摘要
目的:基于磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路探讨肺岩宁方对肺腺癌细胞奥希替尼(AZD9291)耐药的逆转作用。方法:体外培养人肺腺癌细胞系H1975,并建立AZD9291耐药细胞株(H1975/AZD),通过四甲基偶氮唑蓝(MTT)法检测不同浓度肺岩宁方对H1975/AZD细胞活性的影响;将H1975/AZD细胞分为:H1975/AZD组、肺岩宁方低剂量组(10g/L)、肺岩宁方高剂量组(15g/L)、肺岩宁方高剂量+PI3K激活剂(740Y-P)组(15g/L肺岩宁方+30μmoL/L 740Y-P),另取未建立耐药性的细胞作为对照(Control)组。采用CCK8法和克隆形成实验检测各组H1975细胞增殖能力,流式细胞术检测各组H1975细胞凋亡情况,划痕愈合实验检测各组H1975细胞迁移能力,Transwell实验检测各组H1975细胞侵袭情况;qRT-PCR检测各组H1975细胞中Bcl-2、Bax、PI3K、AKT mRNA表达量;Western blot检测各组H1975细胞中Bcl-2、Bax及PI3K/AKT通路相关蛋白表达量。结果:与Control组相比,H1975/AZD组细胞各行为学指标差异均无统计学意义(P>0.05);与H1975/AZD组相比,肺岩宁方低、高剂量组细胞存活率、克隆形成数目、划痕愈合率、侵袭细胞数、Bcl-2 mRNA及蛋白表达量下降,细胞凋亡率、Bax、PI3K、AKT mRNA和Bax、p-PI3K、p-AKT蛋白表达量上升(P<0.05);与肺岩宁方高剂量组相比,肺岩宁方高剂量+740Y-P组细胞中以上各指标趋势得到逆转(P<0.05)。结论:肺岩宁方可通过抑制PI3K/AKT通路激活逆转肺腺癌细胞的奥希替尼耐药性。
Objective:To discuss the reversal effect of Feiyanning formula on the resistance of lung adenocarcinoma cells to osimertinib(AZD9291)based on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway.Methods:Human lung adenocarcinoma cell line H1975 was cultured in vitro,and AZD9291 resistant cell line(H1975/AZD)was established.The effect of different concentrations of Feiyanning formula on the activity of H1975/AZD cells was measured by MTT assay.H1975/AZD cells were grouped into H1975/AZD group,low-dose Feiyanning formula group(10g/L),high-dose Feiyanning formula group(15g/L),high-dose Feiyanning formula+PI3K activator(740Y-P)group(15g/L Feiyanning formula+30μmoL/L 740Y-P),and cells without established drug resistance were marked as the control group.CCK8 method and clone formation assay were performed to detect the proliferation ability of H1975 cells.Flow cytometry was used to measure apoptosis of H1975 cells.The scratch healing experiment was performed to detect the migration ability of H1975 cells in each group.Transwell experiment was performed to measure the invasion of H1975 cells in each group.QRT-PCR was applied to measure the mRNA expression levels of Bcl-2,Bax,PI3K,and AKT in H1975 cells.Western blot was applied to detect Bcl-2,Bax,and PI3K/AKT pathway related proteins in H1975 cells.Results:Compared with the control group,there was no statistically significant difference in various behavioral indicators of cells in the H1975/AZD group(P>0.05).For the H1975/AZD group,the low and high dose Feiyanning formula groups showed a decrease in cell survival rate,clone formation number,scratch healing rate,number of invasive cells,Bcl-2 mRNA and protein,and an increase in cell apoptosis rate,Bax,PI3K,AKT mRNAs,and Bax,p-PI3K,p-AKT proteins(P<0.05).For the high-dose Feiyanning formula group,the trends of the above indicators in the cells of the high-dose Feiyanning formula+740Y-P group were reversed(P<0.05).Conclusion:Feiyanning formula can reverse the resistance of lung adenocarcinoma cells to osimertinib by inhibiting the activation of PI3K/AKT pathway.
作者
白月琴
刘畅
赵艳莉
张梦馨
樊佳鑫
BAI Yueqin(Shaanxi University of Traditional Chinese Medicine Affiliated Hospital Cancer Hospital,Shaanxi Xianyang 712000,China)
出处
《河北医学》
2025年第7期1068-1075,共8页
Hebei Medicine
基金
陕西省教育厅专项科研计划项目,(编号:20JK0600)
陕西中医药大学学科建设基金项目,(编号:2020XK05)。
关键词
肺腺癌
肺岩宁方
奥希替尼
耐药性
磷脂酰肌醇3激酶/蛋白激酶B通路
Lung adenocarcinoma
Feiyanning formula
Osimertinib
Drug resistance
Phosphatidylinositol 3-kinase/protein kinase B pathway
