前列腺癌微环境中癌相关成纤维细胞对细胞缝隙连接通讯的影响  

Effect of cancer-associated fibroblasts under the prostate cancer-related microenvironment on the gap junctional intercellular communication

作　　者：陈东宁[1] 陈烨辉 蔡海[1] 孙雄林[1] 林云知 李晓东[1] 林婷婷[1] 魏勇[1] 郑清水[1] 薛学义[1] 许宁[1] Chen Dongning;Chen Yehui;Cai Hai;Sun Xionglin;Lin Yunzhi;Li Xiaodong;Lin Tingting;Wei Yong;Zheng Qingshui;Xue Xueyi;Xu Ning(Department of Urology,the First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China)

机构地区：[1]福建医科大学附属第一医院泌尿外科,福州350005

出　　处：《中华实验外科杂志》2020年第8期1431-1433,共3页Chinese Journal of Experimental Surgery

基　　金：福建医科大学启航基金(2017XQ1083);福建省教育厅中青年教师教育科研项目(JT180194);福建省科技创新联合资金项目(2017Y9093)。

摘　　要：目的观察前列腺癌微环境中癌相关成纤维细胞(CAFs)对细胞缝隙连接通讯(GJIC)的影响。方法取前列腺癌组织分离培养及鉴定CAFs,分别将(0、0.25、0.50、0.75、1.00 g/L)浓度的CAFs培养液与处于对数生长期的前列腺癌PC-3细胞(购自中国科学院上海生命科学研究院细胞资源中心)共同培养,采用荧光漂白恢复技术检测各组前列腺癌PC-3细胞缝隙连接(GJ)功能,蛋白质印迹法(Western blot)检测各组细胞间隙连接蛋白43(Cx43)蛋白的表达。组间比较采用方差分析。结果成功分离培养CAFs,前列腺癌PC-3细胞与不同浓度CAFs(0.00、0.25、0.50、0.75、1.00 g/L)共培养后,细胞内荧光恢复程度逐渐减弱[(49.40±5.42)%、(45.36±3.23)%、(36.57±3.95)%、(29.18±5.51)%、(21.81±4.58)%,F=18.067,P<0.05],细胞GJIC水平逐渐减弱,呈明显的剂量-效应依赖关系,差异有统计学意义(t=1.109、3.313、4.531、6.734,P<0.05)。同时,各组CAFs中的Cx43蛋白的表达水平分别为0.723±0.065、0.641±0.084、0.425±0.078、0.314±0.067、0.267±0.059。随着共培养CAFs浓度的增加,细胞内Cx43蛋白表达水平反而逐渐降低,差异有统计学意义(t=1.337、5.084、7.589、8.997,P<0.05)。结论CAFs可下调前列腺癌PC-3细胞Cx43蛋白的表达水平,抑制前列腺癌PC3细胞的细胞间缝隙连接功能。Objective To investigate the role of connexin 43(Cx43)and its derived gap junctional intercellular communication(GJIC)in the interplay between prostate cancer(PC)cells and cancer-associated fibroblasts(CAFs).Methods CAFs were separated from the prostate cancer issues and authenticated by Western blotting.Then different concentrations of CAFs(0,0.25,0.50,0.75,1.00 g/L)and PC-3 cells were co-cultured with direct contact.The function of GJIC in different groups was measured by the fluorescence recovery after photobleaching.Meanwhile,the expression of Cx43 in different groups was tested by Western blotting.Results CAFs were isolated from the prostate cancer samples and cultivated successfully.The fluorescence recovery rate[(49.40±5.42)%,(45.36±3.23)%,(36.57±3.95)%,(29.18±5.51)%,(21.81±4.58)%,F=18.067,P<0.05],and the expression of Cx43 were decreased with the increase of the concentration of CAFs(0.00,0.25,0.50,0.75,1.00 g/L)nutrient solution when co-cultured with PC-3 cells(P<0.05).The function of the GJIC became gradually less potent,which showed a dose-effect dependent relation.And there were significant difference(t=1.109,3.313,4.531,6.734,P<0.05)among these groups.Simultaneously,the expression level of Cx45 protein among different concentration groups of CAFs(0.00,0.25,0.50,0.75,1.00 g/L)was 0.723±0.065,0.641±0.084,0.425±0.078,0.314±0.067,0.267±0.059 respectively.The expression level of Cx43 in PC-3 cells showed a inverse relationship with the concentration of CAFs(t=1.337,5.084,7.589,8.997,P<0.05).Conclusion CAFs decreased the expression of CX43 in the PC-3 cells and inhibitited the function of the GJIC between prostate cancer cells.

关 键 词：前列腺癌 癌相关成纤维细胞 细胞缝隙连接通讯 

分 类 号：R737.25[医药卫生—肿瘤]