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Exploring the Reversal Effect of Feiyanning Formula on the Resistance of Lung Adenocarcinoma Cells to Osimertinib Based on PI3K/AKT Pathway |
BAI Yueqin, et al |
Shaanxi University of Traditional Chinese Medicine Affiliated Hospital Cancer Hospital, Shaanxi Xianyang 712000, China |
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Abstract 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+30 μmoL/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.
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