汉族人肾透明细胞癌细胞系的建立  

Establishment of clear cell renal cell carcinoma lines of Han nationality in China

作　　者：翟羽佳[1] 谭晓洁[1] 侯建国[2] 肖建如[3] 徐丹枫[4] 余永伟[5] 常文军[1] 林丽萍[1] 曹广文[1] 

机构地区：[1]第二军医大学卫生勤务学系流行病学教研室,上海200433 [2]第二军医大学长海医院泌尿外科,上海200433 [3]第二军医大学长征医院骨科,上海200003 [4]第二军医大学长征医院泌尿外科,上海200003 [5]第二军医大学长海医院病理科,上海200433

出　　处：《第二军医大学学报》2008年第5期479-484,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30370788.30571609)~~

摘　　要：目的:取汉族人临床标本,建立肾透明细胞癌(ccRCC)细胞系,初步研究该细胞系的生物学特性。方法:2005～2007年间共采集43例ccRCC新鲜手术标本(包括肾原位肿瘤、骨转移、淋巴结转移肿瘤及癌栓),于手术后30～60 min内用植块法进行体外培养。记录传代超过50代的细胞株的生长曲线,测定集落形成率,对细胞进行染色体分析。病理学检查,流式细胞术检测及NOD-SCID鼠荷瘤实验。结果:绝大部分原代细胞传代次数在5代内.其中5例可连续传代5代以上,4例传代超过10代。仅1例来源于骨转移组织的原代细胞(命名为RCC05-TXJ)及1例来源于原位ccRCC的原代细胞(命名为RCC05- ZYJ)可稳定连续传代。RCC05-TXJ经历21个月传110代,群体倍增时间19.2 h,染色体众数为75,集落形成率为41%;RCC05-ZYJ经历18个月传160代,群体倍增时间为16.5 h,染色体众数为55,集落形成率为37%。免疫组化显示CA9及CD133阳性,流式细胞术分析发现随细胞传代增多CA9及CD133表达明显增高(P<0.05)。两株细胞在NOD-SCID鼠体内均有良好的致瘤及转移性。但是转移倾向明显不同。结论:应用汉族人肾透明细胞癌组织成功建立了2个转移潜能不同的细胞系,随着传代次数增加,肿瘤干细胞比例增加。Objective：To establish clear cell renal cell carcinoma （ccRCC） cell lines from clinical ccRCC specimens of Han nationality in China and to characterize the biological features. Methods： From 2005 to 2007, fresh surgical samples of ccRCC were obtained from 43 patients; the samples included primary tumor in situ, osseous metastasis, lymph node metastasis, and cancerous embolus. The samples were cultured in vitro using explant-culture method within 30-60 min after surgery. Analysis on cell growth and colony-forming efficiency was recorded for the lines which were passaged for over 50 generations. Chromosome examination, pathological examination and tumorigenesis in NOD-SCID mice were used to determine their malignancy. Flow cytometry was used to determine expression of CA9 and CD133. Results： Most of the primary cells could only be passaged for less than 5 gefierations; 5 lines could be serially passaged for over 5 passages, 3 lines for over 10 passages, and only 2 lines could be stably passaged. One line,named RCC05-TXJ, was from osseous metastatic ccRCC and had been serially passaged for 110 generations in 21 months; the average doubling time was 19.2 h,average chromosome number was 75,and colony forming efficiency was 41%. Another line, named RCC05-ZYJ, was from primary ccRCC specimen and had been serially passaged for 160 generations in 18 months; the average doubling time was 16.5 h,average chromosome number was 55, and the colony forming efficiency was 37%. Immunohistological analysis demonstrated that both lines expressed CA9 and CD133. Flow cytometry analysis found that expression levels of CA9 and CD133 increased with the passages. Both RCC05-ZYJ and RCC05-TXJ lines were able to form tumor and to metastasize in NOD-SCID mice; however, their metastatic ability was obviously different. Conclusion： We have established 2 ccRCC cell lines with different metastatic potentials from the clinical ccRCC specimens of Han nationality in China. The ratio of tumor stem cells increases with the passages.

关 键 词：肾透明细胞癌 细胞系 汉族 

分 类 号：R737.11[医药卫生—肿瘤]