LY294002联合AD-PTEN对脑胶质瘤细胞增殖和侵袭的影响及机制探讨  

Inhibition effect and mechanism of LY294002 and Ad-PTEN on proliferation and invasion activity in glioma cell line TJ889 cell

作　　者：宋云朋[1] 钟跃[1] 刘哲[1] 潘强[1] 

机构地区：[1]天津医科大学总医院,天津300052

出　　处：《山东医药》2010年第2期1-3,共3页Shandong Medical Journal

基　　金：天津市科委应用基础计划资助项目(09JCZDJC17600)

摘　　要：目的观察PI3K抑制剂LY294002联合含PTEN基因的重组腺病毒(Ad-PTEN)对人脑胶质瘤TJ899细胞增殖和侵袭的影响,并探讨其机制。方法向TJ889分别加入LY294002(LY294002组)、Ad-PTEN+LY294002(联合组)、Ad-vector病毒(空载组)、DMSO(DMSO组)培养48 h。分别采用MTT法、流式细胞术、划痕实验和Tran-swell小室观察TJ899增殖活性和侵袭能力的变化;Western Blot检测TJ899的PTEN/PI3K/AKT信号转导通路中相关蛋白。结果与DMSO组、空载组和LY294002组相比,联合组细胞存活速率下降,细胞周期阻滞在G0/G1期,S期减少,细胞凋亡率增加,侵袭和迁移的细胞数减少,PTEN蛋白表达增加,而磷酸化蛋白激酶B、细胞增殖抗原、细胞周期素D1、Bcl-2、基质金属蛋白酶-2、黏着斑激酶蛋白表达显著下降,P均<0.05。结论LY294002联合Ad-PTEN可明显抑制TJ899的增殖活性和侵袭能力,该作用与抑制PI3K/AKT信号通路有关。Objective To observe the inhibition effeet and mechanism of LY294002 and Ad-PTEN on proliferation and invasion in glioma cell line TJ889 cell. Methods Ad-PTEN were transfected into TJ889, being treated by LY294002. The proliferation and invasion activity of TJ889 cells was examined by MTI＇, Flow Cytometry , Annexin V methods, Transwell and scarification test respectively. Western blot analysis was used to detect the protein expression in PI3K/AKT signal pathway. Results The cell multiplication rate in combination group presented decreasing trend since the sixth day in culture （P 〈 0.05 ）, as compared with DMSO, Ad-vector and LY294002 group. The number of cells inhibited at G0/G1 phase in combination group was more than any other group （ P 〈 0.05）. The apoptosis rate in combination group was higher than that in the other three groups （ P 〈 0.05）. Transwell and Scarification test indicated that the invasion and migration ability of the cells in combination group were decreased obviously （P 〈 0.05）. The protein expression of PTEN in combination group were significantly higher, while PCNA,Cyclin D1 ,bcl-2 and MMP9,p-FAK were lower than those in the other three groups （ P 〈 0.05 ）. Conclusion Combine application of LY294002 and Ad-PTEN can enhance the effect of inhibition to glioma, it may be related to the inhibitory to the PI3K/Akt pathway.

关 键 词：颅内肿瘤 胶质瘤 酪氨酸磷酸酶基因 PI3K抑制剂 LY294002 信号通路 

分 类 号：R739.41[医药卫生—肿瘤]