Effect of the low molecular weight heparin combined with cisplatin on the autophagy of liver cancer cells

ObjectiveTo investigate the effects and mechanism of low molecular weight heparin(LMWH) combined with cisplatin(DDP) on the autophagy of human hepatocellular carcinoma(HCC) BEL-7404 cells.

MethodsThe BEL-7404 liver cancer cells were treated with different concentrations of LMWH and DDP alone or in combination for 48 h, and divided into the control group, LMWH group, DDP group and L+D group.CCK-8 method was used to measure the effects on cell proliferation ability.Dansylcadaverine fluorescence staining was used to observe the autophagy levels.The expression of autophagy-related protein mRNA was detected by RT-PCR.Western blotting was used to detect the expression of autophagy-related proteins and pathway proteins.

ResultsCompared with other groups, the cell proliferation ability significantly decreased(P < 0.05), the green fluorescence autophagy body significantly increased, the mRNA and protein expression levels of autophagy-related protein LC3Ⅱ significantly increased(P < 0.01), the p62 significantly decreased(P < 0.01), and the protein expressions of P-AMPK/AMPK, P-ULK1/ULK1 and Beclin1 significantly increased in the L+D group(P < 0.05 to P < 0.01).

ConclusionsThe LMWH combined with DDP can significantly increase the autophagy of BEL-7404 cells, which may be realized through activating AMPK-ULK1-Beclin1 signaling pathway.