干细胞培养基成球培养法筛选结肠癌干细胞的生物学特性  

Isolation and characterization of spheroid cells from the HCT116 colon cancer cell line

作　　者：杨东[1,2] 张金龙[3] 黄轶轩[1] 栾青春[1] 刘健[1] 孟希亭[1] 王海娟[1] 钱海利[1] 李贵新[2] 林晨[1] 

机构地区：[1]中国医学科学院肿瘤医院分子肿瘤学国家重点实验室,北京100021 [2]潍坊医学院 [3]中国人民解放军第302医院

出　　处：《山东医药》2012年第25期12-15,共4页Shandong Medical Journal

基　　金：国家自然科学基金面上项目(81071773;30973471;30973447)

摘　　要：目的探讨干细胞培养基成球培养法筛选结肠癌干细胞的相关蛋白表达及生物学特性。方法用干细胞培养基成球培养法筛选结肠癌干细胞,Western blot检测干细胞相关蛋白的表达,流式细胞仪检测细胞周期;同时检测干细胞增殖、黏附、耐药及侵袭能力,并摸索结肠癌干细胞最佳冻存条件。结果用干细胞培养基成球培养法成功培养出了结肠癌细胞球,检测发现干细胞相关蛋白表达增强,静止期细胞比例明显升高,细胞增殖速度慢,黏附性、耐药性、侵袭性均强于亲本细胞。结论干细胞培养基成球培养法筛选的细胞球中富集了肿瘤干细胞,为结肠癌干细胞的研究提供了良好的细胞模型。Objective To identify the colon cancer stem cells from colon cancer cell lines in SCM and analyzed the stem cell-related biological characteristics. Methods The colon cancer stem cells were isolated and amplified by culturing the clone spheres. Western blot was used to detect the expression of stemness-related proteins and flow eytometry was used to perform cell cycle assay. The capacity of adhesion, drug resistance and invasion of these cells were also evaluated by b^- ological models, at the same time screening the optimal freezing. For colon cancer stem cells. Results Colon cancer clone spheres were successfully amplified and some stem cell-related proteins were expressed at high leve!s. Cells from clone spheres showed a higher portion of cells in phase GO/G1, slower proliferation rate and increased capacity of adhesion, drug resistance and invasion compared with their parental cells. Conclusions The clone sphere cultured in SCM enrich with cancer stem cells and provide a good cell model for colon cancer stem cell research.

关 键 词：结肠癌干细胞 黏附性 耐药性 

分 类 号：R733.3[医药卫生—肿瘤]