现代妇产科进展2016，Vol.25Issue(1)：1-5,5.DOI:10.13283/j.cnki.xdfckjz.2016.01.001
二甲双胍通过下调IL-6抑制卵巢癌肿瘤相关成纤维细胞活性的研究
Metformin suppresses the activity of ovarian cancer associated fibroblast through the ;downregulation of IL-6
摘要
Abstract
Objective:To explore the influence of metformin on ovarian cancer associat-ed fibroblast ( CAF ) and the involved mechanism. Methods:RT-PCR was done to detect the effect of metformin on mRNA levels of inflammatory cytokines ( IL-6, OPN, IL-1b, COX-2, Cyr61) in SKOV3 and primary CAFs. Activated MRC5 was gotten from that cultured with SK-OV3-CM for 7 to 10 days and was named MRC5-CAF. MRC5-CAF and primary CAFs were i-dentified by immunofluorescence ofα-SMA. Then the effect of metformin on the aforementioned cytokines was detected in MRC5-CAF using immunoblotting. Online TCGA data base ( http://www. cbioportal. org ) was employed to analyze the mRNA correlation between IL-6 and the CAFs characteristic moleculars among 557 ovarian cancers. Immunofluorescence was adopted to further verify the impact of metformin on IL-6 andα-SMA expressions of activated MRC5 . More-over,we performed the same treatment in MRC5-CAF to validate the influence of metformin or IL-6 on CAF characteristic genes. In vitro proliferation assay and transwell were employed to ex-amine the effects of metformin or IL-6 on CAFs' support for cancer cells' proliferaton and inva-sion. Collagen contraction experiment was done to check the alteration of CAFs in contraction of the ECM substrate under the influence of metformin or IL-6 . Result:Metformin was noted to re-duce the inflammatory cytokines expression,especially IL-6 and it could attenuate the character-istic a-SMA level in CAFs. The mRNA levels of IL-6 and CAF associated genes ( FAP, PDG-FRB,FN1,COLL6A6)were dramatically positively-correlated in TCGA datasets(P<0. 0001). Metformin and IL-6 co-stimulation regulated the activation of STAT3 signaling by IL-6 and its subsequent effect on CAF properties comparing to IL-6 group. It was confirmed that metformin could impair the capacities of CAF in promoting the proliferation,invasion and collagen contrac-tion induced by IL-6 . Conclusions:IL-6 is positively correlated with CAFs charactersitc genes and probably sustain the CAFs property in ovarian cancer. Metformin can probably downregulate IL-6 expression and the subsequent STAT3 activation mediated maintainence of CAFs property, thus diminishing the ability of CAFs in supporting of ovarian cancer cells' proliferation and in-vasion.关键词
卵巢癌/肿瘤相关成纤维细胞/二甲双胍/增殖/侵袭/胶原回缩Key words
Ovarian cancer/CAFs/Metformin/Proliferation/Invasion/Collagen con-traction分类
医药卫生引用本文复制引用
徐森,杨宗元,靳平,张陶然,高庆蕾..二甲双胍通过下调IL-6抑制卵巢癌肿瘤相关成纤维细胞活性的研究[J].现代妇产科进展,2016,25(1):1-5,5.基金项目
国家自然科学基金资助项目(No:81372801) （No:81372801）
