一株新的人肾癌原代细胞系的建立及其生物学特性鉴定

Establishment and characterization of a new primary human renal carcinoma cell line

目的:取肾癌病人标本建立一株新的人肾癌细胞系,初步对该细胞系进行鉴定,为进一步的肾癌基础研究提供实验模型。方法:2008-2009年间共采集20例肾癌新鲜手术标本,于手术后1h内将每例标本切取四块0.5cm×0.5cm×0.5cm组织块,分别包埋于2只裸鼠的右后肢皮下及背部皮下,连续传代3次,取移植瘤体外培养,记录细胞株的生长曲线,测定细胞集落形成率,对细胞进行DNA含量测定,进行染色体分析及病理学检查。结果:其中1只裸鼠皮下移植瘤生长,继续传代,肿瘤生长速度明显加快。取移植瘤标本体外培养得到肾癌细胞系XJG-9201。形态结构,分化程度与原发瘤一致,染色体众数为65。细胞群体倍增时间为38.2h,细胞周期分析G1期62.7%,G2期11.2%,S1期25.3%,集落形成率为70%。结论:肾癌细胞系XJG-9201与原发癌保持相同的生物学特性,体外连续传代1年以上传115代。细胞形态不变,生长周期恒定,已成为一个稳定的细胞系。

Objective： To establish a new renal cell carcinoma （RCC）cell line from Specimens of RCC patients and to characterize the biological features in order to build up an experiment model for the further basic research of RCC. Methods： During 2008 to 2009, 20 fresh surgical samples of RCC from patients were obtained; each specimen was cut in four （each tissue 0.5 cm × 0.5 cm × 0.5 cm）, which was inoculated subcutaneously into the right posterior legs and the back of two nude mice within lh after surgery. After three pas- sages in the nude mice, the tumor tissue was cultured in vitro. Record the analysis of cell growth, determine the colony-forming efficiency, make a chromosome analysis and histopathological examination to the line, and/determine DNA content of cell. Results： Among the ;two nude mice, the transplanted tumors in a mouse grew rapidly, continuing to passage. The tissue of the transplanted tumors was cultured in vitro to obtain a stable-growth RCC cell line （XJG-9201）, without any change of morphological structureand and differentiation compared with primary tumour. The chromosome mode was 65; the cell population doubling time was 38.2 hours; the cycle analysis showed G1 62.7%, G2 11.2% and S1 25.3%. and the colony forming efficiency was 70%. Conclusions： XJG-9201 was identical to the primary cancer cell in biological characteristics. It has been cultured in vitro continually for more than one year and passaged for 115 generations .So, XJG-9201 is considered to be a stable cell line.