国际肿瘤学杂志

国际肿瘤学杂志 ›› 2015, Vol. 42 ›› Issue (1): 14-17.doi: 10.3760/cma.j.issn.1673-422X.2015.01.004

• 论著 • 上一篇    下一篇

shRNA靶向干扰NHERF1对PC-3M前列腺癌细胞生物学行为的影响

马强, 郑君芳, 焦延娜, 高红林, 李德冠, 刘鉴峰, 刘强, 宋娜玲   

  1. 300192天津，北京协和医学院 中国医学科学院放射医学研究所 天津市放射医学与分子核医学重点实验室；首都医科大学生物化学与分子生物学系（郑君芳）；国家人口计生委科学技术研究所遗传中心（焦延娜）
  • 收稿日期:2014-07-09 修回日期:2014-10-18 出版日期:2015-01-08 发布日期:2015-01-07
  • 通讯作者: 宋娜玲 E-mail:nalingsong@sina.com
  • 基金资助:

    中国医学科学院放射医学研究所探索基金（ST1432）

RNA interference gene therapy targeting NHERF1 inhibits proliferation of prostate cancer cell line PC-3M

Ma Qiang, Zheng Junfang, Jiao Yanna, Gao Honglin, Li Deguan, Liu Jianfeng, Liu Qiang, Song Naling   

  1. Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Peking Union Medical College & Chinese Academy of Medical Science, Tianjin 300192, China
  • Received:2014-07-09 Revised:2014-10-18 Online:2015-01-08 Published:2015-01-07
  • Contact: Song Naling E-mail:nalingsong@sina.com

PDF

1344

摘要： 目的 探讨shRNA靶向干扰钠氢交换调节因子（NHERF）1后对前列腺癌细胞PC-3M生物学行为的影响。方法 将pSuper.puro NHERF1 shRNA质粒和阴性对照质粒pSuper.puro luciferase shRNA分别转染人PC-3M前列腺癌细胞，经嘌呤霉素筛选，建立稳定低表达NHERF1的PC3M细胞株。经Western印迹验证后，采用四甲基偶氮唑蓝（MTT）法检测肿瘤细胞增殖活性，采用划痕实验检测NHERF1-shRNA对前列腺癌PC-3M细胞迁移能力的影响，采用流式细胞仪检测NHERF1shRNA对PC3M细胞凋亡的影响，采用Western印迹检测NHERF1shRNA对凋亡相关蛋白Bax及Bcl2表达的影响。结果 转染NHERF1shRNA组比未转染及空载组细胞的增殖能力明显降低（F=62.63，P=0.004）；转染NHERF1shRNA组比未转染及空载组细胞的迁移率明显降低。相比于未转染PC-3M细胞和转染阴性对照质粒细胞，转染NHERF1shRNA组细胞的凋亡百分比明显增加［(2.55±0.92)%和（11.98±3.28）%］，达4倍以上(t=-2.392，P=0.001)；Bcl2蛋白的表达降低，Bax蛋白的表达升高。结论 靶向NHERF1的序列特异性shRNA可抑制PC-3M细胞的增殖和迁移，并促进细胞凋亡。

关键词: 前列腺肿瘤, 细胞凋亡, NHERF1

Abstract: Objective To investigate the effect of NHERF1shRNA on malignant behaviors of prostate cancer cell PC3M. MethodspSuper.puro NHERF1 shRNA vector and negative control (pSuper.puro luciferase shRNA) were transfected into prostate cancer cell PC3M. Then the cell line stably knockdown NHERF1 was obtained and verified by Western blot analysis. Methyl thiazolyl thiazolium (MTT) was used to detect the proliferative activity of PC3M cell. The cell migration abilities were examined by wound healing assay. Flow cytometry method was applied to detect the effect of NHERF1shRNA on apoptosis of PC3M cell line. The expressions of Bax and Bcl2 in NHERF1shRNA cells were detected by Western blot. ResultsCompared with PC3M cells and the cells which were integrated of empty vectors, NHERF1shRNA could weaken the proliferation of PC3M cells significantly (F=62.63, P=0.004), and significantly repress PC3M cells migration, and increase the apoptosis percent of PC3M cell line apparently ［(2.55±0.92)% and (11.98±3.28)%］, up to 4 times (t=-2.392, P=0.001). NHERF1shRNA could reduce the expression of Bcl2 and increase the expression of Bax. ConclusionNHERF1shRNA can inhibit the proliferation and migration of PC3M cell, and promote its apoptosis.

Key words: Prostatic neoplasms, Apoptosis, NHERF1