人鼻咽癌耐药裸鼠移植瘤模型的建立及耐药性检测

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。5×106／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。

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

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 increased 35.83,23.74 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 ×106/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 .