Wang Ping, Liu Ben, Luo Jin-Dan, Zhang Zhi-Gen, Ma Qi, Chen Zhao-Dian
Department of Urology, The First Affiliated Hospital, Medical College of Zhejiang University, Hangzhou, 310003, China.
Asian J Androl. 2007 May;9(3):353-60. doi: 10.1111/J.1745-7262.2007.00278.X.
To explore whether the anti-tumor action of 17beta-estradiol is enhanced by re-expression of the homeodomain transcription factor Nkx3.1 in PC3 human prostate cancer cells.
PC3 cells were stably transfected with pcDNA3.1-Nkx3.1-His vector, which carries a full-length cDNA of human Nkx3.1. The PC3 cells stably transfected with vector pcDNA3.1 were set as a control. The expression of Nkx3.1 protein in the cells was confirmed by Western blot analysis. The effect of Nkx3.1 on cell proliferation of PC3 cells was examined with MTT assay. The antiproliferative and apoptotic effects of 17beta-estradiol alone or in combination with Nkx3.1 were estimated on PC3 cells by using MTT growth tests and flow cytometric analyses. The expression of apoptosis-related proteins was analyzed using Western blotting.
The plasmid carrying Nkx3.1 gene induced high expression of Nkx3.1 protein in PC3 cells. The re-expression of exogenous Nkx3.1 did not cause a significant reduction in cellular proliferation, whereas the expression of Nkx3.1 enhanced the 17beta-estradiol anti-proliferative effect in PC3 cells. Nkx3.1 expression promoted 17beta-estradiol-induced apoptosis of PC3 cells, as shown by analysis of Bcl-2, Bax, Caspase-3 and poly (ADP-ribose) polymerase expression.
The present study demonstrates that re-expression of Nkx3.1 enhances 17beta-estradiol anti-tumor action in PC3 human prostate cancer cells. The in vitro study suggests that re-expression of Nkx3.1 is worthy of further consideration as an adjuvant treatment of androgen independent prostate cancer with estrogen anti-tumor therapies.
探讨在PC3人前列腺癌细胞中重新表达同源结构域转录因子Nkx3.1是否能增强17β-雌二醇的抗肿瘤作用。
用携带人Nkx3.1全长cDNA的pcDNA3.1-Nkx3.1-His载体稳定转染PC3细胞。将稳定转染载体pcDNA3.1的PC3细胞设为对照。通过蛋白质免疫印迹分析确认细胞中Nkx3.1蛋白的表达。用MTT法检测Nkx3.1对PC3细胞增殖的影响。通过MTT生长试验和流式细胞术分析评估单独或与Nkx3.1联合使用的17β-雌二醇对PC3细胞的抗增殖和凋亡作用。用蛋白质免疫印迹法分析凋亡相关蛋白的表达。
携带Nkx3.1基因的质粒在PC3细胞中诱导Nkx3.1蛋白高表达。外源性Nkx3.1的重新表达未导致细胞增殖显著降低，而Nkx3.1的表达增强了17β-雌二醇对PC3细胞的抗增殖作用。如对Bcl-2、Bax、Caspase-3和聚（ADP-核糖）聚合酶表达的分析所示，Nkx3.1表达促进了17β-雌二醇诱导的PC3细胞凋亡。
本研究表明，Nkx3.1的重新表达增强了17β-雌二醇在PC3人前列腺癌细胞中的抗肿瘤作用。体外研究表明，Nkx3.1的重新表达作为雌激素抗肿瘤疗法辅助治疗雄激素非依赖性前列腺癌值得进一步考虑。
