人鼻咽癌耐药裸鼠移植瘤模型的建立及耐药性检测

Establishment of drug resistance transplanted tumor models in nude mice with human nasopharyngeal carcinoma CNE-1/DDP and drug resistance detection

目的优化建立人鼻咽癌CNE-1/顺铂耐药裸鼠移植瘤模型的方法,并检测其药耐药性,为后续鼻咽癌的耐药机制研究奠定基础。方法采用顺铂诱导鼻咽癌CNE-1细胞,建立其多药耐药细胞株CNE-1/顺铂细胞;将3个不同浓度的耐药细胞分别接种于裸鼠皮下,观察其成瘤性及转移情况;蛋白印迹法检测LRP、VEGF、Bcl-2在CNE-1细胞、CNE-1/顺铂耐药细胞和移植瘤的表达情况。结果经顺铂诱导建立了鼻咽癌CNE-1/顺铂耐药细胞株;其对顺铂、长春新碱和紫杉醇的耐药性分别增加35.83、23.74和10.54倍。5×10~6/ml的耐药细胞成瘤率为83.33%,其他2个浓度耐药细胞未见成瘤,所有裸鼠均未见肝、肺等器官出现转移瘤。LRP、VEGF和Bcl-2蛋白在耐药细胞和移植瘤中的表达均高于CNE-1细胞（P〈0.05）。结论 CNE-1/顺铂耐药细胞的成瘤性与细胞浓度有关,CNE-1耐药细胞移植瘤依然保持很好的耐药性,可为鼻咽癌耐药逆转剂的研究提供较理想的动物模型。

Objective To look for optimized methods to establish drug resistance transplanted tumor models in nude mice with human nasopharyngeal carcinoma CNE-1 / DDP（ cisplatin）,and to detect its drug resistance in order to set up the foundation for the follow-up researches about the mechanism of drug resistance of nasopharyngeal carcinoma（ NPC）. Methods The multidrug resistant cell line CNE-1 / DDP cells were established by cisplatin induction,then the three different concentrations of drug-resistant cells were inoculated subcutaneously into nude mice, respectively, and to observe the tumorigenicity and metastasis,The expression levels of LRP,VEGF and Bcl-2 in CNE-1 cells,CNE-1 / DDP resistant cells and transplanted tumor were detected by Western Blotting. Results The nasopharyngeal carcinoma CNE-1 / DDP resistant cell lines were successfully established by cisplatin induction,and its drug tolerance to DDP,vincristine and paclitaxel increased35. 83,23. 74 and 10. 54 times,respectively,as compared with that in CNE-1 cells,The successful rate of transplantation tumor was 83. 33% by means of CNE-1 / DDP resistant cells at 5 × 10~6/ ml,however,transplantation tumor was not induced by the other two concentrations of drug-resistant cells,moreover,the tumor metastases in liver,lungs and other organs were not observed in all the nude mice. The expression levels of LRP,VEGF and Bcl-2 in resistant cells and transplanted tumor were significantly higher than those in CNE-1 cells（ P 〈0. 05）. Conclusion The tumorigenicity of CNE-1 / cisplatin resistant cells is related with cells concentration,the transplantation tumor of CNE-1 resistant cells keeps still drug tolerance,which can provide ideal animal models for the researches about reversal agent of drug resistance of nasopharyngeal carcinoma.