磁标记骨髓间充质干细胞对肝细胞癌的抑制作用

Magnetic Labeling Bone Mesenchymal Stem Cells Exert Potent Antitumorigenic Effects in a Model of Hepatocellular Carcinoma

目的应用超顺磁性氧化铁粒子(SPIO)标记骨髓间充质干细胞(BMSCs)移植肝癌大鼠模型,探讨BM-SCs对肝癌细胞的抑制作用。材料与方法培养大鼠BMSCs,菲立磁(Feridex)标记。制备大鼠肝癌模型30只,随机平均分为3组:实验组经脾植入Feridex标记的BMSCs;磷酸盐缓冲液(PBS)组移植等量的PBS;对照组不作任何处理。各组大鼠在BMSCs移植前和移植后1周及2周行MRI,计算各时点肿瘤体积,并记录其存活期。不同时间段大鼠灌注取材,分别做HE染色、普鲁士蓝染色和BMSCs表面标记检测(CD44、CD90)。结果BMSCs的磁标记率近100%,大鼠肝癌模型成功制备。实验组BMSCs移植后1周及2周的肿瘤平均体积明显小于PBS组及对照组(P<0.05)。各组大鼠平均生存时间为:实验组(34.3±5.4)天;PBS组(25.7±3.9)天,对照组(23.5±3.1)天。普鲁士蓝染色显示实验组肿瘤边缘有大量蓝染颗粒,CD44、CD90呈阳性表达。结论BMSCs对肝癌细胞的生长有抑制作用,能够延长肝癌大鼠模型的生存期。

Objective To label rat bone marrow mesenchymal stem cells with superparamagnetic iron oxide（SPIO） and to explore the curative effects of BMSCs for hepatocellular carcinoma cells after transplantation in vivo.Materials and Methods BMSCs from bone marrow of Sprague-Dawly（SD） rats were cultured isolated and purified.Labeled BMSCs was achieved with SPIO（Feridex）.The 30 hepatocellular carcinoma models of SD rats were induced two weeks before transplantation.The models were divided into 3 groups in random：the labeled BMSCs and PBS were transplanted respectively into the rat＇s livers of experimental group and PBS group via spleens,and nothing to do for control group.MR imaging was performed before and 1,2 weeks after transplantation at 1.5T MR system.Volume of tumor on MR imaging obtained before and after transplantation were measured and compared,and survival time of the rats also were compared.After MR imaging,a rat of each group was killed and performed prussian blue staining and detection of the surface markers（CD44、CD90）.Resu-lts Prussian blue staining confirmed the labeling efficiency was above 90%.The proliferation speed of tumor cells in experimental group was slower than that of PBS group and control group（P0.05） in 1,2 weeks after transplantation.Survival time was（34.3±5.4）days in experimental group,（25.7±3.9）days in PBS group,and（23.5±3.1）days in control group.A larger number of prussian blue staining positive cells were found in hepatocellular carcinoma in experimental group,and immunohistochemistry showed positive expression of CD44 and CD90.Conclusion BMSCs have definite therapeutic effect on hepatocellular carcinoma in rats,and also prolong the survival time of model of hepatocellular Carcinoma.