Cholesterol Attenuated HCC Progression Through Modulating IP3Rs Mediated Calcium Release

Background: Worldwide,hepatocellular carcinoma(HCC)remains a top instigator of cancer mortality.Previous clinical studies have revealed that low serum cholesterol predicts a poor outcome in HCC patients,but the potential role of cholesterol in the progression of HCC remains controversial.Methods: In vivo,we applied HCD and atorvastatin to treated HCC mice induced by DEN.In vitro,SK-Hep1 and Huh7 cells were treated with LDL and atorvastatin.The proliferation ability was tested with CCK-8 assay.The migration ability was examined with transwell and wound healing assay.Furthermore,a comprehensive analysis of transcriptome sequence and metabolomics was adapted to explore the mechanisms that cholesterol inhibits HCC progression.JC-1 staining was applied to tested mitochondria membrane potential.Flou-4 AM and mitotracker co-staining was adapted to tested calcium concentration in mitochondria.Results: In vivo,HCD inhibited HCC progression which was reversed by atorvastatin(Quantification of tumors: NCD 6.25±5.04/liver;HCD 2.11±1.37/liver;HCD+atorvastatin 6.78±5.61;HFD+atorvastatin 27.00±18.77,n=8,p<0.05).In vitro,LDL inhibited migration and proliferation of HCC cell lines which could also be reversed by atorvastatin(SK-Hep1,LDL at 6h and 12 h are 12.86±1.60% and 42.2±2.95% respectively,CTL at 6h and 12 h are 18.69±1.14% and 49.27±1.79%).Furthermore,the comprehension analysis of transcriptome sequencing and metabolomics revealed that IP3 Rs participated in the inhibition of cholesterol on HCC progression.Following the line,cholesterol inhibited mitochondrial calcium concentration and mitochondrial function(SK-Hep1,CTL vs LDL,3.40±0.34 vs 2.66±0.31 p<0.05,n=5;Huh7,CTL vs LDL 2.83±0.24 vs 1.85±0.31,p<0.05,n=5).And then LDL inhibited mitochondrial calcium concentration and function were reversed by ATP,an agonist on IP3 Rs mediated calcium release(SK-Hep1,CTL vs LDL,3.40±0.34 vs 2.66±0.31 p<0.05,n=5;Huh7,CTL vs LDL,2.83±0.24 vs 1.85±0.31,p<0.05,n=5).Conclusion: Cholesterol inhibited HCC progression through modulating IP3 Rs meditating calcium release which was essential to maintain normal mitochondrial function.