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目的建立人肝门部胆管癌裸鼠瘤株模型。方法将人肝门部胆管癌标本进行细胞培养,传代。收集第40代、50代瘤细胞接种于4只裸小鼠皮下,并传至5代。各代瘤株行系列检查并与人肝门部胆管癌标本进行比较,观察肿瘤病理结构;采用实体肿瘤体外直接分散法制备染色体;透射电子显微镜观察癌细胞超微结构特点;酶联免疫定量检测法检测糖链抗原19-9(CA19-9)、糖链原125(CA125)、癌胚抗原(CEA)及甲胎蛋白(AFP);免疫组织化学(SABC)法测定血管内皮生长因子(VEGF)、碱性成纤维细胞生长子(bFGF)、增殖细胞核抗原(PCNA)、bcl-2及P53蛋白。结果原代移植成功率为25%(1/4),传代移植成功率为100%。各代肿瘤外观与原代移植瘤相同,呈低分化腺癌表现。染色体主流范围为69～81条。电子显微镜显示符合恶性肿瘤的特点。细胞内CA19-9>240U/ml,CA125>500U/ml,CEA为54.23ng/ml,AFP为13.92ng/ml,血清中此四项均为阴性。VEGF、bFGF、PCNA、bcl-2及P53蛋白表达强度均为“++”。结论第5代肝门部胆管癌裸鼠瘤株可以作为人肝门部胆管癌的动物模型,为胆管癌的基础研究及治疗提供了实验平台。
Objective To establish a model of human hepatocellular cholangiocarcinoma in nude mice. Methods Human hilar cholangiocarcinoma specimens were cultured and passaged. The 40th generation and 50th generation tumor cells were collected and subcutaneously inoculated into 4 nude mice and passaged to passage 5. All the strains of tumor were examined in series and compared with human hilar cholangiocarcinoma specimens to observe the pathological structure of the tumor. Chromosomes were prepared by solid tumor in vitro direct dispersion method. Transmission electron microscopy was used to observe the ultrastructural features of the cancer cells. Enzyme-linked immunosorbent assay (CA19-9), glycogen 125 (CA125), carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP) were detected by enzyme linked immunosorbent assay (ELISA). The expressions of vascular endothelial growth factor ), Basic fibroblast growth factor (bFGF), proliferating cell nuclear antigen (PCNA), bcl-2 and P53 protein. Results The success rate of primary transplantation was 25% (1/4), and the success rate of passage transplantation was 100%. All generations of tumor appearance and the same primary transplanted tumor, showed poorly differentiated adenocarcinoma. The main chromosomal range of 69 to 81. Electron microscopy shows the characteristics of malignant tumors. Intracellular CA19-9> 240U / ml, CA125> 500U / ml, CEA was 54.23ng / ml, AFP was 13.92ng / ml, serum in the four were negative. The expressions of VEGF, bFGF, PCNA, bcl-2 and P53 were all “++”. Conclusion The 5th generation hepatoma cholangiocarcinoma can be used as an animal model of human hilar cholangiocarcinoma and provide an experimental platform for the basic research and treatment of cholangiocarcinoma.