乳腺癌干细胞的培养及鉴定  

Culture and identification of breast cancer stem cells

作　　者：郑文博[1] 潘凌霄[1] 唐炜[1] 高进[1] 叶熹罡[1] 

机构地区：[1]广州医学院第一附属医院乳腺外科,广东广州510120

出　　处：《南方医科大学学报》2011年第12期2021-2025,共5页Journal of Southern Medical University

基　　金：广东省医学科学技术研究基金(WSTJJ2006122744010419630407371X)

摘　　要：目的从乳腺癌患者的肿瘤组织中获得乳腺癌干细胞,并确定其生物学特性。方法收集乳腺癌患者的肿瘤组织,利用乳腺球悬浮培养法获得乳腺球细胞。用流式细胞仪检测细胞CD44以及CD24的表达,用Western印迹法检测细胞ALDH1,ESA以及Oct4的表达。将2×104、2×105和2×106三种细胞数的原代乳腺球囊细胞及贴壁细胞分别接种到NOD/SCID鼠的脂肪垫,观察其成瘤能力和肝肺转移情况。结果利用乳腺球悬浮培养的原代乳腺癌细胞中有62.36%呈CD44+/CD24-/low表型,且肿瘤干细胞标记物ALDH1,ESA以及Oct4的表达高于贴壁培养的原代乳腺癌细胞,差异有统计学意义(P<0.05)。与贴壁细胞相比,悬浮培养富集的原代球囊细胞均有更强的成瘤和肝肺转移能力。结论利用乳腺球悬浮培养法可以从人乳腺癌组织中获得乳腺癌干细胞。Objective To isolate breast cancer stem cells from breast cancer patients and identify their biological characteristics.Methods Mammospheric cells were purified and enriched from the tumor tissues of breast cancer patients using mammosphere culture.Their expressions of CD44 and CD24 were analyzed by flow cytometry,and ALDH1,ESA and Oct4 expressions were determined by Western Blotting.The primary mammospheric and adherent cells,at the density of 2×104,2×105 or 2×106,were inoculated into NOD/SCID mice to observe their tumorigenic and metastatic activities.Results With mammosphere culture method,62.36% of the mammospheric cells showed CD44＋/CD24-/low phenotype.The expressions of ALDH1,ESA and Oct4 in the mammospheric cells were significantly higher than those in the adherent culture-derived breast cancer cells（P0.05）.Primary mammospheric cells were at least 100-fold more tumorigenic than the adherent cells;the mammospheric cells were associated with liver or lung metastases,but the adherent cells were not.Conclusions Mammosphere culture can be employed to obtain breast cancer stem cells from the tumor tissues of breast cancer patients.

关 键 词：乳腺癌 肿瘤干细胞 乳腺球悬浮培养 

分 类 号：R737.9[医药卫生—肿瘤]