摘要
目的:研究白花蛇舌草对EGFR-TKIs抵抗的A549细胞的增殖抑制作用及对PI3K/Akt通路的调节作用。方法:EGFR-TKIs抵抗的A549细胞分为空白对照组,吉非替尼组,白花蛇舌草低、中、高剂量组及阳性对照组,吉非替尼组加入终浓度为10μmol·L^(-1)的吉非替尼,白花蛇舌草低、中、高剂量组分别加入10、20、30 mL·L^(-1)的白花蛇舌草,阳性对照组加入10μmol·L^(-1)的LY294002,给药48 h后测定各组A549细胞增殖抑制率,Annexin V-碘化丙啶(PI)双染法及脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)染色法测定A549细胞凋亡率,划痕实验测定A549细胞迁移率,流式细胞术测定各组A549细胞周期分布,免疫印迹法(Western blot)测定各组A549细胞p-PI3K、PI3K、p-Akt及Akt水平。结果:与空白对照组比较,吉非替尼组各指标未表现出显著性差异(P>0.05);与空白对照组及吉非替尼组比较,白花蛇舌草各剂量组及阳性对照组A549细胞增殖抑制率、凋亡率及TUNEL阳性A549细胞比例、G0/G1期A549细胞比例显著升高(P<0.05),A549细胞迁移率、S期及G2/M期A549细胞比例、A549细胞p-PI3K/PI3K、p-Akt/Akt水平显著降低(P<0.05);与白花蛇舌草低、中剂量组比较,阳性对照组A549细胞增殖抑制率、凋亡率及TUNEL阳性A549细胞比例、G0/G1期A549细胞比例显著升高(P<0.05),A549细胞迁移率、S期及G2/M期A549细胞比例、A549细胞p-PI3K/PI3K、p-Akt/Akt水平显著降低(P<0.05)。结论:白花蛇舌草能够抑制EGFR-TKIs抵抗的A549细胞增殖,促进A549细胞凋亡,诱导A549细胞G0/G1期阻滞,其机制可能与抑制PI3K/Akt信号通路有关。
Objective:To study the inhibitory effect of Hedyotis diffusa on the proliferation of EGFR-TKIs-resistant A549 cells and the regulation of PI3 K/Akt pathway.Methods:EGFR-TKIs resistant A549 cells were divided into blank control group,low,middle and high dose groups of Hedyotis diffusa and positive group.Gefitinib was added to the gefitinib group at a final concentration of 10μmol·L^(-1).The low,middle and high dose groupsof Hedyotis diffusa were added with 10,20 and 30 mL·L^(-1)of Hedyotis diffusa injection respectively,and the positive group was treated with 10μmol·L^(-1)of LY294002.After incubation for 48 h,the proliferation inhibition rate of A549 cells was determined.The apoptosis rate of A549 cells was determined by Annexin V-propidium iodide(PI)double staining and terminal deoxynucleotidyl transferase mediated Nick-end labeling(TUNEL)staining.Mobility of A549 cell was determination by scratch test.The cell cycle distribution of A549 cells was determined by flow cytometry.The levels of p-PI3 K,PI3 K,p-Akt and Akt in A549 cells were determined by Western blot.Results:Compared with the blank control group,the proliferation inhibition rate,apoptosis rate,the ratio of TUNEL positive A549 cells and the proportion of A549 cells in G0/G1 phase in each dose group and positive control group were significantly increased(P<0.05),while the migration rate of A549 cells,the proportion of A549 cells in S phase and G2/M phase,and the levels of p-PI3 K/PI3 K and p-Akt/Akt in A549 cells were significantly decreased(P<0.05).Compared with the low and middle dose groups of Hedyotis diffusa,the proliferation inhibition rate and apoptosis rate of A549 cells in positive control group,the number of TUNEL positive A549 cells and the proportion of A549 cells in G0/G1 phase were significantly increased(P<0.05),while the migration rate of A549 cells,the proportion of A549 cells in S phase and G2/M phase,and the levels of p-PI3 K/PI3 K and p-Akt/Akt in A549 cells were significantly decreased(P<0.05).Conclusion:Hedyotis diffusa can in
作者
周林水
吕昕
郑苏群
ZHOU Lin-shui;Lü Xin;ZHENG Su-qun(Department of respiratory medicine,the First Affiliated Hospital of Zhejiang University of traditional Chinese Medicine,Hangzhou 310006,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2021年第11期1931-1939,共9页
Chinese Journal of Pharmaceutical Analysis
基金
浙江省自然科学基金项目计划(LQ16H27000)。
