基质成纤维细胞对卵巢癌细胞增殖能力的影响及相关分子机制  

Effect of stromal fibroblasts on proliferation capacity of ovarian cancer cells and related molecular mechanisms

作　　者：张亚[1] 杨英捷[1] ZHANG Ya;YANG Ying-jie(Department of Gynecological Oncologic Surgery,Guizhou Cancer Hospital,Guiyang 550000,China)

机构地区：[1]贵州省肿瘤医院妇瘤外科,贵阳市550000

出　　处：《广西医学》2020年第22期2956-2959,2964,共5页Guangxi Medical Journal

基　　金：贵州省科技合作计划(黔科合LH字7035号)。

摘　　要：目的分析基质成纤维细胞通过转化生长因子α(TGF-α)/表皮生长因子受体(EGFR)信号轴对卵巢癌细胞增殖的促进作用。方法(1)收集卵巢癌细胞SKOV-3和基质成纤维细胞WI38培养24 h后的条件培养基。使用四甲基偶氮唑蓝法检测经条件培养基处理24 h、48 h、72 h后卵巢癌细胞SKOV-3的增殖能力,使用实时荧光定量PCR和蛋白免疫印迹技术分别检测经条件培养基处理24 h后卵巢癌细胞SKOV-3中EGFR mRNA和蛋白的表达水平。采用酶联免疫吸附法检测条件培养基中上皮细胞生长因子(EGF)、白细胞介素6(IL-6)和TGF-α水平。(2)收集24例卵巢转移癌组织,运用免疫组织化学技术检测组织中TGF-α和EGFR的表达情况,并分析两者的相关性。结果与卵巢癌细胞系条件培养基相比,基质成纤维细胞条件培养基中TGF-α水平更高,经其处理后卵巢癌细胞SKOV-3的活力、EGFR mRNA和蛋白表达水平均更高(均P<0.05)。TGF-α和EGFR在卵巢转移癌组织中均呈高表达,且两者的阳性表达情况呈正相关(P<0.05)。结论基质成纤维细胞可能通过旁分泌TGF-α激活卵巢癌细胞中EGFR,从而促进卵巢癌细胞增殖,提示基质成纤维细胞可能在卵巢癌的转移定植过程中发挥重要作用。Objective To analyze the facilitation of stromal fibroblasts on the proliferation of ovarian cancer cells through transforming growth factorα(TGF-α)/epidermal growth factor receptor(EGFR)signal axis.Methods(1)The conditioned medium was collected after 24-hour culture of ovarian cancer cells SKOV-3 or stromal fibroblasts WI38.Methyl thiazolyl tetrazolium assay was used to determine the proliferation capacity of ovarian cancer cells SKOV-3 after 24,48 and 72 hours of treatment by conditioned medium;besides,quantitative real-time PCR and western blot were employed to detect the expression levels of EGFR mRNA and protein in ovarian cancer cells SKOV-3 after 24-hour treatment by conditioned medium,respectively.Enzyme-linked immunosorbent assay was used to determine the levels of epithelial growth factor(EGF),interleukin 6(IL-6)and TGF-αin conditioned medium.(2)Twenty-four specimens of ovarian metastatic cancer tissues were collected,then immunohistochemical techniques were used to detect the expression of TGF-α and EGFR in the tissues,and the correlation of the two indicators was analyzed.Results By comparison with conditioned medium of ovarian cancer cells,TGF-α level was higher in the conditioned medium of stromal fibroblasts,and ovarian cancer cells SKOV-3 yielded increased activity and possessed higher EGFR mRNA and protein expressions levels after treatment by conditioned medium of stromal fibroblasts(all P<0.05).TGF-α and EGFR were highly expressed in ovarian metastatic cancer tissues,and a positive correlation existed between the expression of the two indicators(P<0.05).Conclusion Stromal fibroblasts probably activate EGFR in ovarian cancer cells by paracrine of TGF-α,thus promoting the proliferation of ovarian cancer cells.This finding suggests that stromal fibroblasts may play an important role in the metastatic colonization of ovarian cancer.

关 键 词：卵巢癌 基质成纤维细胞 细胞增殖 转化生长因子Α 表皮生长因子受体 转移定植 

分 类 号：R737.31[医药卫生—肿瘤]