抑制Akt磷酸化对肿瘤坏死因子相关凋亡诱导配体诱导表皮生长因子受体/KRAS野生型非小细胞肺癌H1299细胞凋亡的影响  

Effect of inhibition of Akt phosphorylation on apoptosis of non-small cell lung cancer H1299 cells with wild type EGFR and KRAS induced by tumor necrosis factor related apoptosis inducing ligand

作　　者：邓立力[1] 邓洪滨[1] 韩红霞[1] 李逸文[2] 郭扬[1] 

机构地区：[1]哈尔滨医科大学附属第二医院肿瘤内科,150086 [2]哈尔滨医科大学附属第一医院科教科,150001

出　　处：《肿瘤研究与临床》2016年第3期154-157,共4页Cancer Research and Clinic

基　　金：Science and Technology Research Project of Commission of Education of Heilongjiang Province，黑龙江省教育厅科学技术研究项目

摘　　要：目的 探讨抑制Akt磷酸化对肿瘤坏死因子相关凋亡诱导配体（TRAIL）引发的表皮生长因子受体（EGFR）/KRAS野生型非小细胞肺癌（NSCLC） H1299细胞凋亡的影响.方法 采用Annexin V-FITC/碘化丙啶（PI）法分析TRAIL对H1299细胞凋亡的作用;Western blot检测TRAIL作用后细胞Akt、p-Akt蛋白的表达.使用PI3K特异性抑制剂LY294002处理H1299细胞,用Annexin V-FITC/PI法分析其对TRAIL诱导细胞凋亡的影响;Western blot检测细胞Akt、p-Akt蛋白表达的变化.结果 H1299细胞对TRAIL引起的凋亡呈现耐受.当TRAIL质量浓度为100 ng/ ml时,实验组细胞凋亡率高于对照组[（15.06±1.29）％比（3.56±0.50）％,t=66.953,P=0.000].当TRAIL质量浓度增加到500 ng/ ml时,与100 ng/ ml TRAIL处理组相比,差异无统计学意义[（18.65±2.09）％比（15.06±1.29）％,t=2.423,P=0.136].TRAIL以时间依赖的方式上调H1299细胞的p-Akt表达水平.用LY294002预处理细胞后,TRAIL激发的Akt磷酸化被抑制到基线水平,同时细胞凋亡率明显高于仅TRAIL处理组[（41.65±4.62）％比（15.82±0.61）％,t=39.028,P=0.001].结论 TRAIL引发的Akt磷酸化激活可抵抗TRAIL引起的凋亡反应.抑制Akt磷酸化可明显增强EGFR/KRAS野生型NSCLC细胞对TRAIL引起的凋亡反应的敏感性.Objective To explore the effect of inhibiting Akt phosphorylation on tumor necrosis factor related apoptosis inducing ligand （TRAIL）-induced apoptosis of human non-small cell lung cancer （NSCLC） H1299 cells with wild type EGFR and KRAS.Methods The TRAIL-induced apoptosis was examined by the Annexin V-FITC/PI.The expressions of TRAIL-activated Akt phosphorylation and p-Akt were measured by Western blot.After cells were treated with LY294002, an inhibitor of PI3K-Akt pathway, Annexin V-FITC/PI and Western blot were used to analyze the alteration of TRAIL-induced apoptosis and Akt phosphorylation, respectively.Results H1299 cells were not sensitive to TRAIL-induced apoptosis.When TRAIL concentration was 100 ng/ml, the apoptosis rate of the test group was significantly higher than that of the control group [（15.06±1.29） % vs （3.56±0.50） %, t =66.953, P =0.000].When TRAIL concentration was 500 ng/ml, the difference was not statistically significant compared with apoptosis rate of 100 ng/ml TRAIL group [（18.65±2.09） % vs （15.06±1.29） %, t =2.423, P =0.136].The expression level of Akt phosphorylation in H1299 cells was increased by TRAIL in a time-dependent way.When cells were pretreated with LY294002, TRAIL-induced Akt phosphorylation was suppressed to baseline level.At the same time, the apoptosis rate in LY294002-treated group was significantly higher than that in TRAIL group [（41.65±4.62） % vs （15.82±0.61） %, t =39.028, P =0.001].Conclusions TRAIL-induced Akt phosphorylation can antagonize TRAIL-induced apoptosis.Inhibition of Akt phosphorylation can significantly enhance the sensitivity of NSCLC H1299 cells with wild type EGFR and KRAS to TRAIL-induced apoptosis.

关 键 词：癌 非小细胞肺 AKT 肿瘤坏死因子相关凋亡诱导配体 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]