人脐带间充质干细胞对胶质瘤细胞株U87上皮间质转化及皮下荷瘤能力的影响  

Effects of human umbilical cord mesenchymal stem cell on epithelial-mesenchymal transition and growth of Glioma cell line U87

作　　者：刘高米洋[1] 和法莲 陈慧芬[2] 陆容[3] 李自安[1] 何洁[1] 赵晶[1] 潘兴华[1] Liu Gaomiyang He Falian Chen Huifen Lu Rong Li Zi＇an He Jie Zhao Jing Pan Xinghua(Cell Biological Therapy Center of Kunming General Hospital, Chengdu Military Region of Chinese People＇s Army, Cell Biological Medicine Integrated Engineering Laboratory of State and Region of Yunnan Province, Kunming 650032, China Kunming General Hospital Clinical Collage of Kunming Medical University ,Kunming 650032, China Shanghai Children＇s Hospital, Shanghai 200062, China)

机构地区：[1]成都军区昆明总医院细胞生物治疗中心干细胞与免疫细胞生物医药技术国家地方联合实验室云南省细胞治疗技术转化医学重点实验室,昆明650032 [2]昆明医科大学成都军区昆明总医院临床学院,650032 [3]上海市儿童医院中医科,200062

出　　处：《中华细胞与干细胞杂志（电子版）》2016年第6期369-375,共7页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金(No.81170316)

摘　　要：目的研究人脐带间充质干细胞(hUCMSCs)体外模型中对胶质瘤细胞株U87荷瘤能力及其上皮间质转化(EMT)的影响及其可能机制。方法建立裸鼠皮下荷瘤模型,观察hUCMSCs与U87共培养组和U87单独荷瘤组皮下荷瘤能力的差别,免疫组化检测EMT相关基因组织表达水平,Western Blot检测EMT相关蛋白MMP2,MMP7,MMP9,E-cadherin(E-cad),N-cadherin(N-cad)和Vimentin(VIM)表达情况,q PCR检测EMT相关基因(mmp2,mmp7,mmp9,E-cad,N-cad和vimentin)转录水平。Transwell和Matrigel分别检测hUCMSCs对U87转移能力的影响,两组间比较拟采用独立样本t检验,多组间比较采用单因素方差分析。结果皮下荷瘤结果提示hUCMSCs与U87共培养组皮下荷瘤率高于U87单独荷瘤组,且肿瘤体积较大,RT-PCR结果提示:共培养组和单独培养组相比,E-cad水平下调,N-cad和VIM上调,且基质金属蛋白酶家族MMP2,MMP7,MMP9也有不同程度上调。Western blot结果提示:在蛋白水平共培养组和单独培养组相比,E-cad水平下调2.1倍,N-cad和VIM上调1.0倍,MMP9上调0.9倍;E-cad,N-cad,VIM以及MMP2变化结果与RT-PCR结果一致。免疫组化结果进一步验证hUCMSCs能促进U87表达Ki67,E-cad与N-cad低表达。Transwell和Matrigel实验结果提示hUCMSCs能够促进U87细胞的转移侵袭能力。结论 hUCMSCs能通过促进U87高表达EMT相关基因和增殖相关基因Ki67,从而促进胶质瘤细胞株U87皮下肿瘤形成并发生上皮间质转化,提示hUCMSCs临床应用中应该充分考略受试者是否是潜在胶质瘤患者。Objective To study the effects of human umbilical cord mesenchymal stem cells （hUCMSCs） on growth of human glioma cell line U87 in vitro and epithelial-mesenchymal transition and its possible mechanism. Method The subcutaneous growth of U87 tumor in nude mice was compared between the U87 cells co-cultured with hUCMSCs and U87 cultured alone. The expression of EMT related genes was evaluated with immunohistochemistry. Expression of E-cad, N-cad and MMP9 was evaluated with Western blot for cultrued cells. RT-PCR was used to detect mRNA levels of EMT-associated gene. Using transwell and matrigel methods, the invasion and migration ability of U87 was evaluated. Independent sample t-test and ANOVA were used for comparison of data, which were expressed as mean _ standard deviation. Results IMore subcutaneous tumor was seen in nude mice inoculated with U87 cells co-cultured with hUCMSCs than U87 cells cultured alone, with up- regulated expression of E-cadherin, mmp2, mmp7,and mmp9 and down-regulated expression of N-cad. Western blot showed the same results in E-cadherin,N-cadherin, vimentin and MMP2 changes with RT-PCR. Transwell and Matrigel results showed the migration and invasion of U87 was promoted by hUCMSCs. Conclusion hUCMSCs can promot EMT and proliferation of U87, suggesting that the potential tumor-promoting effects of hUCMSCs should be fully considered in clinical application of hUCMSC.

关 键 词：脐带 间质干细胞 胶质瘤 生物转化 

分 类 号：R739.41[医药卫生—肿瘤]