PTEN/Akt/mTOR通路增加K562/ADM细胞化疗敏感性  

PTEN/Akt/mTOR pathway enhances drug sensitivity in K562/ADM cells

作　　者：成志勇[1] 王素云[2] 卞永生[1] 温省初[1] 杨宁[1] 高晓丽[1] 韩英[1] 潘崚[3] 

机构地区：[1]保定市第一医院血液肿瘤科,河北保定071000 [2]河北省人民医院血液内科,河北石家庄050051 [3]华西医科大学华西医院血液内科,四川成都610041

出　　处：《基础医学与临床》2012年第1期49-55,共7页Basic and Clinical Medicine

基　　金：保定市科技攻关计划(11ZF003)

摘　　要：目的探讨K562/ADM细胞系中PTEN/Akt/mTOR信号通路对不同化疗药物敏感性的影响。方法 K562/ADM细胞分为未转染组、转染野生型PTEN组(Ad-PTEN-GFP)、转染空载体组(Ad-GFP),并与不同浓度雷帕霉素或三氧化二砷(As2O3)联合作用。通过MTT法检测细胞增殖,流式细胞术检测细胞凋亡率,荧光定量PCR检测PTEN、mTOR、BCL-2及BAX mRNA水平,Western blot检测PTEN及Akt、p-Akt蛋白水平。结果野生型PTEN对K562/ADM细胞最大增殖抑制率为32.3%,与未转染组及Ad-GFP组相比,Ad-PTEN-GFP组mTOR mRNA和p-Akt蛋白明显减低;雷帕霉素与As2O3联合应用后细胞增殖明显受抑,凋亡率(28.61%±1.46%)明显高于单药作用组(P<0.05),BCL-2 mRNA表达降低,BAX mRNA表达增加,以联合干预组最为明显。结论 PTEN/Akt/mTOR信号传导通路能够增加K562/ADM细胞对雷帕霉素及As2O3的敏感性。Objective To investigate the influence of PTEN/Akt/mTOR pathway in drug sensitivity of K562/ADM cells. Methods K562/ADM cells were divided into untrsfected group, transfected with PTEN group （Ad-PTEN- GFP）, transfected with empty vector group（Ad-GFP）. Each group was treated with rapamycin and or arsenic trioxide at different concentrations. The inhibited rate of K562/ADM cells from each group was measured by MTT assay; the apoptosis rate was assessed by flow cytometry （FCM）. The mRNA expression of PTEN, roTOR, BCL-2 and BAX were detected by real-time fluorescent relative-quantification reverse transcriptional PCR （FQ-PCR）. PTEN, Akt and p-Akt protein levels were detected by western blotting. Results The maximum growth inhibition rate was 32. 3% after transfected with PTEN gene. Compared with Ad-GFP group, after transfacted with Ad-PTEN- GFP into K562/ADM cells, p-Akt protein and roTOR mRNA expression levels were down-regulated. RAPAmycin and arsenic trioxide had synergistic effect on growth inhibition of K562/ADM cells and the inhibition rate（28.61± 1.46% ） was higher than that in cells with single drug treatment （ P 〈 O. 05 ）. BCL-2 mRNA was down-regulated but BAX mRNA up-regulated, especially in combination groups. Conclusions PTEN/Akt/mTOR pathway can en- hance the anti-leukemia effect of rapamycin and arsenic trioxide in K562/ADM cells.

关 键 词：Akt PTEN MTOR 雷帕霉素 三氧化二砷 

分 类 号：R733.7[医药卫生—肿瘤]