Study on the Mechanism of Autophagy of HeLa Cells in Human Cervical Cancer Induced by Fucoxanthin / 中国药房

ABSTRACT

OBJECTIVE:To study the mechanism of autophagy of HeLa cell in human cervical cancer induced by fucoxanthin. METHODS:MTT was adopted to determine the cell activity and calculate the inhibition rate after HeLa cells were cultured by 1, 10,20,40 and 80 μmol/L of fucoxanthin for 48 h. Flow cytometry was used to determine the cell cycle and apoptosis rate after HeLa cells were cultured by 0(blank control),10,20 and 40 μmol/L of fucoxanthin for 48 h;acridine orange staining,LysoTrack-er Red staining,HeLa-GFP-LC3 method and fluorescence microscope were used to observe the autophagy state;Western blot was used to determine the expressions of proteins related to autophagy. RESULTS:The cells had obvious inhibition effect on the cell growth after being cultured by 0,10,20,40 and 80 μmol/L of fucoxanthin. The cell was blocked in G0/G1 stage after being cul-tured by 10,20 and 40μmol/L of fucoxanthin,and had no obvious effect on the apoptosis rate;autophagy degree was increased af-ter the cells were cultured by 40 μmol/L fucoxanthin for 48 h. Compared with blank control,40 μmol/L fucoxanthin could promote LC3Ⅰ transferring into LC3Ⅱ and the expressions of Beclin-1,PTEN,p21;and inhibit the phosphorylation of p-Akt,p-p70S6K and p-mTOR. The pre-treatment by autophagy inhibitor 3-methyladenine(5 mmol/L)could reverse the autophagy of HeLa cells in-duced by fucoxanthin;U0126 could partly reverse the autophagy of HeLa cells induced by fucoxanthin. CONCLUSIONS:Fucoxan-thin can induce the authphagy of HeLa cells by inhibiting Akt signaling pathway and activating MEK/ERK signaling pathway.