建立双标记乳头状甲状腺癌裸鼠转移模型的初步探索  

Exploration of establishing a metastatic nude mouse model of double-tagging papillary thyroid cancer

作　　者：罗丹丹 苏贞辉[2] 赵丽芳 高芸 王海[1] 管庆波[1] 于春晓[1] 

机构地区：[1]山东大学附属省立医院内分泌科,山东省内分泌与脂代谢重点实验室,山东省临床医学研究,济南250021 [2]山东大学附属省立医院病理科 [3]辽宁何氏医学院生物化学系

出　　处：《中华内分泌代谢杂志》2017年第11期970-975,共6页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金青年项目（81101590）;国家自然科学基金面上项目（81641030、81770860）

摘　　要：目的建立稳定整合生物发光和红色荧光的乳头状甲状腺癌细胞株，并构建适用于小动物活体成像系统观察的裸鼠乳头状甲状腺癌转移瘤模型。方法利用慢病毒将携带有荧光素酶基因和红色荧光蛋白基因的重组质粒感染恶性程度较高的乳头状甲状腺癌细胞株BHP10．3SCmice，潮霉素筛选出稳定高表达荧光素酶和红色荧光蛋白的细胞。接种于裸鼠大腿肌肉，建立乳头状甲状腺癌转移瘤模型，通过小动物活体成像进行在体生物发光成像和离体荧光成像．系统监测肿瘤的生长及转移过程。结果获得了高水平稳定表达荧光素酶和红色荧光蛋白的乳头状甲状腺癌细胞BHP10—3mluc，该细胞与BHP10—3SCmice细胞具有相似的生长特性。将BHP10—3mluc细胞接种于裸鼠肌肉内可以成瘤并转移至周围淋巴结及肺脏，小动物活体成像系统能准确监测肿瘤细胞在体内的生长及转移过程。结论成功获得稳定表达生物发光和红色荧光的乳头状甲状腺癌细胞，并建立了适用于小动物活体成像系统观察的裸鼠乳头状甲状腺癌移植瘤转移模型，为活体动态评估乳头状甲状腺癌的生长、转移及药物疗效提供了理想模型。ObjectiveTo obtain a thyroid cancer cell line integrated with luciferase and fluorescence, and to establish a metastatic nude mouse model of papillary thyroid cancer monitored with in vivo imaging system.MethodsLentivirus carrying recombinant plasmid containing luciferase gene and mCherry gene infected high malignant papillary thyroid cancer cell line （BHP10-3SCmice）. The stable thyroid cancer cell line （BHP10-3mluc）labeled with luciferase and mCherry selected by hygromycin, analyzed for fluorescence with fluorescence microscopy and bioluminescence with in vitro analysis and in vivo imaging. 0.25 ml（5×10^6/ml）BHP10-3mluc cell suspension were injected into the left thigh intramuscular tissue of 5-week-old nude mice to establish BALB/C nude mice metastatic tumor model. The growth and metastasis of the tumors were monitored weekly with in vivo imaging system. The nude mice were sacrificed on the 42th day and the tissues with metastatic tumors were analyzed by fluorescent imaging ex vivo. The ectopic implanted tumors and metastatic lesions were verified by tissue sections with Hematoxylin and Eosin staining.ResultsBHP10-3mluc cells showed similar growth characteristics to the original BHP10-3SCmice cells and stably expressed luciferase and mCherry. At the end of the first week after ectopic implantation, the xenograft tumors were found and in the 6th week, the tumors were found to metastasized to adjacent lymph nodes and lungs, which was consistent with the results of pathology.The growth and metastasis of tumors can be accurately monitored with in vivo imaging system.ConclusionsA PTC cell line stably expressing bioluciferase and fluorescence was successfully established.The nude mouse model of PTC metastatic tumor generated by intramuscular inoculation of those cells can be well monitored with in vivo imaging system, which provides ideal models for researches on tumor growth, metastasis, and drug treatment.

关 键 词：乳头状甲状腺癌 移植瘤 在体生物发光成像 离体荧光成像 

分 类 号：R-332[医药卫生] R736.1