天津医药

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shRNA 沉默CtBP2 基因对前列腺癌细胞的增殖作用

刘妍1,徐勇2,张志宏3,高超1   

  1. 1. 天津市泌尿外科研究所
    2. 天津医大二院泌尿外科
    3. 天津医科大学第二医院泌尿外科
  • 收稿日期:2014-03-03 修回日期:2014-06-16 出版日期:2014-10-15 发布日期:2014-10-15
  • 通讯作者: 刘妍

Study of Effect of CtBP2 Knockout through shRNA on Proliferation of Prostate Cancer Cells

  • Received:2014-03-03 Revised:2014-06-16 Published:2014-10-15 Online:2014-10-15
  • Contact: LIU Yan

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摘要：

【摘要】目的 探讨RNA 干扰技术沉默羧基末端结合蛋白（CtBP）2 基因表达对前列腺癌PC3 细胞增殖能力的影响。方法 实验分为空白对照组、转染空质粒组、转染shRNA 组。设计并合成针对CtBP2 的3 条特异性短发卡 RNA（shRNA）模板，构建CtBP2-shRNA 重组质粒，转染PC3 细胞。通过RT-PCR 检测CtBP2 mRNA 的表达水平；采用Western blot 法检测CtBP2 蛋白表达的水平，运用MTT 法检测沉默CtBP2 对PC3 细胞体外增殖的抑制作用。结果 将CtBP2-shRNA 转染前列腺癌PC3 细胞后，CtBP2 mRNA 以及蛋白的表达水平均明显下降。沉默CtBP2 表达后，MTT 结果显示转染shRNA 组的PC3 细胞较空白对照组、转染空质粒组细胞增殖能力明显减弱，差异有统计学意义（P < 0.01）。结论 CtBP2-shRNA 可抑制CtBP2 在前列腺癌细胞中的表达并抑制肿瘤细胞生长，提示CtBP2 可作为前列腺癌基因治疗的一个新靶点。

关键词: RNA干扰, 羧基末端结合蛋白, 前列腺肿瘤, 细胞系, 细胞增殖

Abstract:

[Abstract] Objective  To study the effects of shRNA-CtBP2 on the growth of prostate cancer PC3 cells. Methods There were three experimental groups in this study，which include blank control group, empty plasmid transfected group and transfected shRNA group. CtBP2 mRNA sequence is targeted by 3 pairs of designed interfering shRNA to built shRNA-CtBP2 recombinant plasmid then it is transfected into PC3 cells. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot assays were used to detect the transcription and expression levels of CtBP2 mRNA and protein, respectively. PC3l proliferation was measured by MTT assay. Results  Builting shRNA-CtBP2 recombinant plasmid and transfect? ing PC3 cells were successful. Transcription and expression levels of CtBP2 mRNA and protein were significantly decreased in shRNA-CtBP2 transfected PC3 cells. After CtBP2 silencing, cell proliferation was blocked in the shRNA-CtBP2 cells compared to that of blank control group (P<0.01). Conclusion  shRNA-CtBP2 could significantly inhibit CtBP2 expres? sion, suppress the growth of PC3 cells, which suggests that CtBP2 may be a new target for PCa gene therapy.

Key words: RNA interference, C-terminal-binding protein, prostate neoplasms, cell line, cell proliferation