人脐带间充质干细胞促进肝癌细胞的增殖和转移  

Human umbilical cord mesenchymal stem cells promote proliferation and metastasis of hepatocelluar carcinoma cells

作　　者：吴宁[1] 杭化莲[2] 张玉玲[3] 戴绘娟 夏强[2] 卞建民[1] 

机构地区：[1]南京医科大学附属南京医院普外科,江苏南京210006 [2]上海交通大学医学院附属仁济医院肝脏外科,上海200127 [3]遵义医学院附属医院贵州省细胞工程重点实验室,贵州遵义563003

出　　处：《南京医科大学学报（自然科学版）》2014年第8期1003-1009,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81100306);上海市科委医学引导类基金(134119a9501);南京市医学科技发展项目(201308014)

摘　　要：目的:研究人脐带来源的间充质干细胞(human umbilical cord mesenchymal stem cells,UC-MSCs)在体内、体外实验中对人肝癌细胞株HepG2的增殖和转移作用。方法:将UC-MSCs在无血清DMEM/F12培养基中培养48 h,收集上清液(MSCsCM)。加入到HepG2的培养基中,使其在HepG2培养基的终浓度为25%、50%和75%作为实验组,DMEM/F12作为对照组,采用CCK-8法检测细胞增殖效应。其后使用Transwell法检测细胞的侵袭效应。建立裸鼠皮下肿瘤模型,健康雌性裸鼠12只随机分为A(HepG2细胞单/左侧皮下成瘤)、B(UC-MSCs+HepG2预混单/左侧皮下成瘤)、C(左侧HepG2、右侧UC-MSCs+HepG2皮下成瘤)3组(n=4),每隔7 d监测成瘤体积变化,50 d后统一处死称量肿瘤湿重。结果:经各浓度MSCs-CM处理后HepG2细胞增殖能力较对照组明显增高(P<0.05);实验组侵袭能力明显高于对照组(P<0.01)。50 d后皮下肿瘤湿重测得A组(0.054 2±0.011 2)g、B组(0.292 0±0.156 9)g、C组[左侧(0.089 4±0.024 2)g、右侧(0.332 6±0.102 9)g],并且从定期体积测量数据发现,两组体积的差异随着时间的推移逐渐增大(P<0.05)。结论:UC-MSCs能促进HepG2的生长和转移,并且这种对肿瘤的增殖和侵袭促进作用可能与UC-MSCs分泌的一些因子有关。Objective：To investigate the effect of human umbilical cord mesenchymal stem cells on the proliferation and metastasis of human hepatocellular carcinoma cell line（HepG2）in vitro and in vivo.Methods：The UC-MSCs were cultured 48 hours in serumfree DMEM / F12 medium,and then the supernatant（MSCs-CM）was collected.The MSCs-CM was added to the culture medium of HepG2 cells to a final concentration of 25%,50% or 75%（the experimental groups）.DMEM / F12 was set as control group.Cell proliferation was detected by cell counting Kit-8（CCK-8）and the migration and invasion were assessed by Transwell assays.Nude mouse subcutaneous tumor models were established.A total of 12 male nude mice were randomly divided into 3 groups（n = 4,respectively）：Group A（subcutaneous injection of HepG2 on the left side）,Group B（subcutaneous injection of UC-MSCs ＋ HepG2 on the left side）and Group C（subcutaneous injection with HepG2 on the left side and UC-MSCs＋HepG2 on the right side）.The volume of tumor was monitored at interval of 7 days until all the mice were sacrificed for the measurements of wet tumor weights at 50 th day.Results：After treatment with each concentration of MSCs-CM,the proliferation of HepG2 was significantly promoted（P〈0.05） and the invasiveness was also significantly improved（P〈0.01）compared with the control group,respectively.Significant differences of wet tumor weight and volume were observed between Group A（0.054 2 ± 0.011 2 g）and Group B（0.292 0 ± 0.156 9 g）,as well as two sides of Group C（left side：0.089 4 ± 0.024 2 g;right side：0.332 6 ± 0.102 9 g,P〈0.05）mice in a time-dependent manner after 50 days.Conclusion：UC-MSCs can significantly promote proliferation and metastasis capability of HepG2 cells via increasing the release of cytokines of UC-MSCs.

关 键 词：脐带间充质干细胞 肝癌 HEPG2 肿瘤增殖 肿瘤转移 

分 类 号：Q257[生物学—细胞生物学]