Wistar大鼠自发性乳腺肿瘤细胞系的建立及其生物学特性  

Estabishment of a Cell Line ZHL2006 from a Spontaneous Breast Tumor in Wistar Rat and Its Characterization

作　　者：杨文祥[1] 唐利军[1] 赵磊[2] 张金明[1] 孙凡中[1] 吴芳洁[1] 易平[1] 

机构地区：[1]湖北省疾病预防控制中心,武汉430079 [2]华中农业大学,武汉430070

出　　处：《中国实验动物学报》2008年第5期334-337,I0003,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：湖北省自然科学基金(编号:2001ABB133)

摘　　要：目的从Wistar大鼠自发性乳腺肿瘤中分离出乳腺肿瘤细胞系(ZHL2006),并对其进行鉴定,为动物模型的开发、肿瘤发病机制的探讨提供有价值的线索。方法对一例Wistar大鼠自发性乳腺腺癌瘤组织进行培养,对传代培养细胞进行细胞生长曲线测定、细胞形态结构分析、细胞染色体分析,检测广谱细胞角蛋白(PCK)表达、软琼脂集落形成实验等检测。将细胞接种于10只裸鼠,并对形成的肿瘤进行病理组织学检查,并进行分离培养。结果ZHL2006细胞在体外生长迅速,群体倍增时间为43.6 h;倒置显微镜及Giemsa染色镜下观察细胞为多边形及梭形;透射电镜观察可见细胞表面有微绒毛,核高度不规则,核浆比例增大,核仁明显等;染色体数目和结构异常,具有癌细胞特性。广谱细胞角蛋白(PCK)染色阳性,说明其自上皮而非来自基质;同时ZHL2006细胞系在软琼脂中可形成克隆,具裸鼠致瘤性。该乳腺肿瘤细胞系经体外长期培养后已形成永生化细胞系,并具有明显的恶性细胞表型。裸鼠接种实验成功率为6/10,复制肿瘤细胞和原发肿瘤形态结构一致,细胞培养形态结构一致。结论本研究成功的建立了大鼠乳腺肿瘤细胞系,该细胞系具有典型的恶性肿瘤特征,能应用于裸鼠并成功复制出乳腺肿瘤模型。Objective To establish a cell line from a spontaneous breast tumor in a rat, and to characterize this cell line. Methods Tumor cells were isolated form a spontaneous breast tumor in a Wistar rat and cultured, and the growth curve, morphology of the cultured cells, chromosome abnormalities, expression of PCK, and colony formation in soft agar were determined. The established cell line was named ZHL2006. ZHL2006 cells were inoculated into nude mice, and the formed tumor was examined by histopathology. Results The ZHL2006 cells grew fast in vitro and the population doubling time was 43.6 h. Microvilli on cell surface, remarkably irregular nuclei, prominent nucleoli, and high nucleocytoplasmic ratio were seen under the transmission electron microscope, and chromosome number and structure were abnormal with malignant cell characteristics. Positive PCK staining showed that the tumor cells were originated from epithelial cells but not matrix. The ZHL2006 cells could form colony in soft agar and had tumorigenicity in nude mice. After long-term in vitro cultivation the ceils became immortalized with conspicuous malignant cell phenotype. The success rate of tumor formation after inoculation in nude mice was 6/10, and the morphology of tumor ceils were similar to that of original tumor. Conclusion A breast tumor cell line ZHL2006 has been successfully established, showing typical characteristics of malignancy and can be inoculated and to form transplanted tumors in nude mice.

关 键 词：WISTAR大鼠 自发性乳腺肿瘤 细胞系 

分 类 号：R737.9[医药卫生—肿瘤]