骨髓间充质干细胞对HT-29细胞所形成的裸鼠肿瘤的影响  

The effects of bone marrow derived mesenchymal stem cells on the tumor development in nude mice from HT-29 cells

作　　者：巩福星[1] 焦虎[1] 董平[1] 严笠[1] 尹艳花[1] 肖苒[1] 

机构地区：[1]中国医学科学院 北京协和医学院整形外科医院研究中心,北京100144

出　　处：《基础医学与临床》2012年第5期520-524,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30871433;31071305)

摘　　要：目的探讨骨髓间充质干细胞(BMSC)对结肠癌细胞HT-29所形成肿瘤生长和分化的影响。方法体内实验将裸鼠分为3组:HT-29组、BMSC组、BMSC+HT-29组。分别将相应细胞悬液注射到裸鼠皮下,测量形成肿块体积,7周后取材、称重并HE染色;real-time PCR和免疫组化检测上皮标志基因E-CADHERIN表达。体外实验将HT-29细胞与BMSC隔离共培养,观察HT-29细胞形态,RT-PCR检测上皮间质转化(EMT)相关基因表达。结果体内实验中BMSC组未见肿块形成,HT-29+BMSC组形成肿块质量和体积与HT-29组无明显差异,但肿块分化水平较低,E-CADHERIN表达水平较低。体外隔离共培养后HT-29细胞形态发生明显改变,EMT相关基因表达水平亦发生相应改变。结论 HT-29细胞形成肿瘤的分化程度可能与BMSC微环境诱导HT-29细胞发生EMT有关。Objective To study the effects of bone marrow derived mesenchymal stem cells on the growth and differentiation of the tumor developed from HT-29 cells.Methods In vivo,nude mice were divided into 3 groups-HT-29 group,BMSC group and HT-29＋BMSC group,then cell suspensions were subcutaneously injected into nude mice.Tumors were removed after 7 weeks.The volumes and weights of the tumors were measured and HE staining was performed.The expression of an epithelia marker-E-CADHERIN was detected using RT-PCR and immunohistochemistry staining.In vitro,HT-29 cells were co-cultured with BMSCs,the morphology of HT-29 cells was observed,and the expression of EMT related genes were detected using RT-PCR.Results In vivo,no tumor was observed in the BMSC group,and no significant difference was found in the volume and weight of tumors between the HT-29＋BMSC group and the HT-29 group.But the tumor cells in the HT-29＋BMSC group showed poorly differentiated and lower level expressed of E-CADHERIN compared with the HT-29 group.In vitro,after coculturing with BMSCs,morphological changes of HT-29 cells were observed and were consistent with the expression of EMT related genes.ConclusionsThe differentiation degree of HT-29 cells is correlated to EMT induced by BMSCs microenvironment.

关 键 词：骨髓间充质干细胞 HT-29细胞 肿瘤生长和分化 共培养 EMT 微环境 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]