摘要
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 xenografted model of human hilar cholangiocarcinoma in nude mice. Methods The fresh human hilar cholangiocarcinoma specimen was cultured. The 40th-and 50th-passage cells were subcutaneously transplanted into the nude mice. The transplanted tumors were retransplanted into new mice. Direct dissemination method was used to affirm the karyotype. The ultrastructure was observed under the transmission electron microscope. Carbohydrate antigen 19-9 (CA19-9), carbohydrate antigen 125 (CA125), carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP) were examined in enzyme immunometric assay; vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), proliferating cell nuclear antigen (PCNA), bcl-2 and P53 were determined with immunohistochemistry. Results Only one of the four grafted tumors was successfully passaged in the first passage. The tumor was successfully passaged for 5 times. HE staining showed a poorly differentiated adenocarcinoma structure that just like the human hilar cholangiocarcinoma specimen. The chromosomal analysis revealed an ultratrieuploid human karyotype of 69-81 in number. Ultrastructural analysis showed a typical malignancy characteristic. According to enzyme immunomatric assay, the cellular concentrations of some tumor markers were as follows: CA19-9 > 240 U/ml, CA125 > 500 U/ml, CEA: 54.23 ng/ml, AFP: 13.92 ng/ml. While the four markers were all negatively in the serum. VEGF, bFGF, PCNA, bcl-2 and P53 were strongly expressed. Except for the first passage growth curve, other passages were very similar. Conclusions The xenografted model of hilar cholangiocarcinoma in the nude mice is shown to be useful for further investigation of human hilar cholangiocarcinoma.
出处
《中华肝胆外科杂志》
CAS
CSCD
2005年第6期403-406,共4页
Chinese Journal of Hepatobiliary Surgery
