CD133^+结直肠癌SW480细胞系克隆亚系的建立  

Construction of CD133^+ sublines of colorectal cancer cell line SW480

作　　者：李广秋[1] 刘超[1] 郝俊梅[1] 李建明[1] 邓永键[1] 丁彦青[1] 

机构地区：[1]南方医科大学基础医学院病理学系南方医院病理科

出　　处：《诊断病理学杂志》2009年第2期129-133,共5页Chinese Journal of Diagnostic Pathology

基　　金：国家自然科学基金(30670967、30770977);广东省自然科学基金重点项目(5200512);广州市科技计划项目(2007Z3-E4011)

摘　　要：目的建立CD133＋结直肠癌SW480细胞系克隆亚系。方法通过有限稀释法进行单细胞克隆培养，从结直肠癌SW480细胞系48个单细胞孔中培养出8个单细胞源性细胞亚系（编号Y1-a）。对所建立的细胞亚系进行RT-PCR、流式细胞分析及免疫细胞化学筛选CD133＋的细胞亚系（V6、Y2）；并通过MTT法体外增殖实验、划痕实验、体外运动实验和皮下成瘤实验，比较CD133＋之间及与CD133＋单细胞源性细胞亚系的生物学特性。结果筛选出CD133＋单细胞源性细胞亚系2个（Y2、Y6），CD133＋单细胞源性克隆6个（Y1、y3、Y4、K、Y7、Y8）。结论通过单细胞克隆培养建立CD133＋结直肠癌SW480细胞亚系，为下一步结直肠癌转移机制研究提供相关的单细胞源性细胞亚系。Objective To screen CD133 ＋ single-cell derived sublines of eolorectal cancer cell line SW480. Methods 8 single-cell derived sublines （No. Y1-8 ） were screened from 48 single-cell in eolorectal cancer cell line SW480 by limited-dilution method. The CD133＋ cell sublines were detected by RT-CR, flow cytometry, and immtmohistochemistry. Moreover, their biological features were compared with MTT assay, streak experiments, in vitro motility and subcutaneous tumorigenesis. Results There were two single-cell derived sublines （Y2,Y6） were positive for CD133＋ , and the others were negative for CD133. Conclusion CD133 ＋ cell subline of single-cell derived sublines of eolorectal cancer cell line SW480 is constructed by monoclone culture, and it may be importance of understanding of mechanisms for eolorectal cancer metastatasis.

关 键 词：有限稀释法 克隆 亚系 结直肠癌 

分 类 号：R735.35[医药卫生—肿瘤] R734.2[医药卫生—临床医学]