人肺腺癌移植瘤多药耐药模型建立及生物学特性的研究

Establishment of multidrug resistant model of human lung adenocarcinoma in nude mice and studies on its biological characteristics

目的:建立耐药特性稳定的肺腺癌移植瘤裸小鼠动物模型,为进行耐药机制研究及逆转药物筛选提供实验模型和实验基础。方法:A549/DDP细胞接种于小鼠右侧腋下,观察肿瘤成瘤情况及生长特性;MTT法检测耐药倍数;RT-PCR和免疫组织化学染色检测小鼠移植瘤多药耐药1(MDR1)和多药耐药相关蛋白1(MRP1)的基因及蛋白表达。结果:移植瘤细胞和原代细胞对DDP的耐药倍数无差异。移植瘤成瘤率均为100%,于第6～9天成瘤(体积>100mm3)。与A549移植瘤相比,A549/DDP移植瘤成瘤潜伏期短(P=0.002),生长速度快。移植瘤中MDR1和MRP1 mRNA及P-糖蛋白(P-gp)和MRP1蛋白的相对表达量均升高,差异有统计学意义,P=0.000。结论:以A549/DDP细胞建立的肺癌多药耐药裸小鼠移植瘤模型,保持耐药活性和基因表型。

OBJECTIVE： To establish a multidrug resistant （MDR） model of human lung adenocarcinoma in nude mice, to provide evidence for studying MDR mechanism and screening of the reversal drugs in vivo. METHODS： MDR human lung adenocarcinoma A549/DDP cells were subcutaneously injected into the right armpit of nude mice. The tumor forming in mice and growth charaetersties of tumors were observed. DDP resistance index （RI） was determined with MTT assay. The expression of MDR1 and MRP1 mRNA and protein were examined respectively by reverse transcriptase polymerase chain reaction （RT-PCR） and immunohistochemistry （IHC）. RESULTS：No difference of RI was found between wild cells and A549/ DDP cell line. Successful tumor forming was achieved in 100% of the animals. Tumors began to form on 6--9 days after transplantation （volume 〉 100 mm3）. Compared with A549 transplanted tumors,the latent period of A549/DDP transplanted tumors were shorter and the rate of growth were faster,indicating significant difference between the two groups （P = 0. 002）. The expression of MDR1, MRP1 mRNA and protein in A549/ DDP transplanted tumors significantly rose （P =0. 000）. CONCLUSION ： The MDR model in nude mice established by A549/DDP ceil line retains the characterstics of MDR phenotype.