Prohibitin基因转染对肾小管上皮细胞表型转化的影响  

Effect of Prohibitin Gene on Epithelium Transdifferentiation of Renal Tubular Epithelial Cells Line

作　　者：邵明斌[1] 覃远汉[1] 彭单单[1] 周添标[1] 徐会凌[1] 王勇[1] 雷凤英[1] 李正一[1] 

机构地区：[1]广西医科大学第一附属医院儿科,南宁530021

出　　处：《实用儿科临床杂志》2012年第5期324-326,共3页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(81060061)

摘　　要：目的探讨转染外源性Prohibitin(PHB)基因对体外培养的肾小管上皮细胞株(NRK-52E)表型转化的影响。方法将构建的pcDNA3.1(+)-PHB1、pcDNA3.1(+)-PHB2质粒转染体外培养的NRK-52E细胞作为转染组,以空载体转染NRK-52E细胞为阴性对照组,以正常NRK-52E细胞为空白对照组,分别应用实时荧光定量PCR和Western blot检测转染24 h、48 h、72 h各组NRK-52E细胞PHB1、PHB2及α-平滑肌肌动蛋白(α-SMA)的mRNA及蛋白表达。结果 1.转染24 h、48 h、72 h时转染组NRK-52E细胞的PHB1、PHB2 mRNA及其蛋白表达均较阴性对照组和空白对照组增高(Pa<0.05),且48 h表达量最高,而阴性对照组与空白对照组比较差异无统计学意义。2.转染24 h、48 h、72 h时转染组NRK-52E细胞α-SMA mRNA和蛋白表达均较阴性对照组和空白对照组降低(Pa<0.05),48 h表达量最低,阴性对照组与空白对照组比较差异无统计学意义。3.转染组48 h NRK-52E细胞PHB1、PHB2蛋白表达量与α-SMA蛋白表达量均呈负相关(r=-0.942、-0.869,Pa<0.05)。结论转染外源性PHB基因可以抑制体外培养的NRK-52E细胞的表型转化。Objective To explore the effect of prohibitin gene on epithelium transdifferentiation of renal tubular epithelial cells line（NRK-52E）. Methods Cultured NRK-52E cells were transfected with pcDNA3.1（＋）-PHB1,pcDNA3.1（＋）-PHB2 plasmid（transfection group） or empty carrier（negative control group）,with normal NRK-52E cells as blank control group.The gene and protein expressions of PHB1,PHB2,and α-smooth muscle actin（α-SMA）were detected by using real-time fluorescent quantitive polymerase chain reaction（PCR） and Western blot in 24 hours,48 hours,and 72 hours after transfection. Results 1.In 24 hours,48 hours,and 72 hours after transfection,the mRNA and protein expressions of PHB1 and PHB2 were higher in the transfection group than those in the negative control group and blank control group（Pa0.05）.Those expressions in 48 h were higher when compared with those in other groups,and the diffe-rence between negative control group and blank control group was not statistically significant.2.The mRNA and protein expressions of α-SMA were lower in the transfection group of PHB1 and PHB2 than those in the negative control group and blank control group（Pa0.05）,and those expressions in 48 hours were the lowest and the difference between negative control group and blank control group was not statistically significant.3.Correlation analysis showed that：The expressions of protein of PHB1 and PHB2 in transfection group were negatively correlated with the protein expression of α-SMA（r=-0.942,-0.869,Pa0.05）. Conclusion The transfection using PHB mediated by liposomes can inhibit renal tubular epithelial cells from phenotypic transformation.

关 键 词：PROHIBITIN 肾小管上皮细胞 转染 Α-平滑肌肌动蛋白 表型转化 

分 类 号：R726.91[医药卫生—儿科]