结直肠肿瘤细胞球的干细胞相关特性研究  

Biological characteristics of human colorectal cancer spheres cultured in serum-free medium

作　　者：贾健锋[1] 程勇[1] 郑梦颖[1] 饶明月[1] 

机构地区：[1]重庆医科大学附属第一医院胃肠外科,重庆400016

出　　处：《重庆医科大学学报》2011年第11期1339-1342,共4页Journal of Chongqing Medical University

基　　金：重庆市自然科学基金资助项目(编号:2008BB5230)

摘　　要：目的:无血清悬浮培养生成5种人结直肠肿瘤细胞球,体外稳定传代培养,研究其在体外增殖、分化及转移等生物学特性。方法:人结直肠癌细胞SW480、SW620、LOVO、HT29和HCT116在添加了细胞生长因子的无血清培养基(Serum-free medi-um,SFM)中悬浮培养,稳定传代并诱导分化。选择其中HCT116细胞系用流式细胞技术检测单层细胞及其肿瘤细胞球干细胞标记分子CD133+的表达。Transwell小室实验、免疫荧光及RT-PCR分别检测HCT116单层细胞及其肿瘤细胞球的体外转移能力,以及间质化标记分子N-cadherin及Vimentin的表达。结果:5种人结直肠癌细胞系均在SFM中形成能稳定传代培养的肿瘤细胞球,其在含血清培养基(Serum supplemented medium,SSM)中重新贴壁生长并各自恢复亲代细胞的生物学特性。与HCT116单层细胞相比,HCT116肿瘤细胞球中CD133+细胞比例显著增多,并具有更强的转移能力,以及高表达间质化标记分子N-cad-herin及Vimentin。结论:通过无血清培养可以生成人结直肠肿瘤细胞球,这些球体中富含肿瘤干细胞样特性的细胞。Objective：To generate cancer spheres from five different colorectal cancer cell lines in serum-free medium（SFM）,stably subculture them in vitro,and study their biological characteristics of proliferation,differentiation and metastasis.Methods：Human colorectal cells SW480,SW620,LOVO,HT29 and HCT116 were cultured in SFM with supplemented cell growth factors;stable subculture was induced to differentiate in serum supplemented medium（SSM）.The expression of CD133＋ in HCT116 monolayer cells and spheres were investigated by flow cytometry.Cell metastasis assay,immunofluorescence and RT-PCR were applied to examine cell metastasis ability in vitro and the expression of mesenchymal markers N-cadherin and Vimentin of both HCT116 monolayer cells and spheres.Results：Cancer spheres could be generated from five colorectal cancer cell lines in SFM.These spheres were stably subcultured and induced to differentiate by serum supplemented medium,and these differentiated cells were in adherent growth and kept the same morphology with the parental cells.HCT116 spheres,compared with parental cells,had higher CD133＋,N-cadherin and Vimentin expressions as well as metastasis ability.Conclusion：Colorectal cancer spheres can be generated in SFM culture,and these cancer spheres are rich in cancer stem-like cells.

关 键 词：结直肠肿瘤 肿瘤干细胞 上皮-间质化 无血清培养 CD133 

分 类 号：R735.35[医药卫生—肿瘤] R735.31[医药卫生—临床医学]