after the over-expression of twist in HPMCs,the expression of E-cadherin decreased,and of Fsp1 and α-SMA increased in high glucose group（P〈0.05）.Silencing the twist expression by siRNA,the expression of E-cadherin increased and of α-SMA and Fsp1 decreased in siTwist transfacted cells compared with pSlicencer-transfacted cell and parental cells（P〈0.05）,and finally reversed the phenotype of MMT.Conclusion The over-expression of twist may promote the mesothelial-to-mesenchymal transition of HPMCs.

关 键 词：间皮细胞间质转分化 TWIST转录因子 钙黏着糖蛋白类 肌动蛋白类 

分 类 号：R459.5[医药卫生—治疗学] R329.24[医药卫生—临床医学]