摘要
目的:检测干细胞相关因子CD44在胃癌细胞株MKN45悬浮球体细胞中的表达。方法:选择人胃癌细胞株MKN45,在含有纤维生长因子-2及表皮生长因子无血清培养液中培养,观察球体形成情况;采用逆转录聚合酶链反应和免疫印迹分析法,检测干细胞相关基因CD44在球体细胞及原代贴壁细胞两组细胞中的表达差异。结果:胃癌细胞株MKN45在无血清培养条件下能形成悬浮球体,球体细胞中CD44的相对表达量高于原代贴壁细胞(P<0.05)。结论:悬浮球培养法在胃癌细胞株MKN45中培养形成的球体细胞富集类肿瘤干细胞。该方法是分选和鉴定肿瘤干细胞的行之有效的方法。
Objective：To detect the expression of stem-cell related factor CD44 in spheroid body-forming cells of gastric cancer cell line MKN45. Methods： Gastric cancer cell line MKN45 were used to culture spheroid bodies in nonad- herent condition in a serum-free medium （SFM）, supplemented with Epidermal Growth Factor （EGF） and Basic Fibroblast Growth Factor（bFGF）. By using RT-PCR and western blot analysis, the expression levels of the embryonic stem cell-related gene CD44 were studied. Results： MKN45 cells formed spheroid bodies in nonadherent condition in a serum-free medium were observed, and the spheroid body-forming cells showed higher expressions of the embryonic stem cell-related gene CD44. Conclusion： Spheroid body-forming cells developed from human gastric cancer cell line MKN45 in nonadherent condition in a serum-free medium supplemented with Epidermal Growth Factor （EGF） and Basic Fibroblast Growth Factor （bFGF） were enriched in cancer stem-like cells. Spheroid body formation assay may be a practical and effective approach to isolate and identify CSCs.
出处
《交通医学》
2013年第6期601-603,606,共4页
Medical Journal of Communications
基金
江苏省南通市科技局资助项目(HS2012067)
