灵芝乙醇提取物对A549肺癌细胞的作用及其机制

Effects of Ganoderma lucidum ethanol extracts on A549 cells and its mechanism

目的探讨灵芝乙醇提取物(GLEE)抗肺癌的作用及其机制。方法采用CCK-8法测定GLEE对肺癌A549细胞生长的抑制率;用荧光显微镜观察GLEE对细胞形态的影响;用流式细胞术观察其对细胞周期的影响;用AnnexinV-FITC/PI双染检测细胞的凋亡情况。结果当GLEE浓度为5~320μg·mL^(-1)时,A549细胞生长的抑制率逐步上升,半数抑制浓度(IC_(50))为10.8μg·mL^(-1)。荧光显微镜下观察到不同浓度GLEE作用后的A549细胞出现了细胞皱缩,染色质边缘凝集等凋亡细胞特有的形态学特征。当GLEE的浓度为80、100、200μg·mL^(-1)时,主要将细胞周期阻滞在G_(0)/G_(1)期,当GLEE浓度为300、350μg·mL^(-1)时,主要将细胞周期阻滞在S期。当GLEE浓度为200、300、350μg·mL^(-1)时,早期凋亡率分别为21.66%±5.19%、27.34%±2.89%、28.28%±7.25%,凋亡率显著增加,且呈上升趋势。结论GLEE具有良好的抗肺癌作用,主要作用机制可能为诱导细胞凋亡以及阻滞细胞周期。

OBJECTIVE To explore anti-lung cancer effect and its mechanism of Ganoderma lucidum ethanol extracts(GLEE).METHODS Growth inhibition rate of GLEE on A549 cells was measured by cell counting kit-8.Influence of GLEE on cell morphology was observed by fluorescence microscope.The effect on cell cycle was observed by flow cytometry.AnnexinV-FITC/PI double staining was used to detect cell apoptosis.RESULTS When the concentration of GLEE was 5-320μg·mL^(-1),the growth inhibition rate of A549 cells gradually increased,and the half inhibitory concentration(IC_(50) was 10.8μg·mL^(-1).Under fluorescence microscope,it was observed that A549 cells treated with different concentrations of GLEE showed cell shrinkage,chromatin edge agglutination and other unique morphological characteristics of the apoptotic cells.When the concentration of GLEE was 80,100 and 200μg·mL^(-1),it mainly blocked the cell cycle in G0/G1 phase.When the concentration of GLEE was 300 and 350μg·mL^(-1),it mainly blocked the cell cycle in S phase.When the concentration of GLEE was 200,300 and 350μg·mL^(-1),the early apoptosis rate was 21.66%±5.19%,27.34%±2.89%,and 28.28%±7.25%,and the apoptosis rate increased significantly and showed an upward trend.CONCLUSION GLEE has a good anti-lung cancer effect.The main mechanism of action might be probable to induce cell apoptosis and block cell cycle.