杨梅素抑制H460人肺腺癌细胞增殖及细胞外信号调节激酶通路作用

目的研究杨梅素(myricetin)对人肺癌H460细胞的增殖抑制作用、及其作用途径。方法以人肺腺癌系H460细胞为离体研究对象,通过噻唑蓝(MTT)法研究不同浓度的杨梅素对H460细胞生长的抑制作用并测定其半数抑制浓度(IC50);Western blot法检测杨梅素对蛋白激酶B(Akt)、细胞外信号调节激酶(ERK)磷酸化水平及cyclinD1蛋白表达的影响;流式细胞术检测细胞周期。结果杨梅素对H460细胞具有明显的抑制作用,随着杨梅素浓度的增加,细胞的生长抑制率明显升高,杨梅素作用72h的IC50值为60μg/ml。流式检测发现,杨梅素可明显降低G2/M期H460细胞比例,明显增加G0/G1和S期比例,高剂量(60、90μg/ml)杨梅素处理下,H460细胞无法进入G2/M期。蛋白印迹检测证实,杨梅素干预明显抑制ERK磷酸化、下调Cyclin D1表达水平。结论杨梅素剂量依赖性抑制H460细胞增殖与其抑制细胞进入G2/M期有关,ERK-Cyclin D1途径可能在杨梅素细胞周期调控中发挥重要作用。

INHIBITION OF MYRICETIN ON HUMAN LUNG ADENOCARCINOMA H460 CELL PROLIFERATION AND THE ROLE OF ERK PATHWAY

Objective To investigate the effects of myricetin on the H460 lung adenocarcinoma cell proliferation and clarify the underlying mechanism.Method The inhibitory effects of myricetin on the growth of H460 cells were investigated and its value of IC50 after 72 h incubation were measured by monotetrazolium(MTT) method.The levels of pAkt (phosphorylated protein kinase B)Ser473,PERK (phosphorylated extracellular signal-regulated kinase)1/2,cyclin D1,total Akt and ERK1/2 proteins were detected by Western blotting.The cell cycle was investigated by flow cytometry.Results Myricetin inbibited the H460 cell proliferation in a dose-dependent way.The IC50 of myricetin treatment for 72 h was 60μg/ml.Myricetin treatment remarkably reduced the percentage of cells in G2/M phase.It was found that myricetin remarkbly reduced the phosphorylation of ERK1/2,and cyclin D1 protein level,but did not influence the level of phospholated AktSer473.Conclusion Myricetin could inhibit the growth of H460 cell in a dose-dependent way,inhibite the G2/M progression.ERK-Cyclin D1 pathway may play an important role in regulation of cell cycle in H460 by myricein.