食管癌耐药皮下移植瘤裸鼠模型建立

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (1): 77-82.

食管癌耐药皮下移植瘤裸鼠模型建立

刘亮,左静,左连富,郭建文,李金丫

河北医科大学第四医院

收稿日期:2011-04-22 修回日期:2011-08-22 出版日期:2012-01-05 发布日期:2011-12-28
通讯作者: 左连富 E-mail:zuolianfu4909@sina.com
基金资助:
河北省自然科学基金;河北省普通高等学校强势特色学科肿瘤学建设经费

Establishment of esophageal caner drug resistant xenograft nude mice model

Received:2011-04-22 Revised:2011-08-22 Online:2012-01-05 Published:2011-12-28

摘要/Abstract

摘要： [摘要] 目的建立食管癌耐药裸鼠模型及探讨食管癌耐药机制。方法 4周龄的BALB/c nu/nu裸小鼠36只随机分组分为6组，每组6只，左前肢肩胛下皮下分别接种食管癌细胞Eca109及食管癌耐药细胞Eca109/ABCG2，建立裸鼠皮下移植瘤模型。成瘤后腹腔注射阿霉素（Adriamycin, ADM），1、4mg/kg，1次/3d 共注射7次，空白对照组使用生理盐水(normal saline, NS)代替ADM。RT-PCR方法检测移植瘤细胞中三磷酸腺苷结合转运蛋白G2（ATP-binding cassette transporter G2，ABCG2）mRNA 表达情况，流式细胞术(Flow cytometry, FCM)检测移植瘤细胞中ABCG2蛋白、凋亡及细胞中ADM含量。结果成功建立裸鼠食管癌耐药细胞移植瘤模型，皮下接种细胞一周后成瘤，成瘤率100%。实验结束后，注射ADM药物的裸鼠，接种Eca109/ABCG2细胞的皮下移植瘤体积、重量和ABCG2 mRNA、蛋白表达量显著高于接种Eca109细胞移植瘤（P<0.05），但细胞凋亡率及细胞内ADM含量显著降低（P<0.05）。结论接种Eca109/ABCG2细胞的裸鼠皮下移植瘤模型是较好的食管癌耐药动物模型，具有ABCG2耐药表型，为研究ABCG2与食管癌耐药关系提供了较理想的动物模型。

关键词: 食管鳞状细胞癌, 流式细胞术, 多药耐药, 三磷酸腺苷结合转运蛋白G超家族成员2

Abstract: [Abstract] Objective To establish the nude mice model of esophageal cancer drug resistant xenograft and explore the mechanism of esophageal cancer drug resistance. Methods 36 nude mice (BALB/c nu/nu, 4 weeks old) were divided into six groups randomly, 6 each group. Nude mice were respectively inoculated with Eca109 or Eca109/ABCG2 cells subcutaneously in the left subscapularis. xenograft nude mice model was established. The experimental groups received ADM (1, 4mg/kg/3d, respectively), for 7 times. The negative control group received saline. ABCG2 mRNA expression level of xenograft was detected by RT-PCR. ABCG2 protein expression level, cell apoptosis and ADM content in cells of xenograft were detected by flow cytometry. Results The model of esophageal cancer xenograft in nude mice was successfully established, the tumorigenic rate was 100%. After the end of experiment, In the same ADM concentration group, volume, weight and ABCG2 expression of transplanted tumor inoculated with Eca109/ABCG2 cells was significantly higher than transplanted tumor inoculated with Eca109 cells (P<0.05), but the cell apoptosis and ADM content was significantly lower(P<0.05). Conclusion The nude mice xenograft inoculated with Eca109/ABCG2 cells model was an ideal animal model of esophageal cancer drug resistance with ABCG2 resistance phenotype, which provided the ideal animal model for researching the relationship between the ABCG2 and drug resistance of esophageal cancer.

Key words: Esophageal squamous cell carcinoma, Flow cytometry, multidrug resistance, ABCG2

中图分类号:

R735.1

刘亮左静左连富郭建文李金丫. 食管癌耐药皮下移植瘤裸鼠模型建立[J]. 基础医学与临床, 2012, 32(1): 77-82.

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