Objective: To study the inhibitory effect of Hedyotis diffusa on the proliferation of EGFR-TKIs-resistant A549 cells and the regulation of PI3K/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 10 μmol·L-1. The low, middle and high dose groups of Hedyotis diffusa were added with 10, 20 and 30 mL·L-1 of Hedyotis diffusa injection respectively, and the positive group was treated with 10 μmol·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-PI3K, PI3K, 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-PI3K/PI3K 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-PI3K/PI3K and p-Akt/Akt in A549 cells were significantly decreased (P<0.05). Conclusion: Hedyotis diffusa can inhibit the proliferation of EGFR-TKIs-resistant A549 cells, promote apoptosis of A549 cells and induce G0/G1 arrest of A549 cells. The mechanism may related to the inhibition of PI3K/Akt signal pathway.
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