肝癌姑息性切除术后裸鼠残癌模型的建立

目的 建立肝癌姑息性切除术(PR)后可研究残癌生物学特性变化的转移性裸鼠人肝癌原位移植瘤模型.方法 采用单肝叶双瘤源原位接种技术建立MHCC97H原位移植瘤模型,PR切除1瘤,观察5周.测量肿瘤大小,流式细胞术计数外周血循环肿瘤细胞数,荧光显微镜和苏木素-0伊红(HE)染色检测肝内、肺、腹腔转移等,免疫组织化学染色和Western blot检测脑和肿瘤组织Vimentin表达.结果 双瘤源接种成瘤率95.6％、PR成功率95.3％、裸鼠5周总体存活率90.2％.假手术组、肝部分切除术组、PR组肿瘤大小、循环肿瘤细胞数均依次升高,肝内转移率分别为15.4％、41.7％、83.3％,肺转移率分别为61.5％、91.7％、100.0％,腹腔转移率分别为23.1％、50.0％、83.3％(P值均＜0.05).手术组脑和肿瘤组织Vimentin表达均显著上调.结论 成功建立转移性裸鼠人肝癌原位移植瘤合并PR模型,可用于研究残癌生物学特性的变化.

Establishment of metastatic human hepatocellular carcinoma orthotopic transplantation model with palliative resection for investigating the biological characteristics of residual cancer in nude mice

Objective To construct a metastatic human hepatocellular carcinoma (HCC) orthotopic transplantation model with palliative resection (PR) in nude mice for investigating the biological characteristics of the residual cancer,and to verify its efficacy.Methods A metastatic human HCC orthotopic nude mice model (MHCC97H) was established by using the method of bi-tumor inoculation in a single liver lobe.Two weeks after transplantation,the PR was applied to resect one tumor from the two.lobes.The tumor size was measured five weeks postresection.The flow cytometry was used to count the circulating tumor cells (CTCs) in peripheral blood.The fluorescence microscope and hematoxylin and eosin (HE)staining were adopted to determine the metastasis of the liver,lung,or abdomen.The Vimentin expression in brain or tumor tissues was detected by using immunohistochemical staining or Western blotting respectively.Results The incidence of bi-tumor formation,successful PR,and 5-week overall survival were 95.6％,95.3％,and 90.2％ respectively.The tumor size and CTCs in sham operation group,liver partial hepatectomy group,and PR group were increased in gradient; the incidence of intrahepatic metastasis was 15.4％,41.7％ and 83.3％ ; lung metastasis rate was 61.5％,91.7％ and 100.0％ ; and abdomen metastasis rate was 23.1％,50.0％ and 83.3％,respectively (P＜0.05).The Vimentin expression in brain or tumor tissues was up-regulated after surgical resection.Conclusion A metastatic human HCC orthotopic nude mice model combined with PR is successfully constructed,and it may be used to investigate the biological characteristics of the residual cancer.