华中科技大学学报（医学版）
華中科技大學學報（醫學版）
화중과기대학학보（의학판）
ACTA UNIVERSITATIS MEDICINAE TONGJI
2009年
6期
721-724
,共4页
位红兰%曾锐%刘林%张娟%罗昀%葛树旺%徐钢
位紅蘭%曾銳%劉林%張娟%囉昀%葛樹旺%徐鋼
위홍란%증예%류림%장연%라윤%갈수왕%서강
PTEN%肾小管上皮细胞%细胞转分化%PI3K/Akt
PTEN%腎小管上皮細胞%細胞轉分化%PI3K/Akt
PTEN%신소관상피세포%세포전분화%PI3K/Akt
PTEN%renal tubular epithelia%epithelial-mesenchymal trans-differentiation%PI3K/Akt
目的 研究PTEN高表达对TGF-β1诱导肾小管上皮细胞转分化的抑制及其信号传导机制.方法 脂质体介导含人全长PTEN基因或空载体转染人肾小管上皮细胞(HKC细胞)48 h,倒置荧光显微镜检测PTEN转染组及空载体转染组绿色荧光蛋白表达;Western blot检测正常组、PTEN转染组、空载体转染组中PTEN蛋白表达;RT-PCR检测3组中PTEN mRNA表达.然后将实验分为正常组、TGF-β1刺激组(给予TGF-β1刺激)、PTEN+TGF-β1组(PTEN基因转染后给予TGF-β1刺激)、空载体+TGF-β1组(空载体转染后给予TGF-β1刺激),Western blot检测各组细胞E-cad-herin、α-SMA、Akt、p-Akt表达水平.结果 FTEN基因及空载体转染HKC细胞48 h后可见明显绿色荧光蛋白表达;PTEN转染组PTEN蛋白及PTEN mRNA表达较正常组及空载体转染组细胞均显著上升(均P<0.05).在正常组、TGF-β1刺激组、PTEN+TGF-β1组、空载体+TGF-β1组中,TGF-β1刺激组及空载体+TGF-β1组较正常组E-cadherin表达明显下降(均P<0.05),而a-SMA及p-Akt表达显著上升(均P<0.05);PTEN+TGF-β1组较TGF-β1刺激组及空载体+TGF-β1组a-SMA及p-Akt表达明显下降(均P<0.05).而E-cadherin表达上升(均P<0.05);各组细胞Akt表达水平差异不明显(P>0.05).结论 PTEN通过阻止PI3K/Akt通路活化而抑制TGF-β1诱导的肾小管上皮细胞转分化.
目的 研究PTEN高錶達對TGF-β1誘導腎小管上皮細胞轉分化的抑製及其信號傳導機製.方法 脂質體介導含人全長PTEN基因或空載體轉染人腎小管上皮細胞(HKC細胞)48 h,倒置熒光顯微鏡檢測PTEN轉染組及空載體轉染組綠色熒光蛋白錶達;Western blot檢測正常組、PTEN轉染組、空載體轉染組中PTEN蛋白錶達;RT-PCR檢測3組中PTEN mRNA錶達.然後將實驗分為正常組、TGF-β1刺激組(給予TGF-β1刺激)、PTEN+TGF-β1組(PTEN基因轉染後給予TGF-β1刺激)、空載體+TGF-β1組(空載體轉染後給予TGF-β1刺激),Western blot檢測各組細胞E-cad-herin、α-SMA、Akt、p-Akt錶達水平.結果 FTEN基因及空載體轉染HKC細胞48 h後可見明顯綠色熒光蛋白錶達;PTEN轉染組PTEN蛋白及PTEN mRNA錶達較正常組及空載體轉染組細胞均顯著上升(均P<0.05).在正常組、TGF-β1刺激組、PTEN+TGF-β1組、空載體+TGF-β1組中,TGF-β1刺激組及空載體+TGF-β1組較正常組E-cadherin錶達明顯下降(均P<0.05),而a-SMA及p-Akt錶達顯著上升(均P<0.05);PTEN+TGF-β1組較TGF-β1刺激組及空載體+TGF-β1組a-SMA及p-Akt錶達明顯下降(均P<0.05).而E-cadherin錶達上升(均P<0.05);各組細胞Akt錶達水平差異不明顯(P>0.05).結論 PTEN通過阻止PI3K/Akt通路活化而抑製TGF-β1誘導的腎小管上皮細胞轉分化.
목적 연구PTEN고표체대TGF-β1유도신소관상피세포전분화적억제급기신호전도궤제.방법 지질체개도함인전장PTEN기인혹공재체전염인신소관상피세포(HKC세포)48 h,도치형광현미경검측PTEN전염조급공재체전염조록색형광단백표체;Western blot검측정상조、PTEN전염조、공재체전염조중PTEN단백표체;RT-PCR검측3조중PTEN mRNA표체.연후장실험분위정상조、TGF-β1자격조(급여TGF-β1자격)、PTEN+TGF-β1조(PTEN기인전염후급여TGF-β1자격)、공재체+TGF-β1조(공재체전염후급여TGF-β1자격),Western blot검측각조세포E-cad-herin、α-SMA、Akt、p-Akt표체수평.결과 FTEN기인급공재체전염HKC세포48 h후가견명현록색형광단백표체;PTEN전염조PTEN단백급PTEN mRNA표체교정상조급공재체전염조세포균현저상승(균P<0.05).재정상조、TGF-β1자격조、PTEN+TGF-β1조、공재체+TGF-β1조중,TGF-β1자격조급공재체+TGF-β1조교정상조E-cadherin표체명현하강(균P<0.05),이a-SMA급p-Akt표체현저상승(균P<0.05);PTEN+TGF-β1조교TGF-β1자격조급공재체+TGF-β1조a-SMA급p-Akt표체명현하강(균P<0.05).이E-cadherin표체상승(균P<0.05);각조세포Akt표체수평차이불명현(P>0.05).결론 PTEN통과조지PI3K/Akt통로활화이억제TGF-β1유도적신소관상피세포전분화.
Objective To investigate the inhibitory effects of overexpression of PTEN on renal epithelial-mesenchymal trans-differentiation induced by TGF-β1,and the signaling transduction mechanism.Methods HKC cells were transfected with GFP-PTEN via lipofectAMINE2000.The efficiency of transfection was detected by fluorescence microscope.The expression of PTEN protein and mRNA in the translected cells was detected by Western blot and RT-PCR respectively.The experiment was divided into four groups:normal group,TGF-β1 stimulation group,GFP-PTEN+TGF-β1 group and empty vector+TGF-β1 group.The expression of E-cadherin,a-SMA,Akt and p-Akt was detected by Western blot.Results Most ceils transfected with GFP-PTEN expressed GFP.The expression of PTEN protein and mRNA was strongly increased when HKC cells were transfected with GFP-PTEN(all P<0.05).In both TGF-β1 stimulation group and empty vector+TGF-β1 group,the expression level of E-cadherin was lower(all P<0.05),while that of p-Akt and a-SMA was higher than in normal group(both P<0.05).The expression level of p-Akt and a-SMA in GFP-PTEN+TGF-β1 group was Iower(both P<0.05),while that of E-cadherin was higher than in TGF-β1 stimulation group and empty vector+TGF-β1 group(both P<0.05).The expression of Akt was similar in the four groups.Conclusion Overexpression of PTEN can inhibit renal epithelial-mesenehymal trans-differentiation induced by TGF-β1 through suppressing the activation of PI3K/Akt signal pathway.
