PSMA对LNCaP细胞生长、迁移及ERK蛋白活性的影响  

Effect of prostate-specific membrane antigen on phospho-ERK,growth and migration of prostate cancer LNCaP cells

作　　者：郭正辉[1] 黄海[1] 杜涛[2] 许可慰[1] 曹亿[1] 陈杰青[1] 董文[1] 姚友生[1] 林天歆[1] 谢文练[1] 江春[1] 韩金利[1] 黄健[1] 

机构地区：[1]中山大学孙逸仙纪念医院泌尿外科,广东广州510120 [2]中山大学孙逸仙纪念医院产科实验室,广东广州510120

出　　处：《中国病理生理杂志》2011年第6期1090-1096,共7页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.05300720No.10151008901000070)

摘　　要：目的:利用我们已经成功沉默前列腺特异性膜抗原(PSMA)的LNCaP前列腺癌细胞株,探讨PS-MA对LNCaP细胞磷酸化胞外信号调节激酶(ERK)及细胞生长、迁移的影响,为进一步研究PSMA在前列腺癌发展中的作用提供理论基础。方法:实验对象包括携带可稳定抑制PSMA表达siRNA慢病毒的LNCaP细胞组(实验组),携带对任何基因无干扰作用siRNA慢病毒的LNCaP细胞组(空转组),同时建立未进行处理的普通LNCaP细胞组(对照组),分别对在一般培养基及添加ERK蛋白上游抑制剂的3组细胞,使用Western blotting和细胞免疫化学的方法检测MAPK/ERK蛋白活性,并用MTT描绘细胞生长曲线、Transwell观察细胞迁移情况。结果:在一般培养基的3组细胞中,Western blotting提示实验组磷酸化ERK蛋白表达明显低于对照组和空转组;免疫细胞化学结果显示实验组染色明显比对照组和空转组弱,阳性细胞数较少;MTT绘制生长曲线,得到实验组细胞的增殖生长能力较对照组、空转组降低;Transwell结果提示实验组较对照组和空转组细胞的增殖迁移能力降低。在ERK磷酸化被抑制的情况下,3组细胞磷酸化ERK蛋白均低表达,MTT及Transwell检测显示其生长迁移能力都处于低水平,且与在一般培养基中实验组的效应类似。结论:初步发现PSMA可能通过上调前列腺癌LNCaP细胞ERK蛋白的活性,从而在其生长、迁移中起正向调节作用。AIM：To study the expression of prostate-specific membrane antigen（PSMA） on the level of phospho-ERK,cell growth and migration of prostate cancer LNCaP cells.METHODS： The method of silencing PSMA was established by lentivirus-mediated RNAi in our early experiment.The cells were divided into 3 groups.In experimental group,the expression of PSMA in LNCaP cells was stably blocked by lentivirus-mediated RNAi.In negative control group,the cells were transfected with lentivirus-mediated control RNAi（without any interference to PSMA）.The normal LNCaP cells served as blank control.The cells in these 3 groups were cultured in both 2 environments： normal medium and medium with PD98059（an inhibitor of ERK phosphorylation）.The phospho-ERK was detected by Western blotting and immunocytochemistry.Furthermore,the growth and migration of the cells were evaluated by MTT and transwell assays,respectively.RESULTS： In normal medium,the expression of phospho-ERK was attenuated in experimental group（P0.05） and the quantity of ＂positive＂ cells was less than those in other 2 groups（P0.05）.Furthermore,the growth curves of the cells showed that the growth ability in experimental group was significantly decreased（P0.05,after 48 h） and the migration ability in experimental group was reduced（P0.05）.In the inhibitory medium,the cells in all 3 groups expressed phospho-ERK at a lower level.Moreover,the abilities of growth and migration in these 3 groups were poorly displayed.These inhibitory effects on phosphorylation of ERK were similar to the cells in experimental group cultured in normal medium.CONCLUSION： PSMA may play a role in up-regulation of phospho-ERK and it may take an advantage in growth and migration of prostate cancer LNCaP cells.

关 键 词：前列腺肿瘤 前列腺特异性膜抗原 有丝分裂素激活蛋白激酶类 

分 类 号：R737.25[医药卫生—肿瘤]