裸鼠原位肝癌耐药模型的建立

To set up a mdr1 multidrug resistant model of orthotopic transplantation of liver carcinoma in nude mice

目的 建立裸鼠原位肝癌耐药模型。 方法 培养肝癌细胞系HepG2 ,建立裸鼠的皮下肿瘤 ,形成供瘤鼠。开腹直视下将瘤块种植于 2 5只裸鼠的肝包膜下建立原位肝癌模型 ,通过表阿霉素间歇腹腔化疗 ,建立 2 0只裸鼠原位肝癌耐药模型。对照组 5只用体检、B超、CT、剖腹探查监测肝内瘤块生长情况。用逆转录聚合酶链反应和免疫组织化学方法检测肿瘤耐药基因mdr1 mRNA和p gp蛋白的表达。 结果 模型建立无手术死亡 (0 / 2 5 ) ,种植成瘤率为 88% (2 2 / 2 5 ) ,补种 3只全部成功 ,耐药诱导成功率为 80 % (16 / 2 0 )。诱导组 2 0只mdr1 mRNA和p gp蛋白的表达均明显高于对照组 (5只 ) ,分别约是对照组的 2 3倍和 13倍。 结论 成功地建立了与临床肝癌相似的裸鼠原位肝癌耐药模型 ,为进一步研究肝癌多药耐药基因的诊断和逆转提供了良好的动物平台。

Objective To set up a mdr1 multidrug resistant model of orthotopic transplantation of liver carcinoma in nude mice by intermittent abdominal chemotherapy. Methods The hepatocellular carcinoma cell line HepG2 was first cultured, then subcutaneous carcinoma was produced to form the tumor donor mice. An orthotopic mdr1 hepatoma was produced by implanting the tumor fragment subserosally to the mice liver, and intermittent chemotherapy with Pharmorubicin given to induce drug resistance. Physical examination, ultrasonography, spiral CT and laparotomy were used to examine the growth of the tumor. RT-PCR and SP method by the monoclonal antibody JSB-1 were adoped to detect the expression of mdr1-mRNA and p-gp protein. Results There was no mortality. The successful rate of tumor implantation is 88% (22/25), the successful rate of supplementary implantation was 100% (3/3), and the successful rate of induction of drug resistance was 80% (16/20). The expressions of mdr1-mRNA and p-gp in the Pharmorubicin group were 23 folds and 13 folds of the control group, respectively. Conclusions The nude mice mdr1 multidrug resistant model of orthotopic liver carcinoma were set up successfully, with its features similar to clinical cases. It would be helpful for the further study of the diagnosis and reversal strategy of the MDR phenomenon.