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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。
Objective To study the inhibitory effect of myricetin on the proliferation of human lung cancer cell line H460 and its possible mechanism. Methods Human lung adenocarcinoma H460 cells were used as in vitro study. MTT assay was used to study the inhibitory effect of different concentrations of myricetin on the growth of H460 cells and the IC50 values were determined. On the phosphorylation of phosphorylated Akt, ERK and the expression of cyclinD1 protein, and the cell cycle was detected by flow cytometry. Results Myricetin significantly inhibited the growth of H460 cells. With the increase of myricetin concentration, the cell growth inhibition rate was significantly increased. The IC50 of myricetin for 72 h was 60 μg / ml. Flow cytometry showed that myricetin significantly reduced the proportion of H460 cells in G2 / M phase and significantly increased the proportion of G0 / G1 and S phases. H460 cells could not enter G2 / M phase under high dose (60 and 90 μg / ml) myricetin treatment . Western blot analysis confirmed that myricetin significantly inhibited ERK phosphorylation, down-regulation of Cyclin D1 expression levels. Conclusion Myricetin inhibits the proliferation of H460 cells in a dose-dependent manner by inhibiting cell entry into G2 / M phase. The ERK-Cyclin D1 pathway may play an important role in the cell cycle regulation of myricetin.