陈涛, 胡海龙, 孙岩, 吴长利. 以pAAV为载体的RANi沉默E2F3对前列腺癌LNCAP细胞系的影响[J]. 中国肿瘤临床, 2012, 39(2): 70-73. DOI: 10.3969/j.issn.1000-8179.2012.02.003
引用本文: 陈涛, 胡海龙, 孙岩, 吴长利. 以pAAV为载体的RANi沉默E2F3对前列腺癌LNCAP细胞系的影响[J]. 中国肿瘤临床, 2012, 39(2): 70-73. DOI: 10.3969/j.issn.1000-8179.2012.02.003
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Tao CHEN, Hailong HU, Yan SUN, Zhangli WU. Effects of Silencing E2F3 Expression Using RNAi with pAAV Virus Exprsessing Vector in Prostate Cancer CeU Line LNCAP[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2012, 39(2): 70-73. DOI: 10.3969/j.issn.1000-8179.2012.02.003
Citation: Tao CHEN, Hailong HU, Yan SUN, Zhangli WU. Effects of Silencing E2F3 Expression Using RNAi with pAAV Virus Exprsessing Vector in Prostate Cancer CeU Line LNCAP[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2012, 39(2): 70-73. DOI: 10.3969/j.issn.1000-8179.2012.02.003
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以pAAV为载体的RANi沉默E2F3对前列腺癌LNCAP细胞系的影响

Effects of Silencing E2F3 Expression Using RNAi with pAAV Virus Exprsessing Vector in Prostate Cancer CeU Line LNCAP

    摘要
  • 摘要:
      目的  探讨以pAAV为载体的RANi沉默E2F3对前列腺癌LNCAP细胞系的影响及其作用机制。
      方法  用pAAV-siE2F3穿梭质粒转染LNCAP细胞, 流式细胞计数检测转染后细胞凋亡, 细胞周期变化; Westernblot检测转染后E2F3蛋白与Bcl-2蛋白表达的变化, 分析E2F3蛋白与Bcl-2蛋白变化相关性。
      结果  与对照组相比, pAAV-siE2F3穿梭质粒有效的沉默了E2F3蛋白表达(P < 0.01), pAAV-siE2F3穿梭质粒组细胞凋亡明显增加(P < 0.01), G1期细胞明显增加(P < 0.01), S期明显减少(P < 0.01), pAAV-siE2F3穿梭质粒组与对照组相比, Bcl-2蛋白表达明显减少(P < 0.01)。
      结论  本实验初步明确了E2F3在前列腺癌细胞周期调控细胞凋亡方面起的作用, E2F3可能通过影响Bcl-2表达调控细胞凋亡; 为进一步阐明E2F3基因与前列腺癌的关系, 及以E2F3为靶点的前列腺癌基因治疗提供理论基础。

     

    Abstract:
      Objective  To observe the effects of pAAV-siE2F3 mediated E2F3 gene on silencing prostate cancer cell line LNCAP.
      Methods  pAAV-siE2F3 plasmids were transfected into prostate cancer cells LNCAP. After LNCAP cells were transfected, the interference effects were detected by Westen blot. The appoptosis index and cell cycle of LNCAP cells were detected by flow cytometry.
      Results  The results of Western blot indicated that pAAV-siE2F3 plasmid could knock down the expression of E2F3 gene. After LNCAP cells were transfected with pAAV-siE2F3 plasmid, the apoptosis index of LNCAP cells increased obviously (P < 0.01), the number of cells during G1 phase increased (P < 0.01), and the number of cells during S phase decreased evidently (P < 0.01). The expression of Bcl-2 gene decreased apparently (P < 0.01).
      Conclusions  E2F3 plays an important role in the progress of prostate cancer cell cycle. E2F3 regulates cell appotosis index through affectting the expression of Bcl-2 genes, indicating that E2F3 maybe a new target of gene therapy for prostate cancer.

     

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