迷迭香酸甲酯通过抑制Akt/mTOR信号通路 诱导人肝癌细胞凋亡

Objective To investigate the cell death-inducing effect of methyl rosmarinate (MR) on human hepatoma Hep-3B and SK-Hepl cells and their potential mechanisms. Methods The effects of MR on the viability of Hep-3B, SK-Hepl and MIHA cells were determined by cell counting kit-8 (CCK-8) assay. The morphological changes of three kinds of cells treated with different concentrations of MR were observed by optical microscopy. EdU assay and flow cytometry were used to detect the proliferation and apoptosis of Hep-3B and SK-Hepl cells. Transwell assay was used to study the effects of MR on the migration and invasion of Hep-3B and SK-Hepl cells. Western blotting was used to evaluate the protein expression levels of apoptosis, EMT and Akt/mTOR signaling pathways. Results After treated with different concentrations of MR (0 - 200 Fmol/L) for 48 h, Hep-3B and SK-Hepl cells activities were significantly decreased in a concentration-dependent manner (P<0.01) , while there was no significant effect on MIHA cell activity (P>0.05) , and the IC5„ of Hep-3B and SK-Hepl cells were 102.5 and 99. 3 Fmol/L, respectively. MR treatment (0 - 150 Fmol/L) significantly inhibited the proliferation of Hep-3B and SK-Hepl cells (P<0.05) , while cell detachment and shrinkage were observed by optical microscopy on the Hep-3B and SK-Hepl cells, while the morphology of MIHA cells was not changed. Compared with the control group, MRG00, 150 Fmol/L) induced apoptosis in Hep-3B and SK-Hepl cells, the expression levels of the pro-apoptotic proteins Bax and cleaved PARP were significantly increased (P<0.05) , while the expression level of the anti-apoptotic protein Bcl-2 was significantly decreased (P<0.05). MRG00, 150 Fmol/L) also inhibited the migration and invasion of HCC cells, significantly increased the expression of E-cadherin and decreased the expression of N-cadherin and Vimentin compared with the control group (P<0.05) . Finally, Western blotting results showed that the expressions of p-Akt and p-mTOR in Hep-3B and SK-Hepl treated by MR were significantly reduced in a dose-dependent manner, suggesting that MR may play an anti-cancer role by inhibiting Akt/mTOR signaling pathway. Conclusion MR can promote apoptosis in Hep-3B and SK-Hepl cells, which may be closely related to the inhibition of Akt/ mTOR signaling pathway.