首页> 中文期刊> 《中国病理生理杂志》 >PSMA对LNCaP细胞生长、迁移及ERK蛋白活性的影响

PSMA对LNCaP细胞生长、迁移及ERK蛋白活性的影响

         

摘要

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 ( P < 0.05 ) and the quantity of "positive" cells was less than those in other 2 groups ( P <0.05 ).Furthermore, the growth curves of the cells showed that the growth ability in experimental group was significantly decreased ( P <0.05, after 48 h ) and the migration ability in experimental group was reduced ( P < 0.05 ).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.%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PSMA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.

著录项

  • 来源
    《中国病理生理杂志》 |2011年第6期|1090-1096|共7页
  • 作者单位

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,产科实验室,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

    中山大学孙逸仙纪念医院,泌尿外科,广东,广州,510120;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 前列腺肿瘤;
  • 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;

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