Sortilin 1 Regulates HCC Proliferation,migration And Metabolic Reprogramming Through HIF1A Pathway

Background and Purpose of ResearchLiver cancer ranks sixth among the most commonly diagnosed cancers worldwide.The incidence rate of obesity has been increased in recent years,which is also responsible for the increased incidence of HCC.Sortilin 1 is one of the five members of vps10p domain receptor family.Studies have shown that the expression of Sortilin 1 on the surface of hepatocytes can mediate the clearance function of hepatocytes to LDL and reduce the level of plasma cholesterol.The expression of Sortilin 1 on the surface of macrophages promotes macrophage phagocytosis of lipids to form foam cells and participates in the development of atherosclerosis.To explore the function of lipid metabolism related factors in liver cancer can provide molecular basis for clinical analysis.The target gene activated by hypoxia inducible factor HIF1A is involved in many key behaviors of tumor cell biology,including angiogenesis,cell survival,glucose metabolism and invasion.Therefore,we will discuss the expression and function of Sortilin 1 in liver cancer,and elucidate the regulatory effect and possible molecular mechanism of Sortilin 1 on malignant behavior of liver cancer through HIF1A pathway in this study.Method1.To investigate the expression level and prognostic significance of Sortilin 1 in liver cancer.To analyze the gene expression data of TCGA patients and our collected liver cancer tissues,compare the expression level of Sortilin 1 in the adjacent tissues and liver cancer tissues,and determine the correlation between the expression of Sortilin 1 in liver cancer and the survival of patients.The sensitivity and specificity of Sortilin 1 as an early marker of liver cancer were verified by analyzing the expression of Sortilin 1 in adjacent tissues and different stages of liver cancer.The effect of Sortilin 1 expression on the prognosis of patients was detected by survival analysis.2.To explore the effect of Sortilin 1 on the malignant behavior of hepatoma cellsCCK8 and Transwell migration assays were used to detect the effect of Sortilin 1 on the proliferative and migrating ability of hepatoma cells.3.To verify the effect of Sortilin 1 on the metabolism of hepatoma cellsRT-qPCR was used to detect the expression of enzymes and regulatory factors involved in lipid metabolism and glucose metabolism in hepatoma cells after down-regulation of Sortilin 1 expression,and the mRNA level of HIF1A was also detected.In addition,Western blotting was used to detect the protein levels of CPT1A and HMGCS1 to further confirm the regulatory effect of Sortilin 1 knockdown on lipid metabolism.4.To confirm that Sortilin 1 regulates the protein expression level of HIF1AThe efficiency of Sortilin 1 knockout was verified by Western blot and the protein expression level of HIF1A was detected at the same time.The expression of Sortilin 1 in hepatocytes was analyzed by GEO database.Results1.The expression of Sortilin 1 is up-regulated in HCC and indicates the poor prognosis.Firstly,by analyzing the gene expression of TCGA patients with liver cancer,we found that sortilin-1 expression in liver cancer was significantly up-regulated,which was also verified by Western blot.Further analysis showed that sortilin 1 could be used as an early marker of liver cancer and was related to the reduction of total survival time.2.Sortilin 1 regulates the growth rate and migration ability of hepatoma cells.Secondly,we also tested the effect of Sortilin 1 expression on the migration ability of hepatoma cells in vitro.Three different siRNA sequences were used to specifically interfere the expression of Sortilin 1 in SK-Hep 1 cell line,and the cell migration ability was detected by Transwell assay.The results showed that the migration ability of hepatoma cells which interfered with Sortilin 1 expression was inhibited.After transfection,the number of SK-Hep 1 cells that co?Ld migrate between Trans well chambers decreased.The above results suggest that Sortilin 1 has the function of regulating the migration ability of hepatoma cells.3.Sortilin 1 regulates the rate-limiting enzymes of glucose and lipid metabolism.By RT-qPCR and Western blotting,we found that interfering with sortilin 1 can change the activity of lipid and glucose metabolism in hepatoma cells.4.Sortilin 1 regulates HIF1 A.After interfering with the expression of Sortilin 1,RT-qPCR and Western blotting showed that the mRNA and protein levels of HIF1A were significantly decreased.The results of geo analysis showed that the expression of Sortilin 1 was up-regulated with the development of liver pathology.Conlusions:1.Sortilin 1 is upregulated in liver cancer and associated with poor prognosis.2.Sortilin 1 can regulate the proliferation,migration and metabolic activity of liver cancer cells.3.The biological function of Sortilin 1 is related to HIF1A.