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  

Ovarian Cancer-associated Fibroblasts Promotes Invasion of Ovarian Cancer SKOV3 Cell Lines Through Epithelial-mesenchymal Transition

作　　者：高雯[1] 朱滔[1] 

机构地区：[1]浙江省肿瘤医院,浙江杭州310022

出　　处：《肿瘤学杂志》2016年第12期988-992,共5页Journal of Chinese Oncology

基　　金：浙江省自然科学基金(LY14H160010)

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(P<0.001)。[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。[Objective] To explore the effects of cancer-associated fibroblasts（CAF） on invasion ability of epithelial ovarian cancer cells（EOC）. [Methods] Normal human ovarian fibroblasts（NOF）and CAF were primarily cultured and the conditioned mediums（CM） of normal human ovarian fibroblast（NOFcm） and CAF（CAFcm） were collected. Ovarian cancer SKOV3 cells were treated with serum free medium（DMEM）,NOFcm or CAFcm to induce epithelial-mesenchymal transition（EMT）.EMT markers E-cadherin and Vimentin were detected by Western blot after SKOV3 cells were treated. Cell invasion ability was measured by Transwell chamber and three-dimensional cell culture. [Results] After treatment with CAFcms,SKOV3 cells had loose connections,part of the cells was spindle shaped; the E-cadherin protein level was down-regulated and Vimentin was upregulated. Transwell assay showed that the migration and invasion of SKOV3 cells treated with CAFcm（284.00±32.86）were significantly increased,compared with NOFcm group（116.60 ±14.22）and DMEM group（60.60±11.72）（P=0.000374 and P=0.001258,respectively）. In three-dimensional culture models,clone spheres were formed in CAFcm group and in NOFcm group,but not in DMEM group. The clone spheres in CAFcm group were bigger and more aggressive than those in NOFcm group; and the number of clone spheres in CAFcm group（37.75±1.70） was significantly increased compared with NOFcm group（14.50±1.29）（P〈0.001）. [Conclusion] CAF induces the EMT of ovarian cancer cells,which thereby promotes the invasiveness of ovarian cancer cells in vitro.

关 键 词：卵巢肿瘤 癌相关成纤维细胞 上皮间质转化 侵袭 

分 类 号：R737.31[医药卫生—肿瘤]