吉林大学学报(医学版)

• 基础研究 • 上一篇    下一篇

PI3K特异性抑制剂LY294002增强rsTRAIL蛋白对A549细胞的杀伤作用

刘磊1,方艳秋1,齐亚灵2,芦小单1,魏海峰1,谭岩1   

  1. 1.吉林省人民医院肿瘤生物治疗中心,吉林 长春 130021;2.佳木斯大学基础医学院组织学与胚胎学教研室,黑龙江 佳木斯 154007
  • 收稿日期:2014-01-21 出版日期:2014-09-28 发布日期:2014-11-24
  • 通讯作者: 方艳秋(Tel:0431-85595659,E-mail:yq.fang@163.com);齐亚灵(Tel:0454-8618297,E-mail:qiyaling87016@163.com) E-mail:yq.fang@163.com;qiyaling87016@163.com
  • 作者简介:刘 磊(1978-),男,黑龙江省齐齐哈尔市人,医师,医学博士,主要从事肿瘤免疫相关研究。
  • 基金资助:

    吉林省科技厅基础项目资助课题(201115201);吉林省科技厅重点实验室项目资助课题(20122113);吉林省卫生厅科研基金资助课题(20082036);吉林省科技厅科技创新项目资助课题(20082102)

Enhanced effect of rsTRAIL-based therapy for A549 cells by phosphatidylinositol 3′-kinase inhibitor LY294002

LIU Lei1,FANG Yan-qiu1,QI Ya-ling2,LU Xiao-dan1,WEI Hai-feng1,TAN Yan1   

  1. 1.Tumor Biological Treatment Center,Jilin Province People’s Hospital,Changchun 130021,China;2. Department of Histology and Embryology,School of Basic Medical Science,Jiamusi University, Jiamusi 154007,China
  • Received:2014-01-21 Online:2014-09-28 Published:2014-11-24

摘要:

目的:探讨磷脂酰肌醇3激酶(PI3K)参与非小细胞肺癌A549细胞株抵抗重组可溶性肿瘤坏死因子相关凋亡诱导配体(rsTRAIL)蛋白诱导凋亡的机制,寻找增强rsTRAIL蛋白杀伤非小细胞肺癌细胞的新方法。方法:选取对数生长期A549细胞,随机分为rsTRAIL蛋白组和PI3K特异性抑制剂LY294002联合rsTRAIL蛋白(LY294002+rsTRAIL蛋白)组。MTT法检测2组A549细胞生长抑制率,流式细胞技术检测2组A549细胞周期和细胞凋亡的变化,蛋白质印迹法检测2组A549细胞中p-Akt(Ser473)、c-FLIPL、Bcl-2和Bax蛋白表达水平。结果:与rsTRAIL蛋白组(5.16%±0.32%)比较,LY294002+rsTRAIL蛋白组A549细胞生长抑制率(74.6%±2.63%)明显升高(P<0.05)。与rsTRAIL蛋白组比较,LY294002+rsTRAIL蛋白组处理2 h后G0/G1期A549细胞百分比增加(P<0.05),S期细胞百分比降低(P<0.05)。LY294002+rsTRAIL蛋白组A549细胞凋亡率为(61.50±3.02)%,显著高于rsTRAIL蛋白组(3.21%±0.96%)(P<0.05)。蛋白质印迹检测,与rsTRAIL蛋白组比较,LY294002+rsTRAIL蛋白组A549细胞中p-Akt、c-FLIPL和Bcl-2蛋白表达水平降低(P<0.05),Bax/Bcl-2比值升高(P<0.05)。结论:LY294002能够抑制PI3K活性,增强rsTRAIL蛋白对A549细胞的杀伤作用。

关键词: 癌,非小细胞肺, A549细胞株, 肿瘤坏死因子相关凋亡诱导配体, 磷脂酰肌醇3激酶, LY294002

Abstract:

Abstract:Objective To explore the potential mechanisms of non-small cell lung carcinoma cells to rsTRAIL protein-induced apoptosis by phosphatidylinositol 3′-kinase (PI3K/Akt) inhibitor LY294002, and to provide new ways to increase killing activities of rsTRAIL protein for non-small cell lung cancer.Methods The A549 cells at logarithmic growth phase were selected and randomly divided into rsTRAIL group and LY294002+rsTRAIL group.The inhibitory rate of  growth of the A549 cells was tested by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry analysis.The expression levels of Ser473 phosphorylated form of Akt(p-Akt),c-FLIPL protein and Bcl-2 protein in the A549 cells in two groups were analyzed by Western blotting method.Results The inhibitory rate of  growth of the A549 cells in LY294002+rsTRAI group (74.6%±2.63%)was higher than that in rsTRAIL group (5.61%±0.32%) (P<0.05).Compared with rsTRAIL group,the percentage of the cells at G0/G1 phase  in LY294002+rsTRAIL group was increased(P<0.05) and the percentage  of the cells at S phase was decreased(P<0.05).The apoptotic rate of the A549 cells in LY294002+rsTRAIL group (61.5%±3.02%) was higher than that in rsTRAIL group (3.21%±0.96%) (P<0.05). The Western blotting results showed that the expression levels of p-Akt,c-FLIPL  and Bcl-2 proteins in the A549 cells in LY294002+rsTRAIL group were decreased (P<0.05) and the  ratio of Bax/Bcl-2 was increased (P<0.05) compared with rsTRAIL group.Conclusion LY294002 can increase the killing activity of rsTRAIL protein in A549 cells by inhibiting the activity of PI3K.

Key words: cancer non- small cell lung;A549 cells;TNF-related apoptosis inducing ligand; LY294002;PI3K/Akt ,  

中图分类号: 

  • R733.6