Nkx3.1和p27协同诱导激素抵抗性前列腺癌PC3细胞株凋亡的研究  

Synergistic effect of Nkx3.1 and p27 Proteins in Human prostate cancer cells

作　　者：王平[1] 陈昭典[1] 

机构地区：[1]浙江大学医学院附属第一医院泌尿外科,浙江杭州310003

出　　处：《健康研究》2011年第4期244-249,共6页Health Research

基　　金：浙江省医药卫生科学研究基金资助(2008A46)

摘　　要：目的探讨人类同源框基因Nkx3.1和细胞周期素依赖激酶抑制剂p27诱导激素抵抗前列腺癌PC3细胞凋亡的协同作用。方法构建Nkx3.1和p27的真核表达质粒载体,恢复表达PC3细胞Nkx3.1或/和p27表达。Nkx3.1或p27单独和二者结合转染PC3细胞后,检测细胞增殖、细胞周期和细胞凋亡,同时分析了凋亡相关蛋白表达变化。结果结合Nkx3.1和p27显示协同抗增殖效应。与对照组相比,转染Nkx3.1的PC3细胞株增殖无显著性差异,而转染p27的PC3细胞株增殖在48 h时间点存在显著性差异(P<0.05),二者共转染PC3细胞增殖在24 h(P<0.05)和48 h(P<0.01)时间点均存在显著性差异。p27使阻滞在G0/G1期PC3细胞数增加,当共转染Nkx3.1后,这种细胞周期阻滞效应显著增强。Bcl-2、Bax、caspase-3、PARP等蛋白表达和流式细胞检测发现共转染组引起PC3细胞凋亡亦存在协同效应,协同效应主要表现为caspase-3前体被激活、Bcl-2表达下降、Bax表达上升,PARP劈裂片断表达上升。结论结合Nkx3.1和p27可协同抑制PC3细胞增殖、阻滞细胞在G0/G1期、促进凋亡。凋亡相关蛋白分析显示Bcl-2表达抑制、Bax表达增强、caspase-3激活和PARP劈裂片段表达增加。研究显示二者结合在基因治疗激素抵抗前列腺癌有潜在的应用价值。Objective To investigate the synergistic effect of the homeobox gene Nkx3.1 and the cyclin-dependent kinase inhibitor p27 in PC3 human prostate cancer cells. Methods Two recombinant plasmids （ pcDNA3. 1-Nkx3. 1-His and pcDNA3. 1-p27-HA） were transfected to PC3 cells. The analysis of cell proliferation, cell cycle and apoptosis were performed to test the synergistic effect of Nkx3.1 and p27 proteins. Results Overexpression of p27 protein, not Nkx3.1, promoted PC3 cells＇ proliferation. Combination of Nkx3. 1 and p27 had a synergistic anti-proliferative effect at 24 （P 〈 0.05） and 48 （P 〈 0.01 ） hours, p27 protein ceased cells to G0/G1 phase and addition of Nkx3.1 increased this action. FACS data showed that pro-caspase-3 and Bcl-2 proteins decreased and Bax protein increased in PC3 cells when overexpressing of p27 and Nkx3. 1 proteins. Conclusion The combination of Nkx3. 1 and p27 displayed synergistic multiple antieancer effects in vitro, with the mechanism of anti-proliferation and stimulating apoptosis. This combination may be a potential gene therapy method for androgen-independent prostate cancer.

关 键 词：NKX3.1 P27 前列腺癌细胞 增殖 凋亡 

分 类 号：R737.25[医药卫生—肿瘤]