Yang Wuxiao, Li Pengcheng, Lin Jingyang, Zuo Houjuan, Zuo Ping, Zou Yuanlin, Liu Zhengxiang
Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Department of Cardiology, Zhejiang Provincial People's Hospital, Hangzhou, 310014, China.
J Huazhong Univ Sci Technolog Med Sci. 2012 Jun;32(3):383-388. doi: 10.1007/s11596-012-0066-y. Epub 2012 Jun 9.
Over-expression of CD151 was found to be associated with metastasis and poor prognosis of prostatic carcinoma. This study was designed to examine the mechanism by which CD151 promotes the proliferation and migration of prostatic cancer cells. The pAAV-CD151, pAAV-GFP and pAAV-CD151-AAA mutant plasmids were constructed and used to transiently transfect PC3 cells (a prostatic carcinoma 3 cell line) by the mediation of Fugene HD. Then, the cells were assigned to control group, pAAV-GFP group, pAAV-CD151 group, and pAAV-CD151-AAA group respectively. Cell proliferation was evaluated by using the 3-[4,5-dimet-hylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method. Cell migration assay was performed by using Boyden chambers. The formation of CD151-integrin α3/α6 complex was determined by the method of co-immunoprecipitation. The protein expression levels of CD151 and extracellular signal-regulated kinase (ERK) were measured by Western blotting. The results showed that transfection of pAAV-CD151 or pAAV-CD151-AAA mutant increased the expression of CD151 protein in PC3 cells. Co-immunoprecipitation showed that more CD151-integrin α3/α6 complex was formed in the pAAV-CD151 group than in the control group, the pAAV-GFP group and the pAAV-CD151-AAA mutant group. Furthermore, the proliferative and migrating capacity of PC3 cells was substantially increased in the pAAV-CD151 group but inhibited in the pAAV-CD151-AAA mutant group. CD151 transfection increased the expression of phospho-ERK. Taken together, it was concluded that CD151 promotes the proliferation and migration of PC3 cells through the formation of CD151-integrin complex and the activation of phosphorylated ERK.
研究发现CD151的过表达与前列腺癌的转移及不良预后相关。本研究旨在探讨CD151促进前列腺癌细胞增殖和迁移的机制。构建了pAAV - CD151、pAAV - GFP和pAAV - CD151 - AAA突变体质粒,并通过Fugene HD介导瞬时转染PC3细胞(一种前列腺癌3细胞系)。然后,将细胞分别分为对照组、pAAV - GFP组、pAAV - CD151组和pAAV - CD151 - AAA组。采用3 - [4,5 - 二甲基噻唑 - 2 - 基] - 2,5 - 二苯基四氮唑溴盐(MTT)法评估细胞增殖。使用Boyden小室进行细胞迁移实验。通过免疫共沉淀法测定CD151 - 整合素α3/α6复合物的形成。采用蛋白质印迹法检测CD151和细胞外信号调节激酶(ERK)的蛋白表达水平。结果显示,转染pAAV - CD151或pAAV - CD151 - AAA突变体均可增加PC3细胞中CD151蛋白的表达。免疫共沉淀显示,与对照组、pAAV - GFP组和pAAV - CD151 - AAA突变体组相比,pAAV - CD151组形成了更多的CD151 - 整合素α3/α6复合物。此外,pAAV - CD151组中PC3细胞的增殖和迁移能力显著增强,而pAAV - CD151 - AAA突变体组中则受到抑制。CD151转染增加了磷酸化ERK的表达。综上所述,得出结论:CD151通过形成CD151 - 整合素复合物和激活磷酸化ERK来促进PC3细胞的增殖和迁移。