雄激素非依赖性前列腺癌细胞雄激素受体靶向降解的实验研究  

Targeted Degradation of Androgen Receptors in Androgen-Independent Prostate Cancer Cells:An Experimental Study

作　　者：施云峰[1,2] 唐悦清[1,2] 孙晓文[1,2] 于殿军[1,2] 韩邦旻[1,2] 洪艳[1,2] 夏术阶[1,2] 

机构地区：[1]上海交通大学附属第一人民医院泌尿外科 [2]上海交通大学泌尿外科研究所,上海200080

出　　处：《中华男科学杂志》2009年第12期1059-1063,共5页National Journal of Andrology

基　　金：国家自然科学基金资助项目(30600618);上海市科委项目(07QA14037)~~

摘　　要：目的:研究合成的嵌合分子DHT-PROTAC通过泛素蛋白酶体途径对雄激素非依赖性前列腺癌C4-2B细胞表达雄激素受体(AR)的靶向降解作用,以及AR降解后该细胞分泌、增殖和凋亡的变化。方法:实验细胞分3组:空白对照组(RPMI1640培养液)、溶剂对照组[0.1%二甲基亚砜(DMSO)]及实验组(100μmol/LDHT-PRO-TAC);制备细胞爬片,免疫组化和Western印迹检测C4-2B细胞中AR蛋白和Caspase-3表达情况的改变;ELISA法测定细胞培养液上清中PSA的浓度改变。噻唑兰(MTT)比色法检测细胞增殖抑制,细胞计数并绘制生长曲线观察细胞增殖能力。结果:与空白对照组和溶剂对照组相比,DHT-PROTAC处理后,C4-2B细胞中表达的AR蛋白明显减少,Western印迹检测结果表现为信号减弱(P<0.05);免疫组化见AR阳性细胞明显减少,阳性率降低达40%;与空白对照组和溶剂对照组相比,DHT-PROTAC处理后细胞培养上清液中PSA浓度降低约60%(P<0.05)。MTT试验显示随着DHT-PROTAC处理时间延长,对C4-2B细胞的抑制率逐渐增大,细胞生长呈时间依赖性抑制(P<0.05)。结论:嵌合分子DHT-PROTAC能靶向降解C4-2B细胞表达的AR,抑制细胞分泌PSA,并抑制该细胞的体外生长和增殖。Objective： To investigate targeted degradation of the androgen receptor （AR） by chimeric molecules （DHT-PROTAC） via the ubiquitin-proteasome pathway in androgen-independent prostate cancer CA-2B cells, and explore the proliferation, secretion and apoptosis of the treated cells. Methods： CA-2B cells were treated with DHT-PROTAC, and then the expressions of the AR protein and caspase3 in the CA-2B cells were detected by immunohistochemistry and Western blot. The concentration of PSA in the supematant was examined by ELISA. The ceils were counted and their proliferation analyzed by a growth curve. The inhibitory effect on the growth of CA-2B ceils was evaluated by MTT assay. Results： Compared with the control group, the DHT-PROTAC-treated group showed an obviously decreased expression of AR proteins with a significant attenuation of the band signals （ P 〈 0.05 ）, a 40% reduction of the AR-positivc cells and a 60% decrease of the PSA concentration in the supematant （P 〈 0.05 ）. DHT-PROTAC exhibited an inhibitory effect on the CA-2B cells in a time-dependant manner （P 〈 O. 05 ）. Conclusion： The chimeric molecule （DHT-PROTAC） can target the degradation of androgen receptors, reduce the secretion of PSA and repress the in vitro growth of CA-2B cells.

关 键 词：雄激素受体 雄激素非依赖性前列腺癌 蛋白水解作用 嵌合分子 

分 类 号：R737.25[医药卫生—肿瘤]