PI3K/Akt/mTOR通路抑制剂对白血病细胞株增殖及其PHLPP表达的影响  

Effect of PI3K/Akt/mTOR signaling pathway inhibitors on proliferation and PHLPP protein expression of leukemia cell lines

作　　者：闫朝奇[1] 张丽君[1] 高峰[1] 潘登[1] 王萍萍[1] 王亚柱[1] 

机构地区：[1]中国医科大学附属第一医院血液科,辽宁沈阳110001

出　　处：《中国肿瘤生物治疗杂志》2012年第4期369-373,共5页Chinese Journal of Cancer Biotherapy

基　　金：辽宁省自然科学基金项目(No.20072105)~~

摘　　要：目的:观察PI3K/Akt/mTOR通路抑制剂wortmannin或rapamycin对白血病细胞株增殖及其PHLPP(PH domainleucine-rich repeat protein phosphatase)蛋白表达的影响。方法:以不同浓度的wortmannin或rapamycin分别作用于人类髓细胞白血病细胞系K562、HL-60,采用WST-1法检测细胞的增殖活性,AnnexinⅤ-FITC双染流式细胞术检测细胞凋亡,Westernblotting法检测细胞中p-Akt、Akt、PHLPP蛋白的表达。结果:Wortmannin以时间以及剂量依赖方式抑制K562、HL-60细胞的增殖(P<0.05),48 h的IC50值分别为(187.6±48.4)、(185.5±48.1)nmol/L。100 nmol/L wortmannin作用于K562细胞、50nmol/L wortmannin作用于HL-60细胞12和24 h后,细胞凋亡率均较对照细胞显著升高[(12.4±0.7)%、(17.6±2.3)%vs(5.0±0.6)%,P<0.05;(11.0±0.2)%、(17.9±1.6)%vs(6.8±0.4)%,P<0.05]。Wortmannin分别作用于K562、HL-60细胞12、24、36 h后,p-Akt、PHLPP的蛋白表达水平明显降低;rapamycin同样可使K562、HL-60细胞中PHLPP蛋白的表达水平降低。结论:PI3K/Akt/mTOR信号通路抑制剂抑制白血病细胞株增殖的同时降低其PHLPP蛋白的表达。Objective：To observe the effect of PI3K/Akt/mTOR pathway inhibitors wortmannin or rapamycin on the proliferation and PHLPP （ PH domain leueine-rich repeat protein phosphatase） protein expression of leukemia cell lines. Methods ： Human leukemia eel1 lines K562 and HL-60 were treated with different concentrations of wortmannin or rapamycin. The proliferation of K562 and HL-60 cells was examined by WST-1 assay and the apoptosis of K562 and HL-60 cells was detected by Annexin V-FITC double staining flow eytometry. The expressions of PHLPP, phosphorate-Akt （p-Akt） and total Akt protein were detected by Western blotting. Results： Wortmannin inhibited the proliferation of K562 and HL- 60 cells in a dose- and time-dependent manner, with the IC50 value of 48 h being （187.6 ±48.4） nmol/L and （185.5 ± 48.1 ） nmol/L （ P 〈 0.05 ）. After treating K562 cells with 100 nmol/L wortmannin and HL-60 with 50 nmol/L wortmannin for 12 and 24 h, the apoptosis rates were significantly higher than those in the control group （ [ 12.4 ± 0.7 ] % , [17.6 ±2.3]% vs [5.0±0.61%,P〈0.05; [11.0±0.2]%, [17.9±1.61% vs [6.8±0.41%,P〈0.05）. After treating K562 and HL-60 cells with wortmannin for 12,24 and 36 h, the protein expressions of p-Akt and PHLPP were significantly reduced. And rapamycin also down-regulated the protein expression of PHLPP in K562 and HL-60 cells. Con- clusion： The inhibitor of PI3 K/Akt/mTOR signaling pathway inhibit the proliferation as well as PHLPP protein expression of leukemia cell lines.

关 键 词：WORTMANNIN RAPAMYCIN 白血病 PI3K Akt mTOR PHLPP 凋亡 

分 类 号：R733.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]