LY294002对人纤维肉瘤HT1080细胞生长的影响及机制研究  

Effect and Mechanism of LY294002 on Growth of Fibrosarcoma Cell Line HT1080

作　　者：周虹[1,2] 黄向阳[3] 杨婷[1,2] 汪涛[1,2] 徐丹[1,2] 文富强[1,2] 

机构地区：[1]四川大学华西医院呼吸内科,成都610041 [2]四川大学华西医院生物治疗国家重点实验室呼吸病学研究室,成都610041 [3]四川大学华西医院风湿免疫科,成都610041

出　　处：《四川大学学报（医学版）》2010年第1期20-23,共4页Journal of Sichuan University(Medical Sciences)

基　　金：教育部博士点基金资助

摘　　要：目的研究PI3K特异性抑制剂LY294002对人纤维肉瘤HT1080细胞生长的影响及机制。方法将对数生长期的HT1080细胞分组培养,对照组加入不含LY294002的培养液,实验组加入LY294002,浓度分别为5、10、25、50、100μmol/L,培养12 h和24 h后,用SunBioTMAm-Blue法测定细胞增殖抑制率。100μmol/LLY294002作用HT1080细胞24 h后,观察细胞形态的改变,用流式细胞仪检测细胞凋亡情况,并收集蛋白标本,Western Blotting检测p-Akt及p-mTOR蛋白的表达。结果与对照组比较,随着LY294002剂量、作用时间的增加,对HT1080细胞增殖抑制率增大(P<0.05)。LY294002(100μmol/L)作用24 h后HT1080细胞胞体皱缩,细胞数较对照组减少,细胞早期凋亡率较对照组增加(P<0.05),p-Akt(0.23±0.01)及p-mTOR(0.32±0.06)蛋白的表达低于对照组p-Akt(0.63±0.02)及p-mTOR(0.71±0.02)蛋白的表达(P<0.01)。结论LY294002通过抑制PI3K-mTOR信号通路来抑制HT1080细胞的生长,PI3K-mTOR信号通路参与纤维肉瘤细胞的生长调节,该通路可能作为治疗纤维肉瘤的新靶点。Objective To investigate the effect and mechanism of LY294002 on growth of fibrosarcoma cell line HT1080. Methods The proliferation inhibitory rate of HT1080 cells treated by LY294002 at doses of 5,10, 25,50,100 μmol/L for 12 and 24 h, respectively, was evaluated using SunBioTM Am-Blue method. HT1080 cells were divided into two groups, that is, A and B. The A group was control group without treatment. The B group received LY294002 （100 μmol/L） for 24 h. The changes of cell morphology and quantity were observed by phase contrast microscope. The apoptosis rate of HT1080 cells was detected by flow eytometry. And the protein expression of p-Akt and p-mTOR in HT1080 cells were detected by Western Blotting. Results The proliferation of HT1080 cells was inhibited in time- and dose- dependent manner by LY294002. After the treatment of 100 ptmol/L LY294002 for 24 h, the growth of HT1080 cell line was remarkably inhibited by LY294002. The rate of apoptosis increased. And the protein expression of p-Akt （0.2±0. 01） and p-mTOR （0.32±0. 06） in LY294002 group was lower than p-Akt （0.63±0.02） and p-mTOR （0.71±0.02） in control group （P〈0.01）. Conclusion LY294002 can inhibit the growth of HT1080 cells through PI3K-mTOR pathway. PI3K-mTOR pathway presents an appealing therapeutic target on fibrosarcoma.

关 键 词：LY294002 纤维肉瘤 HT1080 PI3K-mTOR 

分 类 号：R73-3[医药卫生—肿瘤]