卵巢癌相关成纤维细胞通过上皮间质转化促进卵巢癌细胞SKOV3侵袭的研究

发布时间：2018-01-28 01:29

本文关键词： 卵巢肿瘤癌相关成纤维细胞上皮间质转化侵袭　出处：《肿瘤学杂志》2016年12期 　论文类型：期刊论文

【摘要】：[目的]研究癌相关成纤维细胞的条件培养基对卵巢癌细胞侵袭能力的影响及作用机理。[方法]原代培养人卵巢正常成纤维细胞(NOF)和癌相关成纤维细胞(CAF),并收取条件培养基NOFcm和CAFcm。用无血清的培养基DMEM、NOFcm和CAFcm分别处理SKOV3细胞,诱导SKOV3细胞发生上皮间质转化(EMT),通过形态观察和Western blot验证。形态上发生上皮间质转化(EMT)的SKOV3细胞,用transwell小室及三维培养实验检测细胞的侵袭能力的改变。[结果]CAF的条件培养基能在体外诱导卵巢癌细胞SKOV3在形态上发生改变:细胞连接疏松,部分细胞呈梭形,且明显下调E-cadherin蛋白表达量,明显上调Vimentin蛋白表达量。Transwell侵袭实验显示CAFcm处理后的SKOV3穿透matrigel胶的细胞数(284.00±32.86)相比于无血清的DMEM组(60.60±11.72)和NOFcm组(116.60±14.22),数量明显增多(P=0.000374和P=0.001258);三维培养实验中只有CAFcm组和NOFcm组能形成克隆球,而在无血清的DMEM组不能形成克隆球,CAFcm组(37.75±1.70)相比于NOFcm组(14.5+±1.29),形成的克隆球明显增大,向外侵袭能力增强,数量增多(P0.001)。[结论]卵巢癌CAF通过诱导卵巢癌细胞发生EMT,进而促进卵巢癌细胞的侵袭能力。
[Abstract]:[Objective] to study the effect and mechanism of conditioned medium of cancer-related fibroblasts on invasion ability of ovarian cancer cells. [Methods the primary culture of normal human ovarian fibroblasts (NOF) and carcinomatous fibroblasts (Cafs) were carried out, and conditioned media NOFcm and CAFcm.were used in serum-free medium DMEM. SKOV3 cells were treated with NOFcm and CAFcm, respectively, and SKOV3 cells were induced to develop epithelial mesenchymal transformation (EMT). Morphologic observation and Western blot test showed that SKOV3 cells were morphologically transformed into epithelial stroma. Transwell chamber and three-dimensional culture experiment were used to detect the change of cell invasion ability. [Results: the conditioned medium of CAF could induce the morphological change of ovarian cancer cell SKOV3 in vitro: the connection of the cells was loose, and some of the cells were fusiform. The expression of E-cadherin protein was down-regulated. Transwell invasion assay showed that the number of SKOV3 cells penetrating matrigel gel after CAFcm treatment was significantly increased (P < 0.05). 284.00 卤32.86) compared with serum-free DMEM group (60.60 卤11.72) and NOFcm group (116.60 卤14.22). The quantity of Pu 0.000374 and Pu 0.001258 were increased obviously. In three dimensional culture experiment, only CAFcm and NOFcm could form clone spheres, but in serum-free DMEM group, clone spheres could not be formed. Compared with NOFcm group, CAFcm group (37.75 卤1.70) significantly increased the formation of clone spheres and enhanced its outward invasion ability compared with NOFcm group (14.5 卤1.29). The quantity increased P0.001. [Conclusion: ovarian cancer CAF can promote the invasion of ovarian cancer cells by inducing EMTs.
【作者单位】：浙江省肿瘤医院;
【基金】：浙江省自然科学基金(LY14H160010)
【分类号】：R737.31
【正文快照】： 目前,肿瘤微环境的研究是肿瘤研究领域的热点。肿瘤微环境不仅在肿瘤细胞的恶性转化过程中发挥重要作用,而且在肿瘤细胞的转移复发阶段提供必要的支持[1,2]。肿瘤微环境中有多种细胞和细胞外基质,其中最主要的细胞是癌相关成纤维细胞专题报道(cancer-associated fibroblasts,C

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