CAJ | 학술논문

目的采用无血清培养方法从人结肠癌细胞株中分选出富含肿瘤干细胞的球囊样细胞,并分析其增殖和迁移特性.方法人结肠癌细胞株HCT116、HT29细胞在特殊配制的无血清培养基(SFM)中悬浮培养,观察球囊样细胞的生成.流式细胞仪检测结肠癌干细胞分子标记CD133表达,利用CCK-8比色法以及Transwell小室法研究结肠癌球囊样细胞的增殖情况和迁移能力.采用成组设计资料的t检验分析检测数据.结果 HCT116、HT29细胞在SFM培养条件下可以获得可稳定传代的球囊样细胞,SFM培养下HCT116球囊样细胞CD133阳性细胞占75.44％±11.41％,HT29球囊样细胞CD133阳性细胞占76.22％±14.23％.与常规含血清培养基(SSM)培养的同系细胞比较,HCT116及HT29球囊样细胞中结肠癌干细胞分子标记CD133阳性细胞数明显增多(t=11.43,9.17,P＜0.05),其持续增殖能力和运动迁移能力均增强.结论无血清培养获得的结肠癌球囊样细胞高表达结肠癌干细胞分子标记CD133,且具有更强的增殖和迁移能力.它可作为进一步研究结肠癌干细胞的良好模型.
목적 채용무혈청배양방법종인결장암세포주중분선출부함종류간세포적구낭양세포,병분석기증식화천이특성.방법 인결장암세포주HCT116、HT29세포재특수배제적무혈청배양기(SFM)중현부배양,관찰구낭양세포적생성.류식세포의검측결장암간세포분자표기CD133표체,이용CCK-8비색법이급Transwell소실법연구결장암구낭양세포적증식정황화천이능력.채용성조설계자료적t검험분석검측수거.결과 HCT116、HT29세포재SFM배양조건하가이획득가은정전대적구낭양세포,SFM배양하HCT116구낭양세포CD133양성세포점75.44％±11.41％,HT29구낭양세포CD133양성세포점76.22％±14.23％.여상규함혈청배양기(SSM)배양적동계세포비교,HCT116급HT29구낭양세포중결장암간세포분자표기CD133양성세포수명현증다(t=11.43,9.17,P＜0.05),기지속증식능력화운동천이능력균증강.결론 무혈청배양획득적결장암구낭양세포고표체결장암간세포분자표기CD133,차구유경강적증식화천이능력.타가작위진일보연구결장암간세포적량호모형.
Objective To obtain colon cancer spheroid cells from human colon cancer cell lines cultured in serum-free medium (SFM),and investigate the proliferative and migratory properties of colon cancer spheroid cells.Methods Human colon cancer cell lines HCT116 and HT29 were cultured in SFM,and then the generation of spheroid cells was observed.The expression of stem cell surface marker CD133 was detected by flow cytometry,and the proliferative and migratory properties of colon cancer spheroid cells were detected by cell counting kit-8 and Transwell migration assay,respectively.All data were analyzed by using the t test.Results Spheroid cells were obtained from colon cancer cell lines HCT116 and HT29 in SFM.The ratios of spheroid cells with positive expression of CD133 generated by HCT116 and HT29 were 75.44％ ± 11.41％ and 76.22％ ± 14.23％,respectively.Compared with original colon cancer cells cultured in serum supplemented medium,the number of HCT116 and HT29 spheroid cells with positive expression of CD133 was significantly greater (t =11.43,9.17,P ＜ 0.05 ),and the proliferative and migratory abilities were much stronger also.Conclusion Colon cancer spheroid cells cultured in SFM have higher positive expression of CD133 and stronger proliferative and migratory abilities,and it can be utilized as a feasible model for further studies of colonic stem cells.