胆管癌相关成纤维细胞对人胆管癌细胞转移的影响  

Cancer associated fibroblasts promotes cancer metastasis in cholangiocarcinoma RBE cells

作　　者：武伟[1] 孟柠[2] 潘志坚[1] 李杭[1] 何桦波[1] 李国兰[3] Wu Wei;Meng Ning;Pan Zhijian;Li Hang;He Huabo;Li Guolan(Department of Gastrointestinal Hepatobiliary Surgery,the Affiliated Hospital of Hangzhou Normal Universityy Hangzhou 310015,China;Department of Breast Vascular Surgery,the Affiliated Hospital of HangzhouNormal University,Hangzhou 310015,China;Department,of Anesthesiology,the Affiliated Hospital ofHangzhou Normal University,Hangzhou 310015 China)

机构地区：[1]杭州师范大学附属医院胃肠肝胆外科,310015 [2]杭州师范大学附属医院甲乳血管疝外科,310015 [3]杭州师范大学附属医院麻醉科,310015

出　　处：《中华实验外科杂志》2019年第7期1211-1214,共4页Chinese Journal of Experimental Surgery

基　　金：浙江省医药卫生科技计划(2016KYB231、2016KYB229).

摘　　要：目的观察肿瘤相关成纤维细胞对胆管癌细胞RBE生长的影响.方法收集2016年6月至2017年5月来杭州师范大学附属医院诊断为临床原发性胆管癌患者的肿瘤组织分离到的肿瘤相关成纤维细胞(CAF),建立了CAF和癌旁成纤维细胞(PTF)的原代培养,并采用流式细胞术检测细胞内波形蛋白(Vimentin)和细胞角蛋白(Cytokeratin)的表达,蛋白质印迹法(Western blot)检测成纤维细胞标记蛋白成纤维细胞激活蛋白(FAPα)、成纤维特异性蛋白1(FSP1)和α-平滑肌肌动蛋白(α-SMA)表达.在CAF和RBE共培养的条件下,流式分析细胞增殖和凋亡,Transwell检测迁移、侵袭.应用SPSS 2.0软件统计分析.计量资料以均值±标准差(Mean±SD)表示,组间比较采用t检验.结果与PTF比较,CAF中FSP1、FAPα、α-SMA表达水平更高,Vimentin阳性率更高;在共培养条件下,CAF比PTF能显著提高胆管癌RBE细胞的增殖活性[(5.97±1.59)%]比[(1.59±0.35)%],促进RBE的侵袭[24 h,(86.76±6.11)%比(51.33±4.04)%]和迁移[24 h,(103.33±5.73)%比(87.00±4.85)%],差异有统计学意义(t=2.776,均P<0.05),但不影响RBE的凋亡,差异无统计学意义(t=2.776,P>0.05).结论胆管癌相关成纤维细胞CAF能促进RBE的侵袭和迁移,在胆管癌转移过程中起着重要作用.Objective Cancer-associated fibroblasts (CAF) have been reported to promote carcinogenesis and tumor progression. However, the role of cancer-associated fibroblasts on the metastasis of cholangiocarcinoma has not been clarified. The aim of this study is to investigate the effect of cancer-associated fibroblasts on progression of cholangiocarcinoma RBE cell in vitro. MethodsCancer-associated fibroblasts were isolated from the tumor tissue of patients. Then, we established primary cultures of CAF and peri-tumor fibroblasts (PTF) from patients with resected cholangiocarcinoma. Flow cytometric analysis, Western blotting and transwell assay were used in our study. ResultsThe results indicated a higher expression of fibroblast secreted protein-1, fibroblasts activating proteinα,α-Smooth muscle actin and Vimentin in CAF compared to that in PTF. Flow cytometric analysis and transwell assay indicated that conditioned medium from CAF cell cultures increased remarkably the proliferative activity of cholangiocarcinoma RBE cell (5.97±1.59)% vs.(1.59±0.35)%, promoted RBE invasion [24 h,(86.76±6.11)% vs.(51.33±4.04)%] and migration [24 h (103.33±5.73)% vs.(87.00±4.85)%] but had no influence on apoptosis of RBE. ConclusionOur study showed that CAF played an important role in reinforcing cholangiocarcinoma metastasis, suggesting as a therapeutic target in cholangiocarcinoma.

关 键 词：肿瘤相关成纤维细胞 增殖 侵袭 转移 胆管癌 

分 类 号：R735.8[医药卫生—肿瘤]