Effect Of EGCG On The Expression Of Akt、p-Akt、NF-κB In PI3K/Akt Signal Pathway Of The ACC-M Cell Line

Objective: To detect the cell proliferative activity and the expression of Akt, p-Akt, NF-κB in ACC-M cells, we applied EGCG, PI3 K inhibitor LY294002 and their combination in ACC-M cells, and explored the mechanism of salivary adenoid cystic carcinoma after EGCG treatment. Methods: 1. With different concentrations of EGCG, LY294002 applied in ACC-M cells for 24 h, 48 h, 72 h, MTT was used to determine the cell inhibition rate. Calculated the IC50 of EGCG, LY294002. The experiments were divided into vehicle control group, positive control group(LY294002 IC50 group), EGCG IC50 group, EGCG IC20+LY294002 IC10 group and EGCG IC40+LY294002 IC10 group. 2.The morphological changes of the cells were observed by optical microscope. 3.Flow cytometry was used to detect the apoptosis of the cells in each group. 4.To detect the expression of AKT, p-AKT, NF-κB protein in each group by Western blot. Results: 1. Different concentrations of EGCG(62.5 μm, and 125 μm, and 250 μm, and 500 μm) and LY294002(6.25 μm, and 12.5 μm, and 25 μm, and 50 μm, and 100 μm) role ACC-M cell 24 h, and 48 h, and 72 h respectively, compared to vehicle control group, the inhibition rate of EGCG and LY294002 on ACC-M has statistics differences(P<0.05). The IC50 of EGCG and LY294002 was 156.28 μm and 30.88 μm, respectively. The inhibition rate in 48 h of positive control group, EGCG IC50, EGCG IC20+LY294002 IC10, EGCG IC40+LY294002 IC10 were 51.65%, 51.48%, 31.52% and 56.65%. 2. Cells observed after drug ACC-M cell, the cell number reduction, the cell from change from polygon to fusiform, some apoptosis cells floating in the medium. 3. The apoptosis rate of all drug groups were higher than the vehicle control group(P<0.05); The apoptosis rate of EGCG IC50 was lower than the positive control group(P<0.05) and the apoptosis of EGCG IC40+LY294002 IC10 rate was higher than the positive control group(P<0.05). 4. Compared with the vehicle control group, the expression of p-Akt reduced in all drug groups(P<0.05); The expression of Akt, NF-κB in EGCG IC50, EGCG IC20+LY294002 IC10, EGCG IC40+LY294002 IC10 were lower than the vehicle control group(P<0.05).Conclusion: EGCG, LY294002 can inhibit the proliferation of ACC-M cells, while EGCG IC40+LY294002 IC10 significantly inhibit ACC-M cells growth. The expression of Akt, p-Akt and NF-κB in PI3K/Akt signaling pathway may have some connection with ACC-M cell growth, survival and apoptosis.