间充质干细胞增强结直肠癌细胞可提高正常肠上皮细胞迁移能力并诱导上皮间质转化  

Mesenchymal stem cells promote the induction of colorectal cancer cells on normal intestinal epithelium

作　　者：胡赢 刘义[1,2] 韦成江 朱亚珍 赖浩 冯雁[3] 林源 HU Ying;LIU Yi;WEI Cheng-jiang;ZHU Ya-zhen;LAI Hao;FENG Yan;LIN Yuan(The Second Ward of Gastrointestinal Surgery,Guangxi Medical University Cancer Hospital,Nanning 530021,China;Colorectal Cancer Clinical Diagnosis and Treatment Center,Nanning 530021,China;Department of Research,Guangxi Medical University Cancer Hospital,Nanning 530021,China)

机构地区：[1]广西医科大学附属肿瘤医院胃肠二病区,广西南宁530021 [2]广西结直肠癌临床诊疗中心,广西南宁530021 [3]广西医科大学附属肿瘤医院实验研究部,广西南宁530021

出　　处：《海南医学院学报》2022年第9期669-675,684,共8页Journal of Hainan Medical University

基　　金：广西科技计划重点研发资助项目(AB18221086,AB16380202);广西自然科学基金青年基金项目(GXNSFBA138033)。

摘　　要：目的:体外构建肠癌细胞或间充质干细胞与正常结肠上皮细胞的共培养模型,模拟体外肿瘤微环境,探讨在体外共培养环境下肠癌细胞或间充质干细胞对正常结肠上皮细胞迁移能力和上皮-间质转化的影响。方法:用Transwell小室建立结肠癌细胞株(SW480)、人脐带间充质干细胞(hUC-MSCs)或二者同时与人正常结肠上皮细胞(NCM460)的体外共培养模型;按照共培养模型中上室或下室接种的细胞共分成4组:A:空白对照组(不接种细胞),B:只接种hUC-MSCs,C:只接种SW480,D:混合接种hUC-MSCs和SW480;另一侧均接种NCM460细胞,上下室接种细胞的数目比例均为1∶1。观察共培养后NCM460的形态学改变,并通过划痕实验和Transwell实验检测NCM460迁移能力变化;同时用Westernblot检测NCM460共培养后的上皮-间质转化标志物改变。结果:各实验组NCM460细胞形态学上未发现显著改变;Transwell迁移实验结果表明,与hUC-MSC或SW480共培养时,NCM460的迁移能力有增强的趋势,Vimentin表达显著升高(P<0.05),E-cadherin蛋白的表达与划痕愈合能力改变不显著。当同时与MSCs和SW480共培养时,与空白对照组相比,NCM460细胞划痕愈合速度和Transwell迁移能力均显著增强,同时Vimentin蛋白表达显著上调,E-cadherin蛋白显著下调,差异具有统计学意义(P<0.05)。结论:结肠癌细胞可以提高正常结肠上皮细胞迁移能力和上皮间质转化标志物表达,间充质干细胞的存在对这一现象有增强作用。Objective:To establish a co-culture model of colon cancer cells or mesenchymal stem cells and normal colon epithelial cells in vitro to simulate the tumor microenvironment in vitro,and to investigate the effect of colon cancer cells or mesen⁃chymal stem cells on the migration ability and epithelial-mesenchymal transformation of normal colon epithelial cells in the co-cul⁃ture environment.Methods:In vitro co-culture model of colon cancer cell line SW480 or human umbilical cord mesenchymal stem cells(hUC-MSCs)with human normal colon epithelial cells(NCM460)was established by Transwell cell.Morphological chang⁃es of NCM460 after co-culture were evaluated by microscopic observation,and migration ability of NCM460 was measured by scratch experiment and Transwell experiment.Meanwhile,Western blot was used to detect the changes of epithelial-mesenchymal transition(EMT)markers after co-culture of NCM460.Results:There was no significant change in cell morphology of NCM460 in all experimental groups.Transwell migration results showed that when hUC-MSC or SW480 were co-cultured with NCM460,there was a trend of enhancement of the migration ability of NCM460,and the expression of Vimentin protein was significantly up-regulated to a certain extent(P<0.05),but E-cadherin protein expression and scratch healing ability were not significantly changed(P>0.05).When MSCs and SW480 were present at the same time,both scratch test and Transwell test showed that NCM460's scratch healing ability and migration ability were significantly enhanced(P<0.05),while Vimentin protein expression was significantly up-regulated and E-cadherin protein was significantly down-regulated,with statistically significant differences(P<0.05).Conclusion:Colon cancer cells can increase the migration ability of normal colonic epithelial cells and the expression of epithelial-mesenchymal transition markers.Mesenchymal stem cells may promote or act synergistically in this process.

关 键 词：肿瘤微环境 上皮-间质转化 恶性肿瘤 间充质干细胞 

分 类 号：R735.34[医药卫生—肿瘤]