中文摘要：

目的检测卵巢癌患者腹水中肿瘤相关成纤维细胞（CAF）含量,并探讨其对卵巢癌肿瘤细胞侵袭能力的影响。方法从20例卵巢癌腹水中分离细胞后,流式检测其中α-SMA细胞阳性率并分为高低两群。在TCGA数据库557例卵巢癌标本数据库中验证α-SMA与上皮间质转化（EMT）相关的转录因子（SNAI1、TWIST1、ZEB1）的相关性;在mRNA和蛋白水平比较以上两群中肿瘤细胞上皮间质转化程度;磁珠分选成纤维细胞贴壁后α-SMA免疫荧光观察其形态;Western Blot验证SKOV3细胞系与人胚肺成纤维细胞系MRC5体外悬浮共培养后EMT程度的改变,Transwell和系膜清除试验验证在悬浮共培养状态下MRC5对SKOV3侵袭能力的影响。结果卵巢癌腹水中α-SMA阳性细胞占腹水细胞（34.40±18.67）%;TCGA数据库中α-SMA与上皮间质转化相关的转录因子显著正相关（r值分别为0.347 0、0.526 2和0.700 2,P〈0.01）;实时定量PCR（qRT-PCR）及Western Blot验证α-SMA-H组中肿瘤细胞EMT程度较α-SMA-L组更高;腹水中成纤维细胞贴壁免疫荧光成高表达α-SMA的间质样细胞形态;体外悬浮共培养后,MRC5促进SKOV3的EMT和系膜清除能力。结论卵巢癌腹水中存在CAF并可以促进卵巢癌腹水中肿瘤细胞EMT和侵袭能力。

英文摘要：

Objective To detect the content of cancer-associated fibroblasts（ CAF） in ascites of ovarian cancer patients,explore the impact on invasion of ovarian cancer cells. Methods After isolation of cells from malignant ascites of 20 ovarian cancer patients,flow cytometry was performed to detect the percentage of CAF with positive α-SMA,then the cells were divided into high α-SMA group and lowα-SMA group.; TCGA database was used to analyze the correlation between α-SMA and the relative genes of epithelial-mesenchymal transition（ EMT）（ SNAI1,TWIST1,ZEB1） among 557 ovarian cancers. Another verification was taken in mRNA and protein level to find out the differences between tumor cells between low and high α-SMA group on the extent of EMT. Magnetic sorting was used to purify fibroblasts from malignant ascites to observe the cell morphology by immunofluorescence of α-SMA. Western Blot was taken to confirm the change of EMT degree after co-culture of SKOV3 cell line and MRC5. Transwell and mesangial clearance assay were employed to analyze the effect of MRC5 on invasion ability of SKOV3 cells. Results The percentage of α-SMA positive cells in malignant ascites was（ 34. 40±18. 67） %.The mRNA levels of α-SMA and EMT associated genes（ SNAI1,TWIST1,ZEB1） were significantly positively correlated with TCGA（ r =0. 347 0,0. 526 2,0. 700 2,P〈0. 01）; tumor cells isolated from high α-SMA group got more EMT phenotypes than that from low α-SMA group. Fibroblasts shared similar morphology with high α-SMA expression cells. After in vitro suspension co-culture assay,MRC5 could facilitate the process of EMT and mesangial clearance of SKOV3 cells. Conclusion Cells in ascites of ovarian cancer patients contain CAF and promote EMT and mesangial clearance of ovarian cancer cells.