悬浮培养与贴壁培养对乳腺癌MCF-7细胞形态及CD44、CD24表达的影响  

Effects of suspension cultured and adherent cultured breast cancer MCF-7 cell line on cell morphology and cancer stem cell marker CD44,CD24

作　　者：曹露[1] 李传伟[1] 于爱莲[2] 王兆辉[1] 牛丽婷[1] 吴开祥[1] 王学春[1] 

机构地区：[1]山东省泰山医学院病理学教研室,泰安271000 [2]山东省泰山医学院微生物学教研室,泰安271000

出　　处：《临床与实验病理学杂志》2013年第9期969-971,975,共4页Chinese Journal of Clinical and Experimental Pathology

基　　金：山东省科技攻关项目资助课题(2009GG200022047);山东省医药卫生科技发展计划(2011HD015)

摘　　要：目的采用悬浮培养和贴壁培养两种方法分别孵育人乳腺癌MCF-7细胞株,观察细胞形态及干细胞标志物CD44、CD24的变化。方法将乳腺癌MCF-7细胞株同时进行第一代悬浮培养和贴壁培养,用倒置显微镜观察悬浮细胞及贴壁细胞形态及生长方式,收集贴壁细胞及第二代悬浮细胞,对其表面分子CD24及CD44进行免疫荧光染色,荧光显微镜下观察细胞表面标志物的表达,同时采用流式细胞术检测不同亚群的细胞数量。结果悬浮细胞以大小不等的微球方式进行生长,并在第二代悬浮培养中维持此生物学特性;贴壁细胞呈单层贴壁簇状生长。与贴壁细胞相比,CD44分子在悬浮细胞中表达含量增高,CD24分子表达含量减少。CD44+/CD24-/low表型细胞在悬浮细胞和贴壁细胞中所占比例分别为(87.0±0.43)%和(40.1±1.05)%(P<0.05);CD44+/CD24+表型所占比例分别为(11.7±0.39)%和(57.2±0.75)%(P<0.05)。结论悬浮培养法可使乳腺癌MCF-7细胞株贴壁细胞去分化,可富集CD44+/CD24-/low表型的乳腺癌干细胞。Purpose Using suspended culture technology and adherent culture method incubated for MCF-7 human breast cancer cell line, to observed the changes in cell morphology and stem cell molecular marker CD44, CD24. Methods MCF-7 cell line of breast cancer are ordinary cultured and simultaneously suspension cultured to the first generation. The morphology and growth mode of suspen- sion cells and adherent cells were observed using inverted microscope. Adherent cells and second generation of suspended cells were collected and immunofluoreseence stained with its surface antigens CD24 and CD44. The expression of cell surface molecular markers was observed using a fluoreseence microscope and different subsets of cells were detected using flow cytometry at the same time. Re- suits Suspension ceils growed in a way of size microspheres and maintained this biological characteristics in the second generation. Adherent cells were monolayer growed in cluster. Compared with the adherent cells, CIM4 molecule expression level increased in the suspension ceils, and CD24 molecule expression level reduced. The proportion of CD44 ＋/CD24-/low phenotypic cells proportion in sus- pension cells and adherent cells were （ 87.0 -＋ 0. 43 ） % and （40. 1 _＋ 1.05 ） % , respectively （ P 〈 0. 05 ）. Proportion of CD44 ＋ / CD24 ~ phenotype cells were （ 11.7 _＋ 0. 39） % and （57.2 _＋ 0. 75 ） % , respectively （ P 〈 0. 05 ）. Conclusions Suspension culture technology can make the breast cancer MCF-7 adherent cells to dedifferentiate and then enriched CD44 ＋/CD24-/Io,~ breast cancer stem cells.

关 键 词：乳腺肿瘤 癌干细胞 悬浮微球细胞 CD44’ CD24 - low表型细胞 

分 类 号：R737.9[医药卫生—肿瘤]