鼻咽癌CNE1细胞株裸鼠肌肉和皮下移植瘤模型的建立及比较

Significance of Xenograft Tumor Model in the Muscle of Nude Mice to Study the Invasion and Metastasis of Nasopharyngeal Carcinoma Cells

目的建立人类鼻咽癌CNE1细胞系balb/c裸鼠的肌肉移植瘤模型,观察高分化细胞系CNE1在体内的生长及侵袭能力,为下一步研究鼻咽癌亚临床灶奠定基础。方法体外培养LMP1阳性表达的CNE1细胞系,细胞免疫组化法检测LMP1表达情况,将细胞悬液分别注射balb/c裸鼠臀大肌和臀部皮下,各12只。定期观察并测量移植瘤大小,待有裸鼠濒临死亡时处死全部裸鼠取标本,4%多聚甲醛固定后石蜡包埋切片,分别做免疫组化分析及苏木精-伊红染色,观察肿瘤细胞浸润及转移情况。结果与皮下组相比,肌肉组癌细胞向周围侵袭明显,组织切片中可见肌肉组织内癌巢及癌细胞侵入肌细胞现象。肌肉组移植瘤体内平均潜伏期[(11.92±2.02)d]和瘤体平均倍增时间[(2.32±0.24)d]均较皮下组[(15.40±2.46)d、(3.01±0.32)d]明显缩短(均P<0.01);平均瘤重(1.32±0.17)g较皮下组(1.13±0.15)g明显增加(P<0.05);淋巴结和肝脏转移12例,较皮下组(5例)明显增多(P<0.05);成瘤率(12例/10例)二者无显著差异(P>0.05)。结论裸鼠肌肉移植瘤模型对研究鼻咽癌细胞在体内的生长和侵袭能力较为省时、高效。

Objective To establish xenograft tumor model of well-differentiated nasopharyngeal carcinoma cell line （CNE1） in the muscles of babl/c mice in order to investigate the capability of growth and invasion of CNE1 in vivo and lay the foundation for studying the subclinical loci. Methods CNE1 cells expressing LMP1 were cultured in vitro. The expression of LMP1 was detected by using immunocytochemical method. CNE1 cell suspension after amplification in vitro was injected into the gluteus maximus muscle and gluteal hypo. of babl/c nude mice respectively. The size of the xenograft tumors was observed and recorded every day and all the mice were killed once one was dying. The xenograft tumors were taken out, fixed with 4% polyoxymethylene and embedded with paraffin, and cut into sections. The sections were detected by immunohistochemical method and hematoxylin-eosin stain. The invasion and metastasis of tumor cells were analyzed. Results In comparison to the subcutaneous models, the CNE1 cells injected into muscles infiltrated into adjacent tissue obviously, and cancer nest could be seen in the muscle tissue and cancer cellular invasion to muscle cells also could be found in the histological section. The mean latency period [（11.92±2.02） days] and mean doubling time [（2.32±0.24） days] in the xenograft tumors injected into mus- cles were both significantly shortened （P〈0.01）, the average tumor weight increased obviously [（1.32±0.17） g, P〈0. 05], and the metastasis rate of lymph node and liver increased significantly （41.7 %/100%, P〈0. 05）. But there was no significant difference in the tumor formation rate between two groups （83.3 % vs 100%, P〉0.05）. Conclusion The xenograft tumor model in muscle of nude mice was a more economical and effective method to investigate the invasion and metastasis of NPC cells in vivo.