Melittin对非小细胞肺癌增殖、凋亡及PI3K/Akt信号通路的影响  

Effect of melittin on the proliferation,apoptosis and PI3K/Akt signal pathway in non-small cell lung cancer

作　　者：李爱明[1] 赵惠民[1] 揭俊卿[1] 

机构地区：[1]河北保定保定市第一医院胸外科,071000

出　　处：《临床肿瘤学杂志》2015年第4期307-311,共5页Chinese Clinical Oncology

摘　　要：目的探讨Melittin对非小细胞肺癌（NSCLC）细胞增殖、凋亡及PI3K/Akt信号通路的影响。方法分别采用0、10、20、50、100μmol/L Melittin处理NSCLC细胞株A549、SPC-A1及人肺上皮细胞株16HBE 24、48、72和96 h后,采用四甲基偶氮唑盐（MTT）比色法检测各细胞株的增殖抑制率变化,同时采用流式细胞术Annexin-FITC/PI双染法及PI单染法检测不同浓度Melittin处理24、48 h后的A549细胞凋亡及细胞周期情况,Western blotting检测不同浓度Melittin处理48 h后的A549细胞中PI3K/Akt信号通路相关蛋白（Akt和PTEN）及凋亡促进基因（caspase-9）的表达情况。结果在10~100μmol/L范围内,Melittin可呈剂量和时间依赖的方式提高A549、SPC-A1细胞的增殖抑制率,差异均有统计学意义（P〈0.05）,但对16HBE无细胞毒性作用（P〉0.05）;与0μmol/L比较,除10μmol/L Melittin处理24 h后的晚期凋亡率和G2/M期细胞比例无统计学差异（P〉0.05）,10~100μmol/L的早、晚期凋亡率、G0/G1期细胞比例及PTEN和caspase-9蛋白水平均升高,S期、G2/M期细胞比例及Akt水平均降低,以上差异有统计学意义（P〈0.05）,且10~100μmol/L范围内各浓度间的差异均有统计学意义（P〈0.05）。结论 Melittin可对NSCLC细胞有毒性作用,但对正常肺上皮细胞无影响,且可诱导A549细胞凋亡及细胞周期阻滞并抑制PI3K/Akt信号通路的激活。Objective To investigate the effect of melittin on proliferation,apoptosis and PI3K/Akt signal pathway in nonsmall cell lung cancer（ NSCLC）. Methods The NSCLC cell lines A549,SPC-A1 and human lung epithelial cell line 16 HBE were treated with different doses of melittin（0,10,20,50,100 μmol / L）. The Methyl thiazolyl tetrazolium（ MTT） was used to determine the changes of proliferation inhibition rate at 24,48,72 and 96 h after the treatment of melittin. Meanwhile,the Annexin-FITC / PI double staining method and PI single staining method were used to measure the apoptotic rates and cell cycle via flow cytometry. Western blotting was used to detect the protein levels of Akt,PTEN and caspase-9 at 48 h after treatment with different concentrations of melittin. Results Melittin elevated the proliferation inhibition rates in a dose-and time-dependent manner on A549 and SPC-A1 cells（P 〈 0. 05） without cytotoxic effect on 16HBE（P 〉 0. 05）. In addition to similar proportion of G2/ M phase and late apoptotic rate at24 h of 10 μmol / L,there were increased early and late apoptosis rates,cell percentage of G0/ G1 phase and protein levels of PTEN and caspase-9 but decreased cell percentage of S and G2/ M phases and Akt level in 10-100 μmol / L melittin versus 0 μmol / L melittin with statistical significant differences（ P 〈 0. 05）. Significant difference was observed on the above indices during the concentration of 10-100 μmol / L（ P 〈 0. 05）. Conclusion Melittin have toxic effects on the NSCLC cells,but have no effect on normal cells. It can induce the apoptosis and cell cycle arrest of A549 cell with the effects of inhibiting the PI3 K / Akt signaling pathway.

关 键 词：MELITTIN 非小细胞肺癌 增殖 凋亡 PI3K/AKT信号通路 

分 类 号：R734.2[医药卫生—肿瘤]