人类黑色素细胞瘤永生化细胞系的建立及生物学特性鉴定

Establishment and Characterization of Human Immortalized Melanocytoma Cell Line

目的建立黑色素细胞瘤细胞系并研究其生物学特性。方法取脑膜黑色素细胞瘤组织原代与传代培养,建立永生化细胞系并检测生物学特性,采用HE染色法、倒置显微镜与透射电子显微镜观察细胞形态及超微结构,采用细胞计数法绘制生长曲线与计算倍增时间,观察细胞染色体核型(染色体显带法)与动物成瘤性,采用SP免疫组织化学法检测Vimentin、S100、HMB45和CK蛋白表达。结果临床病理诊断为颅内原发黑色素细胞瘤,建立的黑色素细胞瘤细胞系呈上皮样细胞生长方式,胞内有黑色素颗粒,可见细胞克隆集落;透射电子显微镜见黑色素小体,倍增时间36h,染色体数量32～110条,细胞系裸鼠皮下移植瘤实验提示未成瘤,且免疫组织化学法染色发现细胞系表达Vimentin为阳性,HMB45、S100、CK为阴性。结论成功建立了黑色素细胞瘤细胞系,有良性肿瘤生物学行为特征。

Objective To establish a human immortalized melanocytoma cell line and explore its biological properties.Methods Meningeal melanoma tissue was isolated to primary and mass culture,to establish a immortalized cell line and to detect its biological characteristics.Cell morphology and ultra-structure were studied through HE staining,inverted microscopy and transmission electron microscopy.Growth curve and doubling time of cells were calculated by cell counting.Karyotype（chromosome banding assay） and tumorigenesis in animals were also detected.Vimentin,S100,HMB45 and CK protein expression were measured by SP immunohistochemical method.Results Clinicopathological diagnosis was primary intracranial melanoma.The melanoma cell line established from melanoma tissue,showed an epithelioid cell growth with intracellular melanin granules,visible cell clone colony and melanosomes;detected by transmission electron microscope.The cell line had a doubling time of 36h,and the chromosome number was 32 to 110.Cell line transplanted into nude mice could not survival and cell line had positive Vimentin expression,and negative HMB45,S100 and CK expression.Conclusion We established a human immortalized melanocytoma cell line with benign biological characteristics.