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),G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。
Objective： To observe the effects of pAAV-siE2F3 mediated E2F3 gene on silencing prostate cancer cell line LN- CAP. Methods： pAAV-siE2F3 plasmids were transfected into prostate cancer cells LNCAP. After LNCAP cells were transfected, the in- terference 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 LN- CAP cells were transfected with pAAV-siE2F3 plasmid, the apoptosis index of LNCAP cells increased obviously （ P 〈 0.01 ）, the num- ber of cells during G~ phase increased （ P 〈 0.01 ）, and the number of cells during S phase decreased evidently （ P 〈 0.01 ）. The expres- sion 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.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2012年第2期70-73,共4页
Chinese Journal of Clinical Oncology
基金
天津市科技计划项目(编号:09JCYBJC27800)资助
