CAJ | 학술논문

为了研究蛋白激酶B(PKB)对上皮钙粘着蛋白(E-cadherin)的调节,使用了用胰岛素处理的野生型SMMC 7721细胞及稳定表达持续激活PKB的SMMC 7721细胞株(Gag-PKB/SMMC 7721).用RNA印迹法和蛋白质印迹法检测细胞E-cadherin 表达,发现通过胰岛素刺激或在细胞中表达持续激活PKB从而增加PKB活性,不影响E-cadherin的转录和蛋白质合成,但用流式细胞术和免疫荧光定位E-cadherin,则发现PKB活性增加能明显驱动E-cadherin到细胞表面,从而导致部分通过E-cadherin途径的细胞粘聚增加和细胞调亡的抑制.因此,我们提供新的证据表明,增加PKB活性可驱动有功能的E-cadherin分子到细胞表面.
위료연구단백격매B(PKB)대상피개점착단백(E-cadherin)적조절,사용료용이도소처리적야생형SMMC 7721세포급은정표체지속격활PKB적SMMC 7721세포주(Gag-PKB/SMMC 7721).용RNA인적법화단백질인적법검측세포E-cadherin 표체,발현통과이도소자격혹재세포중표체지속격활PKB종이증가PKB활성,불영향E-cadherin적전록화단백질합성,단용류식세포술화면역형광정위E-cadherin,칙발현PKB활성증가능명현구동E-cadherin도세포표면,종이도치부분통과E-cadherin도경적세포점취증가화세포조망적억제.인차,아문제공신적증거표명,증가PKB활성가구동유공능적E-cadherin분자도세포표면.
In order to study the regulation of E-cadherin by protein kinase B (PKB), wild type SMMC 7721 hepato-carcinoma cells and a Gag-PKB/SMMC 7721 cell line where PKB activity is markedly increased compared with control cells were used. Interestingly, increasing PKB activity via insulin stimulation or Gag-PKB transfection does not enhance the E-cadherin in the level of mRNA and that of protein by using Northern blot and Western blot analysis, but markedly drives E-cadherin protein to cell surface by using flow cytometry analysis and immunofluorescence analysis localization of E-cadherin, which resulted in the increase of cell aggregation and the inhibition of cell apoptosis mostly via E-cadherin. Therefore, new evidences that elevation of PKB activity could drive functional E-cadherin molecule to cell surface are provided.