吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (03): 460-463.doi: 10.13481/j.1671-587x.20150305

• 基础研究 • 上一篇    下一篇

CD147 RNA干扰对前列腺癌LNCaP-AI细胞体外侵袭力的影响

方芳1, 张万生2, 唐化勇2, 赵良中1, 陈爽1, 李强1, 王立国2   

  1. 1. 吉林医药学院检验学院医学免疫学教研室, 吉林 吉林 132013;
    2. 吉林医药学院附属医院泌尿外科, 吉林 吉林 132013
  • 收稿日期:2014-09-11 发布日期:2015-08-01
  • 通讯作者: 王立国,主任医师(Tel:0432-64560741,E-mail:urolancet@sina.com) E-mail:urolancet@sina.com
  • 作者简介:方芳(1973-),女,吉林省长春市人,副教授,医学博士,主要从事前列腺癌基础方面的研究。
  • 基金资助:

    国家自然科学基金资助课题(81202031);吉林省科技厅科技发展计划项目资助课题(20130101154JC);吉林省教育厅科研基金资助课题(2013346,2012333,2014360)

Influence of CD147 RNA interference in invasion of prostate cancer LNCaP-AI cells in vitro

FANG Fang1, ZHANG Wansheng2, TANG Huayong2, ZHAO Liangzhong1, CHEN Shuang1, LI Qiang1, WANG Liguo2   

  1. 1. Department of Immunology, Institute of Laboratory Medicine, Jilin Medical College, Jilin 132013, China;
    2. Department of Urinary Surgery, Affiliated Hospital, Jilin Medical College, Jilin 132013, China
  • Received:2014-09-11 Published:2015-08-01

摘要:

目的:探讨CD147对前列腺癌LNCaP-AI细胞体外侵袭力的影响,阐明CD147对LNCaP-AI细胞侵袭作用的机制,为研究前列腺癌的侵袭过程提供依据。方法:利用脂质体2000分别转染靶向CD147基因的shRNA质粒和阴性对照质粒至LNCaP-AI细胞中,经G418筛选获得稳定表达株,分别命名为AI/shCD147(CD147沉默组)和AI/Scramble(阴性对照组)。Western blotting法检测2组细胞中CD147蛋白表达。Transwell实验检测2组LNCaP-AI细胞的体外侵袭力。Western blotting法检测2组细胞中基质金属蛋白酶2 (MMP-2) 蛋白表达水平。结果:Western blotting法,与AI/Scramble组比较,AI/shCD147组LNCaP-AI细胞中CD147蛋白表达水平明显降低。Transwell实验,AI/shCD147组吸光度(A)值(1.43±0.2)明显低于AI/Scramble组(2.08±0.3),差异有统计学意义(P<0.05)。Western blotting法,AI/shCD147组细胞中MMP-2蛋白表达水平(0.46±0.08)明显低于AI/Scramble组(0.85±0.02),差异有统计学意义(P<0.05)。结论:CD147可通过诱导MMP-2分泌促进前列腺癌LNCaP-AI细胞的体外侵袭力,其可能成为前列腺癌基因治疗的新策略。

关键词: 前列腺肿瘤, CD147, RNA干扰, 基质金属蛋白酶, 侵袭

Abstract:

Objective To explore the effect of CD147 on the invasion of prostate cancer LNCaP-AI cells and its possible mechanism in vitro, and to provide basis for research on prostate cancer invasion. Methods The shRNA vector targeting CD147 gene and negative control vector were transfected into LNCaP-AI cells by Lipofectamine 2000, respectively.The stable cell lines were obtained after G418 screening and divided into AI/shCD147 group (experiment group) and AI/Scramble group (control group), respectively.Western blotting method was employed to detect the expression of CD147 protein in the cells in two groups.Transwell invasion assay was used to evaluate the invasion ability of LNCaP-AI cells in vitro.The level of matrix metalloproteinase-2 (MMP-2) was determined by Western blotting method. Results Compared with AI/Scramble group, the CD147 protein in the LNCaP-AI cells in AI/shCD147 group was significantly decreased.The Transwell invasion assay showed that the A value of LNCaP-AI cells in AI/shCD147 group (1.43±0.2) was significantly lower than that in AI/Scramble group (2.08±0.3) (P<0.05).Compared with AI/Scramble group (0.85±0.02), the expression of MMP-2 protein in AI/shCD147 group (0.46±0.08) was significantly decreased (P<0.05). Conclusion CD147 can promote the invasion ability of LNCaP-AI cells in vitro through inducing the MMP-2 secretion.It is demonstrated that targeting CD147 could potentially be a new approach for prostate cancer gene therapy.

Key words: prostate neoplasms, CD147, RNA interference, matrix metalloproteinase, invasion

中图分类号: 

  • R737.25