摘要
目的:探讨多西紫杉醇治疗男性雄激素抵抗前列腺癌(castration-resistant prostate cancer,CRPC)的分子机制。方法:前列腺癌细胞株LNCaP、PC-3和CW22-rv1体外培养后,通过蛋白质印迹、细胞转染、荧光素酶分析、细胞存活率分析等试验分析多西紫杉醇(Docetaxel,Doc)处理后细胞株的存活、AR及p-c-jun的表达情况及其与细胞生存的关系,同时实时定量PCR检测相应mRNA的表达情况。结果:多西紫杉醇对不同前列腺癌细胞株的敏感性不同,其中PC-3细胞最敏感,CW22-rv1和LNCaP细胞中度敏感,其敏感性与p-c-jun表达呈负相关。转染c-jun基因可降低细胞对多烯紫杉醇的敏感性,而PC-3细胞转染c-jun和AR基因后则可以使细胞恢复到中等程度敏感性,细胞存活率为30%。长期暴露于Bicalutmide(比卡鲁胺)后的LNCaP细胞经Doc处理后PSA蛋白表达增加,AR蛋白表达水平降低,AR的mRNA却增加。结论:p-c-jun降低前列腺癌细胞株对多西紫杉醇的敏感性,而AR可以增加前列腺癌细胞株对多西紫杉醇的反应,AR的上调降低c-jun/p-c-jun的转录活性,从而增加前列腺癌细胞株对Doc的反应,可能是Doc治疗前列腺癌的机制之一。
Objective： The present study investigated the biochemical mechanism of docetaxel （Doc） for the treatment of castra- tion-resistant prostate cancer （CRPC）. Methods： The prostate cancer cell lines PC-3, LNCaP, and CW22-rvl were grown in vitro. The expression of c-Jun and the androgen receptor （AR）, the relationship between their expression levels, and the viability of LNCaP, PC-3, and CW22-rvl cells after exposure to Doc were studied in vitro by immunoblot, luciferase, and viability assays. Furthermore, the ex- pression of AR mRNA was analyzed by quantitative real-time polymerase chain reaction （RT-PCR）. Results： Doc therapy elicited dif- ferent responses in the different cell lines. PC-3 cells were the most sensitive to treatment, whereas a moderate response was observed in CW22-rvl and LNCaP cells. The sensitivity and response to Doc was negatively correlated with the level of phosphorylated c-Jun （p-c-Jun） expression. Transfection of the c-jun gene into cell lines decreased their response to Doc treatment, whereas cotransfection of the c-jun and AR genes restored the sensitivity of PC-3 to a moderate degree, with 30% viability was 30%. After long-term exposure to bicalutamide （cas）, the Doc-treated LNCaP cells demonstrated increased levels of the prostate specific antigen （PSA）. Likewise the AR protein levels decreased with increasing AR mRNA expression. Conclusion： This study demonstrated that the level of p-c-Jun de- creased the response of PC cells to Doc treatment, but the level of AR increases PC cells response to Doc treatment. The increased AR level inhibits c-Jun/p-c-Jun transcriptional, which decreases the protective effects of Doc in PC cells.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2013年第8期444-448,共5页
Chinese Journal of Clinical Oncology
基金
云南省卫生科技计划项目(编号:2010NS053)资助~~
