HSP90α抑制剂17-AAG抑制甲状腺癌细胞增殖和迁移机制探讨  

17-AAG of Hsp90 alpha inhibitor effects on proliferation and migration of thyroid cancer cell

作　　者：李云芳 董婷婷 孔霞 张国安 王业全 侯森 庞潜潜 崔文 LI Yun-fang;DONG Ting-ting;KONG Xia;ZHANG Guo-an;WANG Ye-quan;HOU Sen;PANG Qian-qian;CUI Wen(School of Medicine and Life Sciences,University of Jinan,Shandong Academy of Medical Sciences,Jinan 250062,P.R.China;Medical College of Qingdao University,Qingdao 266000,P.R.China;Department of Pharmacy,Jining First People's Hospital,Jining 272111,P.R.China;Institute of Forensic Medicine and Laboratory Medicine of Jining Medical University,Jining 272067,P.R.China)

机构地区：[1]济南大学山东省医学科学院医学与生命科学学院,山东济南250062 [2]青岛大学医学部,山东青岛266000 [3]济宁市第一人民医院药学部,山东济宁272111 [4]济宁医学院法医学与医学检验学院,山东济宁272067

出　　处：《中华肿瘤防治杂志》2019年第1期6-11,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：济宁医学院青年教师科研扶持基金(JY2017FY002);济宁医学院青年教师科研扶持基金(JY2017FY001);济宁医学院大学生创新训练计划(cx2016034)

摘　　要：目的在体内,热休克蛋白90α(heat shock protein 90α,HSP90α)对肿瘤的发生发展起着重要的作用。HSP90α抑制剂在体内可阻断肿瘤赖以生存的信号通路网络。本研究探讨HSP90α特异性抑制剂烯丙胺基-17-去甲氧基格尔德(17-allyl-amino-17-demethoxygeldanamycin,17-AAG)对甲状腺癌细胞增殖和迁移的影响及相关信号分子的调控机制。方法体外培养甲状腺乳头状癌细胞TPC-1和甲状腺未分化癌细胞TAK,分别给予0.001～10.000μmol/L浓度的17-AAG作用24和48h。MTT实验、划痕实验观察细胞生长和迁移情况。蛋白质印迹法检测细胞HSP90α及信号传导及转录激活因子(signal transducers and activators of transcription,STAT3)、丝氨酸/苏氨酸蛋白激酶(serine/threonine protein kinase,AKT)、细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)相关信号蛋白的表达及磷酸化水平变化。结果与未处理的细胞相比,经0.001～10.000μmol/L浓度的17-AAG作用24和48h的TPC-1和TAK细胞增殖能力降低,且有明显的时间和剂量依赖性。TPC-1(F=7.326,P=0.005)和TAK细胞(F=22.770,P<0.001)浓度组间比较差异有统计学意义,时间组间比较差异有统计学意义,F值分别为1 048.000和663.800,均P<0.001。与未处理的细胞相比,经0.1和1.0μmol/L浓度的17-AAG作用24h的TPC-1和TAK细胞迁移能力降低,各浓度组间比较差异有统计学意义,F=1.228,P<0.005;两细胞株间比较差异无统计学意义,F=0.365,P=0.578。经0.1和1.0μmol/L浓度的17-AAG作用24h的TPC-1和TAK细胞以及STAT3、AKT、ERK蛋白磷酸化水平降低,却对STAT3和ERK总蛋白表达影响差异无统计学意义,P>0.05。结论 17-AAG通过下调甲状腺癌细胞HSP90α相关信号蛋白STAT3/AKT/ERK磷酸化水平,从而抑制甲状腺癌细胞的增殖和迁移能力。OBJECTIVE Heat shock protein 90 alpha plays an important role in tumor development,HSP90αinhibitor can block tumor survival signaling pathway in the body.This study was designed to explore the effects of HSP90αinhibitor 17-AAG on the proliferation and migration of thyroid cancer cell and the underlying signaling mechanisms.METHODS Papillary thyroid cancer cell TPC-1 and undifferentiated thyroid cancer cell TAK were treated with 17-AAG,which was given the specific dose of 0.001-10.000μmol/L for 24 hand 48 hrespectively,MTT experiment and scratching assay were used to detect cell growth and migration.Western blot was used to detected HSP90αand STAT3/AKT/ERK signaling protein expression and phosphorylation level changes.RESULTS Compared with untreated cells,cell proliferation capacity was reduced after 24 hand 48 htreated with the concentration of 0.001-10.000μmol/L of 17-AAG,showing a significant time-and dose-dependent manner,comparison between more concentration groups(F=7.326,P=0.005;F=22.770,P<0.001),and the comparison between two-time groups(F=1 048.000,P<0.001;F=663.800,P<0.001)was statistically significant.TPC-1 and TAK cells migration capacities were reduced in the dose of 0.1μmol/L and1.0μmol/L of 17-AAG after 24 hcompared with untreated cells.Comparison between more concentration groups(F=1.228,P<0.005)was statistically significant,and comparison between two cells(F=0.365,P=0.578)was not statistically significant.Meanwhile,the phosphorylation levels of STAT3/AKT/ERK protein were reduced treated with 0.1 and1.0μmol/L of 17-AAG after 24 hin TPC-1 and TAK cells,but there was no statistical significance of general protein of STAT3 and ERK.CONCLUSION 17-AAG inhibits the proliferation and migration of thyroid cancer cell by reducing the phosphorylation of STAT3/AKT/ERK signaling proteins.

关 键 词：甲状腺癌 TPC-1 TAK HSP90Α 17-AAG STAT3/AKT/ERK 

分 类 号：R736.1[医药卫生—肿瘤]