| 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。 |
| 关键词: 肾透明细胞癌 细胞系 汉族 |
| DOI：10.3724/SP.J.1008.2008.00479 |
| 投稿时间：2007-12-29修订日期：2008-03-20 |
| 基金项目:国家自然科学基金（30370788，30571609）. |
|
| Establishment of clear cell renal cell carcinoma lines of Han nationality in China |
| ZHAI Yu-jia1△,TAN Xiao-jie1△,HOU Jian-guo2,XIAO Jian-ru3,XU Dan-feng4,YU Yong-wei5,CHANG Wen-jun1,LIN Li-ping1,CAO Guang-wen1* |
| (1.Department of Epidemiology,Faculty of Medical Services,Second Military Medical University,Shanghai 200433,China;2.Department of Urology,Changhai Hospital,Second Military Medical University,Shanghai 200433;3.Department of Orthopedics,Changzheng Hospital,Second Military Medical University,Shanghai 200003;4.Department of Urology,Changzheng Hospital,Second Military Medical University,Shanghai 200003;5.Department of Pathology,Changhai Hospital,Second Military Medical University,Shanghai 200433) |
| Abstract: |
| 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 generations; 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. |
| Key words: clear cell renal cell carcinoma cell line Han nationality in China |
