mTOR抑制剂西罗莫司及其衍生物对黑色素瘤细胞株A375的抑制作用  

Effectiveness of mTOR inhibitor Sirolimus and its derivates on melanoma cell line A375

作　　者：陈晓明[1] 蔡蕾 黄剑清 黄舒燕 郑从燊[1] 黄卫东 CHEN Xiaoming;CAI Lei;HUANG Jianqing;HUANG Shuyan;ZHENG Congshen;HUANG Weidong(Fujian Institute of Microbiology , Fuzhou, Fwin 350007, China)

机构地区：[1]福建省微生物研究所,福州350007 [2]福建省福州市皮肤病防治院

出　　处：《福建医药杂志》2019年第4期110-114,共5页Fujian Medical Journal

基　　金：福建省自然科学基金资助项目(2015J01362);福州市临床重点专科建设项目(201807111);福州市皮肤病与医学中心建设项目(2018080309)

摘　　要：目的比较临床上使用的mTOR抑制剂西罗莫司、依维莫司和替西罗莫司对黑色素瘤细胞株A375的抑制作用,为mTOR抑制剂治疗黑色素瘤提供参考。方法 SRB蛋白染色法检测药物对A375细胞增殖的影响,Annexin V-FITC/PI双染法检测药物对A375细胞凋亡的影响,PI标记检测药物对A375细胞周期的影响,Western blot检测药物对A375细胞mTOR及其下游蛋白磷酸化的影响。结果 mTOR抑制剂西罗莫司、依维莫司和替西罗莫司具有很强的抑制黑色素瘤细胞株A375增殖的作用,能够诱导A375细胞凋亡、阻滞细胞周期于G1期,并抑制A375细胞mTOR及其下游蛋白激酶4EBP1和p70S6K1的磷酸化。在这3个mTOR抑制剂中,依维莫司较西罗莫司和替西罗莫司表现出对A375更强的活性。结论依维莫司较替西罗莫司和西罗莫司在黑色素瘤的治疗上可能更有意义。Objective To compare the effectiveness of clinic mTOR inhibitors Sirolimus, Everolimus and Temsirolimus on melanoma cell line A375, which will provide insights into clinic application of mTOR inhibitors in melanoma. Methods Sulforhodamine B(SRB) assay was used to investigate the effectiveness of inhibitors on A375 cell proliferation. Annexin V-FITC/PI staining was performed to detect apoptosis. Propidium iodide(PI) staining was to analyze the influence of inhibitors on cell cycle. Western blot was applied to determine the regulation of mTOR as well as its downstream signaling. Results All of three mTOR inhibitors showed potent cell growth inhibition on A375. Mechanistically, they can promote A375 cell apoptosis, lead to cell cycle arrest at G1 phase by blocking mTOR and phosphorylation of its downstream targets 4 EBP1 and p70 S6 K1. Everolimus exhibited more potent activity against Sirolimus and Temsirolimus. Conclusion Everolimus is a more promising drug for melanoma treatment.

关 键 词：西罗莫司 依维莫司 替西罗莫司 黑色素瘤 

分 类 号：R739.5[医药卫生—肿瘤]