国家自然科学基金资助项目(30000083),上海市科委项目(00JC 14042)和教育部分子医学重点实验室资助项目.
This work was supported by grants from The National Natural Sciences Foundation of China (30000083) , Science and Technology Bureau of Shanghai Municipal Government (00JC 14042) and The Key Laboratory of Molecular Medicine, Ministry of Education, China.
表皮钙粘蛋白(E-cadherin)阴性的乳腺癌细胞株MDA-MB-231和MDA-MB-435转染野生型表皮钙粘蛋白基因,通过流式细胞仪测量细胞周期发现表皮钙粘蛋白阳性细胞生长变慢,更多细胞停滞在G0/G1期,蛋白质印迹证实由G0/G1期进入S期的重要调控分子细胞周期蛋白-D1(cyclin D1)下降了,并发现表皮钙粘蛋白还能降低直接激活细胞周期蛋白-D1基因转录的β-连环蛋白的蛋白质浓度.蛋白激酶B(PKB)能通过抑制糖原合成激酶-3β(GSK-3β)的活性来抑制β-连环蛋白降解,并在乳腺癌高转移细胞株中普遍过表达,其表达同样受到了表皮钙粘蛋白的抑制.并且在表皮钙粘蛋白阳性细胞中,作为PKB上游信号分子并能激活PKB的粘着斑激酶 (FAK) 和整联蛋白相关激酶(ILK)蛋白量也发生下降,能抑制PKB激活的PTEN蛋白量却增加了.结果显示,表皮钙粘蛋白能通过降低乳腺癌细胞中的PKB蛋白浓度,并通过上游信号分子抑制PKB的激活,进而降低PKB对β-连环蛋白降解的抑制作用,导致β-连环蛋白直接调控的靶基因细胞周期蛋白D1的表达量下降,引起更多的细胞停止在G0/G1期.
E-cadherin-negative human breast carcinoma cell lines, MDA-MB-231 and MDA-MB-435 were transfected with wild-type E-cadherin cDNA. Flow cytometry showed that E-cadherin-positive transfectants grew slower than the control cells and more cells were relayed in G0/G1 phase. Western blot showed that it was due to down-regulation of protein concentration of cyclin D1 and β-catenin, the cyclin D1 gene transcriptional regulator. At the meantime, PKB protein level, which can inhibit β-catenin destruction through GSK-3β, was also down-regulated. As the PKB activators, FAK and ILK protein levels were decreased and PKB inhibitor, PTEN was increased by positive expression of E-cadherin. Therefore, E-cadherin can inhibit PKB activity by down-regulation of FAK, ILK and up-regulation of PTEN. As a result, β-catenin and cyclin D1 protein level increased and more cells were retarded in G0/G1 phase.
吴衡,沈敏雄,梁玉龙,段玲玲,王丽影,徐贞,查锡良.阳性表达表皮钙粘蛋白增加G0/G1期人乳腺癌细胞比例及其分子机制[J].生物化学与生物物理进展,2003,30(3):435-441
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