LY294002抑制P13K／Akt通路对人骨肉瘤类肿瘤干细胞增殖的影响  

LY294002 inhibits proliferation of cancer stem-like cells from human osteosarcoma via down-regulation of PI3K/AKT signaling pathway

作　　者：宫晨[1] 郭风劲[1] 秦亮[1] 祁军[1] 尹德龙[1] 姚斌[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《中华实验外科杂志》2011年第12期2215-2217,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：教育部重点实验室开放基金资助项目（2010KF01）

摘　　要：目的分离鉴定人骨肉瘤类肿瘤干细胞并探讨LY294002对其增殖的作用。方法采用无血清培养法从人骨肉瘤MG-63细胞中分离培养人骨肉瘤类肿瘤干细胞，免疫组织化学检测干细胞表面标志CD44及CD133的表达，CCK-8法观察加入不同浓度磷脂酰肌醇-3-激酶（P13K）抑制剂LY294002（5、10和20umol／L）后对其增殖的影响，并用Westernblot检测总蛋白激酶B（Akt）和磷酸化Akt（p-Akt）的变化。结果无血清培养法可用于分离人骨肉瘤类肿瘤干细胞，人骨肉瘤类肿瘤干细胞CD44及CD133呈阳性表达。LY294002可显著抑制其增殖，细胞生长抑制率平均分别为12．54％、14．92％和19．91％（P〈0．05），作用可能与通过抑制P13K进而抑制Akt的磷酸化有关。结论LY294002对人骨肉瘤类肿瘤干细胞的增殖具有抑制作用。Objective To isolate and identify the cancer stem-like cells （CSCs） from human osteosarcoma, and investigate the impact of LY294002 on their proliferation. Methods CSCs were isolated from human osteosarcoma ceil line MG-63 ceils with serum-free media, and the expression levels of CD44 and CD133 were detected by using immunocytochemistry. The impacts of LY294002 （5, 10 and 20 umol/L）, an inhibitor of PI3K/Akt pathway, on the CSCs proliferation were tested by CCK-8 kit. Western blotting was employed to detect the expression levels of total Akt and p-Akt. Results The human osteosarcoma CSCs could be isolated by serum-free media, and expressed the stem cell surface markers such as CD44 and CD133. LY294002 could inhibit the growth of the human osteosareoma CSCs more significantly than control group, and the inhibition ratio of cell growth was 12. 54%, 14. 92% and 19. 91% respectively （P 〈 0.05）. The inhibition effect might be related to the inhibition of PI3 K and p-Akt. Conclusion LY294002 could inhibit the proliferation of the human osteosarcoma CSCs.

关 键 词：骨肉瘤 肿瘤干细胞 P13K/AKT通路 

分 类 号：R738.1[医药卫生—肿瘤]