骨髓间充质干细胞与小鼠肝癌肝移植术后复发关系的实验观察

Relationship between mesenchymal stem cells and liver cancer recurrence after liver transplantation in mice

目的 探讨骨髓间充质干细胞（MSCs）与小鼠肝癌肝移植术后复发的关系.方法 利用肿瘤细胞爪垫移植成瘤后切除的方法建立肝癌肝移植术后复发模型.建模成功后将绿色萤光蛋白（GFP）标记MSCs注入615小鼠骨髓腔内,体视荧光显微镜观察MSCs在骨髓腔内增殖情况,肿瘤切除时经小鼠尾静脉注入经转化红色荧光蛋白（RFP）标记的615小鼠MSCs细胞,体视荧光显微镜定期观察肝脏复发前后GFP、RFP标记的MSCs在体内移行情况,复发后,处死小鼠,提取局部复发灶,病理切片证实复发灶病理类型.结果 利用肿瘤细胞爪垫移植成瘤后切除的方法可成功建立肝癌肝移植术后复发模型,造模成功率为37.5%,复发小鼠模型的肝脏大体标本和病理切片检查均符合肝癌表现.对30只有肝癌复发的小鼠通过GFP和RFP双通道荧光检测,4例检测出两种荧光活性、1例检测出GFP活性、25例未检测出任何荧光活性.结论 实验结果发现经骨髓腔内与尾静脉途径进入体内的MSCs可参与肝癌肝移植术后的复发;提示肿瘤宿主体内MSCs是肝癌肝移植术后复发可能的重要来源之一,可能是目前传统＂残存肿瘤细胞＂复发转移理论的完善与补充.

Objective To explore the relationship between mesenchymal stem cells （MSCs） and liver cancer recurrence after liver transplantation in mice. Methods The recurrent murine model of hepatocellular carcinoma （HCC） after liver transplantation was established by transplanting tumor cells of hind paw pads. MSCs labeled with green fluorescent protein （GFP） were injected into the marrow cavity of 615 mice after successful modeling. And the proliferation of MSCs in marrow cavity was observed under stereoscopic fluorescence microscope. MSCs labeled with red fluorescent protein （RFP） were injected into tail vein of mice during tumor dissection. The migration of GFP and RFP- labeled MSCs were tracked before and after tumor recurrence. After recurrence, the mice were sacrificed and the recurrent lesions harvested for conforming pathological type by biopsy. Results The rate of success modeling was 37. 5%. Both gross morphology and pathological examination corresponded to typical HCC manifestations. Thirty mice were detected by GFP/RFP fluorescence for a recurrence of HCC. The outcomes were GFP ＋ RFP （n = 4 ）, GFP （n = 1 ） and neither （ n = 25 ）. Conclusions The presence of MSCs in host may be one of important reasons for recurrent HCC after liver transplantation. It helps to support the traditional view of residual tumor cells mediating the relapse and metastasis of HCC.