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• 基础医学 • 上一篇    下一篇

siRNA抑制人前列腺癌PC-3细胞株VEGF的表达及细胞增殖

李然伟1,王 珂2,杨泽成3,刘永红4,吕佳音5,吴丹华*,赵燕颖6   

  1. (1.吉林大学第二医院泌尿外科,吉林 长春 130041;2.吉林大学第二医院呼吸内科,吉林 长春 130041;3. 吉林大学中日联谊医院普外科,吉林 长春 130033;4.吉林省肿瘤医院,吉林 长春 130012;5.吉林大学中日联谊医院
  • 收稿日期:2007-09-05 修回日期:1900-01-01 出版日期:2008-07-28 发布日期:2008-07-28
  • 通讯作者: 赵燕颖

Inhibition of siRNA on VEGF expression in PC-3 cell line and cell proliferation

LI Ran-wei1,WANG Ke2,YANG Ze-cheng3,LIU Yong-hong4,LU Jia-yin5,WU Dan-hua*,ZHAO Yan-ying6   

  1. (1.Department of Urinary surgery,Second Hospital,Jilin University,Changchun 130041,China;2.Department of Respiratory Disease,Second Hospital,Jilin University,Changchun 130041,China;3. Department of General Surgery,China-Japan Union Hospital,Jilin University,Changchun 130033,China;4. Tumor Hospital of Jilin Province,Changchun 130012,China;5. Department of Orthopedics,China-Japan Union Hospital,Jilin University,Changchun 130033,China;6. Department of Electrodiagnosis,Second Hospital,Jilin University,Changchun 130041,China)
  • Received:2007-09-05 Revised:1900-01-01 Online:2008-07-28 Published:2008-07-28
  • Contact: ZHAO Yan-ying

摘要: 目的:研究小干扰RNA(siRNA)对PC-3人前列腺癌细胞血管内皮生长因子(VEGF)基因表达的抑制作用及抑制癌细胞增殖和诱导凋亡的作用。方法:体外合成特异性靶向人VEGF基因的siRNA,并转染到PC-3细胞中;RT-PCR和Western blotting检测siVEGF对VEGF基因和蛋白表达的抑制作用;MTT法检测siVEGF对癌细胞增殖抑制作用,流式细胞术检测siVEGF诱导细胞凋亡的作用。结果:转染siVEGF1和siVEGF2组VEGF mRNA 表达水平与空白对照组比较分别下调了60.32%和70.73%;VEGF蛋白表达水平与空白对照组比较明显受到抑制,最高抑制率达72%;转染阴性对照质粒组VEGF基因和蛋白表达水平都无明显改变。转染siVEGF1和siVEGF2的细胞增殖受到抑制,增殖抑制率分别为49.5%和55.3 %,细胞的凋亡率分别为33.9%和42.0%。结论:构建的siRNA VEGF能特异有效下调VEGF基因的表达,瞬时转染siRNA VEGF能有效抑制人前列腺癌细胞PC-3增殖并诱导其凋亡。

关键词: 血管内皮生长因子, 前列腺肿瘤, 逆转录聚合酶链反应, 流式细胞术

Abstract: To study the inhibitory effect of small interfering RNA(siRNA) on VEGF expression in prostate cancer cell line PC-3 and to observe its influences in cell proliferation and apoptosis. Methods Two siRNAs against the VEGF gene were designed and transfected into PC-3 cells respectively,VEGF gene and protein expressions were measured by RT-PCR and Western blotting.Cell proliferation was measured with MTT assay, apoptosis was measured with flow cytometry(FCM). Results The VEGF mRNA expressions were down-regulated by 60.32 % and 70.73%,and the VEGF protein expressions were maximally down-regulated by 72% after transfecting with siVEGF1 and siVEGF2, respectively. The inhibitory rates of cell proliferation were 49.5% and 55.3 %;the apoptotic rates after transfecting were 33.9 % and 42.0%. Conclusion siRNA against VEGF could effectively down-regulate VEGF expression in PC-3 cell line,it can inhibit cell growth and induce apoptosis.

Key words: vascular endothelial growth factor, prostate neoplasms, flow cytometry, reverse transcriptase polymerase chain reaction

中图分类号: 

  • R737.25