姜黄素通过阻滞MAPK/ERK通路抑制人子宫内膜癌HEC-1-B细胞的侵袭转移  

Curcumin inhibits the invasion and metastasis of endometrial carcinoma HEC-1-B cells by blocking MPAK / ERK signaling pathway

作　　者：陈茜[1] 陈丽娟[1] 党媛媛[1] 李牧[1] 吴晓玲[1] 

机构地区：[1]西安交通大学第二附属医院妇产科,西安710004

出　　处：《山西医科大学学报》2015年第8期762-768,共7页Journal of Shanxi Medical University

基　　金：陕西省科学技术研究发展计划基金资助项目(2014K11-01-01-18)

摘　　要：目的阐明MAPK/ERK信号传导途径在姜黄素抑制子宫内膜癌细胞侵袭转移中的作用及其机制。方法不同浓度(0,10,20和30μmol/L)姜黄素干预HEC-1B细胞后,用Western blot检测细胞中ERK1/2、p-ERK1/2、MEK1、p-MEK1及上皮间质转化(epithelial-mesenehymal transition,EMT)相关蛋白(E-cadhrein、Vimentin、Twist和Snail)的表达;利用pc DNA3.1(+)-MEK1表达载体和pc DNA3.1(+)空载体稳定转染细胞后过表达MEK1;利用Transwell侵袭实验检测姜黄素及ERK抑制剂U0126对HEC-1-B细胞的侵袭能力的效应;利用real-time PCR法检测姜黄素对EMT相关分子(CDH1、CDH2、FN1、Snail 1、Twist 1、Vimentin、ZEB1和ZEB2)的mRNA水平的影响。结果与空白对照组相比,姜黄素(≥20μmol/L)能够显著抑制HEC-1-B细胞中ERK1/2的活化,且呈浓度依赖性(P<0.01)。外源性过表达MEK1可以削弱姜黄素对HEC-1-B细胞的抗侵袭作用。单独使用U0126或姜黄素均可显著降低HEC-1-B细胞侵袭力(P<0.01或P<0.05)。此外,联合使用U1026能够增强姜黄素的侵袭抑制作用。进一步的Western blot和real-time PCR结果表明,姜黄素通过阻滞ERK信号转导通路,抑制EMT的发生。结论姜黄素通过阻滞MAPK/ERK信号转导通路抑制子宫内膜癌细胞的侵袭转移,这将为临床肿瘤转移和复发的治疗提供理论依据。Objective To elucidate the role and mechanism of MAPK/ERK signaling pathway in anti-invasive and mestastatic effect of curcumin in human endometrial carcinoma cells.Methods HEC-1-B cells were cultured in vitro and treated with different concentrations（0,10,20 and 30 μmol/L） of curcumin for 24 h.The protein levels of ERK1/2,p-ERK1/2,MEK1,p-MEK1 and EMT-related genes（E-cadhrein,Vimentin,Twist and Snail）were detected by Western blot.Exogenous MEK1 was stably transfected into HEC-1-B cells using pcDNA3.1 （＋）-MEK1 expression plasmid and pcDNA3.1 （＋）empty vector.Transwell chamber was used to evaluate the effects of curcumin and U0126 on invasion of HEC-1-B cells.The mRNA levels of EMT-related markers（CDH1,CDH2,FN1,Snail 1,Twist 1 Vimentin,ZEB1 and ZEB2） were measured in HEC-1-B cells treated with curcumin by real-time PCR.Results Curcumin（≥20 μmol/L） significantly inhibited the activation of ERK1/2 in HEC-1-B cells in a concentration-dependent manner（P ＜ 0.01）.Ectopic MEK1 expression relieved the inhibitory effect of curcumin on cell invasion.U1026 or curcumin alone significantly decreased the invasion of HEC-1-B cells（P ＜ 0.01 or P ＜ 0.05）.Curcumin combined with U1026 enhanced the anti-metastatic effect induced by curcumin.Western blot analysis and real-time PCR showed that curcumin inhibited EMT progression by suppression of ERK signal pathway.Conclusion These findings suggest that curcumin could inhibit the invasion and mestastasis of endometrial cancer cells by blocking the MAPK/ERK signal transduction pathway,which would provide a theoretical basis for the clinical treatment of tumor metastasis and recurrence.

关 键 词：姜黄素 子宫内膜癌 侵袭 ERK信号通路 EMT 

分 类 号：R737.33[医药卫生—肿瘤]