摘要
恩杂鲁胺耐药(enzalutamide resistance,ENZR)是去势抵抗性前列腺癌(castration-resistant prostate cancer,CRPC)最为常见的问题,患者在使用第二代雄激素受体(androgen receptor,AR)抑制剂恩杂鲁胺(enzalutamide,ENZ)治疗一段时间后,出现对ENZ的抵抗,严重威胁男性健康。但临床尚无治疗前列腺癌ENZ耐药的有效药物。前期基于对恩杂鲁胺的结构优化与改造,并通过细胞活力实验、克隆形成、细胞凋亡检测及蛋白质印迹等分子生物学手段检测发现:LS-13对前列腺癌LNCaP-ENZR和22RV1细胞的半数抑制浓度值分别为17.09和19.90μmol·L^(-1)。随后,通过使用0.1、1、5、10μmol·L^(-1)LS-13处理耐药细胞株12天后,观察结果显示LS-13能够以剂量依赖性方式抑制LNCaP-ENZR和22RV1细胞的体外克隆形成。此外,经过1.25、2.5、5、10μmol·L^(-1)LS-13处理耐药细胞株72 h后,流式细胞术的结果也显示LS-13能够以剂量依赖性方式促进耐药细胞株的凋亡。进一步地,在上述相同浓度条件下,LS-13处理22RV1细胞24 h后,发现其迁移速度显著减缓(P<0.05)。这些结果共同表明,小分子化合物LS-13具有明显抑制ENZR的作用。进一步机制探讨发现,LS-13可通过靶向ARfl/ARv7并促进其蛋白降解,抑制AR对下游靶基因的转录水平,进而抑制ENZR细胞增殖,促进细胞凋亡。该研究为靶向ARfl/ARv7降解,克服ENZ耐药提供候选化合物。
Enzalutamide resistance(ENZR)is the most common issue in castration-resistant prostate cancer(CRPC).After a period of treatment with the second-generation androgen receptor(AR)inhibitor enzalutamide(ENZ),patients develop resistance to ENZ,posing a serious threat to male health.However,there are currently no effective drugs for treating prostate cancer patients with ENZ resistance.Based on the structural optimization and modification of enzalutamide,and through molecular biological methods such as cell viability assays,colony formation,cell apoptosis detection,Western blot and other molecular biology techniques,it was discovered that the half-maximal inhibitory concentration values of LS-13 for prostate cancer LNCaP-ENZR and 22RV1 cells are 17.09 and 19.90μmol·L^(-1),respectively.Subsequently,after treating the resistant cell lines with 0.1,1,5,and 10μmol·L^(-1)of LS-13 for 12 days,the observation results indicated that LS-13 could inhibit the in vitro clonal formation of LNCaP-ENZR and 22RV1 cells in a dose-dependent manner.Furthermore,after treating the resistant cell lines with 1.25,2.5,5,and 10μmol·L^(-1)of LS-13 for 72 h,the results of flow cytometry also showed that LS-13 could promote apoptosis of the resistant cell lines in a dose-dependent manner.Furthermore,under the same concentration conditions described above,after LS-13 treatment of 22RV1 cells for 24 h,it was found that their migration speed was significantly slowed down(P<0.05).These results collectively indicate that the small molecule compound LS-13 has a pronounced effect in inhibiting ENZR.Further mechanism studies found that LS-13 can target ARfl/ARv7 and promote its protein degradation,inhibit the transcription levels of ARdownstream target genes,thereby suppressing ENZR cell proliferation and promoting apoptosis.This study provides a candidate compound for targeting ARfl/ARv7 to overcome ENZ resistance.
作者
韩正义
梁艳
曹鹏
尹震宇
李岩
HAN Zheng-yi;LIANG Yan;CAO Peng;YIN Zhen-yu;LI Yan(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;National Key Laboratory of Process Control and Intelligent Manufacturing Technology for Traditional Chinese Medicine Pharmaceutical Production,Nanjing University of Chinese Medicine,Nanjing 210023,China;Gulou Clinical Medical College,Nanjing University of Chinese Medicine,Nanjing 210008,China;Affiliated Jinling Hospital,Medical School of Nanjing University/General Hospital of Eastern Theater Command,Nanjing 210007,China)
出处
《药学学报》
北大核心
2025年第6期1739-1745,共7页
Acta Pharmaceutica Sinica
基金
国家自然科学基金青年科学基金项目(82304800)
江苏省高等学校基础科学(自然科学)研究面上项目(23KJB360014)
南京中医药大学校配套项目(XPT82304800)。
关键词
前列腺癌
耐药性
恩杂鲁胺
降解
雄激素受体
prostate cancer
drug resistance
enzalutamide
degradation
androgen receptor
作者简介
通讯作者:李岩,E-mail:yanli@njucm.edu.cn;通讯作者:尹震宇,zhenyuyin68@163.com。
