摘要
目的:探讨IL-6诱导卵巢癌细胞对他莫西芬(tamoxifen,TAM)耐药的分子机制。方法:构建内源性过表达IL-6的人卵巢癌A2780细胞系和内源性抑制IL-6表达的人卵巢癌CAOV-3细胞,50 ng/ml外源性IL-6预处理A2780细胞(A2780/pre IL-6细胞),Western blotting检测内/外源IL-6对卵巢癌细胞ERαSer167位磷酸化水平的影响;IL-6与PI3K抑制剂Wortmannin单独或联合作用于A2780细胞,Western blotting检测其对A2780细胞Akt磷酸化和ERα磷酸化的影响;MTT法检测Wortmannin和内/外源IL-6对A2780细胞TAM敏感性的影响;荧光素酶报告基因检测卵巢癌细胞ERα的转录活性,并分析其可能涉及的信号通路。结果:外源性及内源性过表达IL-6可明显促进A2780细胞ERαSer167位点磷酸化水平(均P<0.01),而内源性抑制IL-6表达则可降低CAOV-3细胞ERαSer167位点的磷酸化水平(P<0.01);Wortmannin可阻断IL-6诱导的A2780细胞对TAM的耐药及ERα的磷酸化(P<0.05);IL-6可促进细胞ERα的转录活性(P<0.01),而Wortmannin并不能阻断IL-6对ERα的转录活性的影响(P>0.05)。结论:IL-6可经PI3K/Akt通路引起ERα磷酸化从而活化ER信号通路,进而诱导卵巢癌细胞对TAM耐药。
Objective：This study aimed to explore the mechanism of tamoxifen （TAM） resistance caused by IL- 6 in ovarian cancer cells. Methods： Human ovarian cancer A2780 cell line that endogenously over expressing IL 6 and human ovarian cancer CAOV 3 cell line that exogenously depleting IL 6 were constructed; exogenous IL- 6 （50 ng/ml） were used for pretreatment of A2780 cells （A2780/perIL- 6 cells）, and Western blotting was used to detect the effect of endogenous/exogenous IL- 6 on the phosphorylation level of ERα Ser167 in ovarian cancer cells; IL- 6 and/or Wortmannin （PI3K inhibitor） were used to treat A2780 cells and western blotting was used to detect their effect on the phosphorylation of Akt and ERα; MTT assay was used to detect the effect of Wortmannin and endogenous/exogenous IL- 6 on the sensitivity of A2780 cells to TAM; luciferase reporter assay was performed to detect transcription activity of ERα in ovarian cancer cells, and to explore the possible signaling pathway. Results： Both exogenous and endogenous over expression of IL- 6 could obviously increase the level of ERα Ser167 phosphorylation in A2780 cells （all P〈0.01）, while endogenous depletion of IL- 6 could reduce the level of ERα Ser167 phosphorylation in CAOV 3 cells （P〈0.01）. It also found that wortmannin （PI3K inhibitor） could significantly antagonize IL 6 induced TAM resistance and phosphorylation of ERα Ser167. IL 6 promoted ERa transcription activity, while this activation was not blocked by the PI3K specific inhibitor wortmannin. Conclusion： These results indicate that IL 6 could induce ERa phosphorylation by triggering PI3K/Akt signaling pathway to activate the ER pathway, and thereby induce the resistance of ovarian cancer cells to TAM.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2017年第6期601-607,共7页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.81572852
No.81502256)
天津市自然科学基金资助项目(No.12JCZDJC26300)
武警后勤学院科学技术研究资助项目(No.2015ZXKF05
No.WHB201404
No.WHB201505)~~
