The Effect Of Autophagy On TRAIL Induced Hela Cells Apoptosis

Objective:This study is to explore the effect of autophagy on cervical cancer Hela cells apoptosis induced by TNF-related apoptosis inducing ligand( TRAIL), and aimed to offer a new strategy for the clinical treatment for cervical cancer. Methods:? To culture human cervical cancer Hela cells in vitro.? The proliferation activity of Hela cells was detected using MTT assays after treated with different concentrations of TRAIL and TRAIL combined with 3-methyladenine(3-MA) or chloroquine(CQ). The morphologic changes of Hela cells were observed under inverted microscope.? LC3 B and p62 were stained using immunofluorescence(IF) and observed under fluorescence microscope; DAPI staining was used to observe Hela cell apoptosis.? The expression levels of LC3 B, p62 and poly-ADP-ribose polymerse(PARP) were analyzed using Western Blotting.? The data obtained were analyzed using SPSS 17.0 software package, with the measurment data expressed in mean±standard deviation and between-group mean comparisons made with analysis of variance(ANOVA). There was statistical significance if P<0.05. Results:? TRAIL can inhibit the proliferation activity of cervical cancer Hela cells.After different concentrations of TRAIL were applied to Hela cells for 48 hours, it was detected that with the increase of TRAIL concentration, the proliferation activity of Hela cells decreased. IC50 was 6.54 ng/m L, and 2.0 ng/m L(the inhibition rate lower than 50%) was chosen to be follow-up concentration.? The proliferation inhibition effect of TRAIL on Hela cells was potentiated by 3-MA or CQ.It was found that the proliferation activity of Hela cells either in TRAIL+CQ group or in TRAIL+3-MA group was lower than that in single TRAIL or CQ or 3-MA group. There was significant difference with P<0.05.? TRAIL can induce autophagy in Hela cells.The results demonstrated that LC3 aggregations were increased, and the LC3 foci and LC3?protein level were elevated in Hela cells treated with TRAIL, indicating that TRAIL can induce autophagosome formation. Compared with TRAIL group or CQ group, there were noticeable increases in LC3 aggregations, LC3 foci and LC3? protein level in TRAIL+CQ group,which further confirmed that LC3 aggregations were due to autophagy. The decreases of LC3 foci and LC3? protein level in TRAIL+3-MA group and 3-MA group suggested that 3-MA could inhibit autophagy.Compared with control group, the foci and protein level of p62 were significantly decreased in TRAIL group; compared with TRAIL group, they were increased in autophagy inhibitors group. In a word, TRAIL could induce autophagy in Hela cells, while 3-MA as well as CQ could inhibit autophagy.? 3-MA or CQ remarkably enhanced the apoptosis of Hela cells induced by TRAIL.The results of DAPI staining were that the apoptosis rate of Hela cells in TRAIL+3-MA group or TRAIL+CQ group was higher than single TRAIL or 3-MA or CQ group. There was significant difference with P <0.05.? 3-MA or CQ increased the expression of apoptosis related protein induced by TRAIL.The expression of cleavage-PARP in TRAIL+3-MA group and TRAIL+CQ group was higher than in TRAIL group. There was significant difference with P <0.05. Conclusion:1. TRAIL can inhibit the proliferation activity of Hela cells in a dose-dependent way.2. While inducing apoptosis of Hela cells,TRAIL can induce autophagy which plays a protective role in the survival of Hela cells.3. Inhibition of autophagy can enhance the apoptosis of Hela cells induced by TRAIL.