Stathmin表达与食管鳞癌细胞对紫杉醇敏感性的机制研究  

Expression of stathmin and its influence on taxol sensitivity in esophageal squamous cell carcinoma

作　　者：韩改净 颜露[1] 牛芳斐 刘芳 周兰萍[1] 赵晓航[1] 许杨[1] 

机构地区：[1]国家癌症中心/中国医学科学院北京协和医学院肿瘤医院,北京理学硕士研究生100021

出　　处：《医学研究生学报》2016年第7期677-682,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81372385);国家高技术研究发展计划(2012AA02A503)

摘　　要：目的微管解聚蛋白Stathmin在食管鳞癌中的表达水平显著升高,而紫杉醇是靶向微管的临床食管癌常用化疗药物,文中探讨Stathmin高表达对食管鳞癌细胞紫杉醇敏感性的影响。方法以食管鳞癌KYSE 510和KYSE 170细胞为研究对象,构建过表达STMN1基因的食管鳞癌细胞系KYSE 510-Stathmin、KYSE 170-Stathmin和对照细胞KYSE 510-Control、KYSE 170-Control;过表达Stathmin的设定为Stathmin过表达组,同时以转染空载体筛选获得的单克隆细胞KYSE 510-Control做为对照组。MTT实验和集落形成实验比较实验组细胞与对照组细胞对紫杉醇敏感性;流式分析检测紫杉醇处理后KYSE 510-Stathmin与KYSE 510-Control细胞凋亡情况;Western blot检测凋亡和自噬相关分子的变化情况。结果在KYSE510和KYSE 170细胞中外源表达STMN1基因,Stathmin蛋白水平显著高于对照细胞(P<0.01);紫杉醇浓度为50、100、250nmol/L处理24 h,KYSE 510-Control的抑制率均高于KYSE 510-Stathmin细胞(P<0.01);250 nmol/L的紫杉醇处理24、48、60h,KYSE 510-Control细胞抑制率均高于KYSE 510-Stathmin(P<0.01),在KYSE 170-Stathmin细胞中获得一致性结果;紫杉醇处理后,KYSE 510-Stathmin集落数量显著高于KYSE 510-Control(P<0.01);流式细胞仪检测KYSE 510-STMN1细胞凋亡比例低于KYSE 510-Control细胞[(11.90±0.78)%vs(29.63±3.26)%,P<0.05]。Western blot提示凋亡相关分子Caspase8和Caspase9活化片段增多。结论过表达Stathmin影响食管鳞癌细胞对微管相关化疗药物紫杉醇的敏感性。Objective Stathmin,a microtubule-destabilizing protein,has high expression in esophageal squamous cell carcinoma（ESCC）,while taxol is a common chemotherapy microtubule-targeted drug for esophageal cancer.This study aimed to investigate the impact of stathmin expression and its influence on taxol sensitivity in ESCC.Methods We established 2 cell models with STMN1 gene overexpression in KYSE 510 and KYSE 170 cell lines,including KYSE 510-Stathmin,KYSE 170-Stathmin,KYSE 510-Control and KYSE 170-Control.MTT assay and colony formation were applied to compare the taxol sensitivity between experimental group and control group.Flow cytometry was used to measure the apoptosis of KYSE 510-Stathmin and KYSE 510-Control after taxol treatment.Western blot was used to test the changes of related factors to apoptosis and autophagy.Results（1） Stathmin protein expressions in KYSE 510-Stathmin and KYSE 170-Stathmin cells were higher than those of control cells（P〈0.01）.（2） The percentages of inhibition were significantly decreased in KYSE 510-Stathmin and KYSE 170-Stathmin cells 24 h after 50,100,250 nmol/L taxol treatment compared with KYSE 510-Stathmin cells（P〈0.01）.（3） The percentages of inhibition were significantly reduced in KYSE 510-Stathmin and KYSE 170-Stathmin cells after 250 nM taxol treatment for 24,48,60 h（P〈0.01）.（4）After taxol treatment,the number of colony formation in KYSE 510-Stathmin cells was higher compared with KYSE 510-Control cells（P〈0.01）.（5）The percentage of cell apoptosis in KYSE 510-Stathmin was significantly lower than that of KYSE 510-Control cells by flow cytometry（11.90% ± 0.78% vs 29.63% ± 3.26%,P〈0.05）.Western blot showed the apoptosis of associated proteins such as the activation of Caspase8 and Caspas9.Conclusion The result indicates that overexpression of stathmin inhibits taxol sensitivity in ESCC cell lines.

关 键 词：STATHMIN 食管鳞状细胞癌 紫杉醇 

分 类 号：R735[医药卫生—肿瘤]