The Metabolomics Analysis And The Migration Mechanism Exploration Of Hepatocellular Carcinoma Cells While Administrated By Arginine Deprivation

Liver cancer is the sixth most common cancer in the world and the second most common cause of death.More than 700,000 people a year have died of liver cancer in recent years,with more than half of all new cases and deaths occurring in China.Traditional treatment methods,such as surgical resection and chemotherapy,are likely to cause cancer metastasis and drug resistance and less effective for patients with advanced hepatoma cell.Thus,finding new treatment methods for HCC are increasingly urgent.Recent years,with the development of targeted gene therapy for cancer,arginine deprivation by specific drugs have showed a promising prospect to cure the cancer of arginine auxotroph,which has entered the phase III clinical trials in HCC,however,the resistance to HCC have find frequently,which influenced the effects of arginine deprivation in clinical trials.First,non-targeted LC-MS metabolomics was used to detecte the normal hepatic cell L02,hepatocellular carcinoma cells Bel7402 and Hep G2,which had administrated by arginine deprivation.Then,standardized treatment followed by multivariate statistical analysis continued.Next,the P<0.05 and VIP≥1 were used as the standard to screen the significantly differential ions and then the metabolites were identified.By using Metaboanlyst database,the metabolites used for analyzing the effects of arginine deprivation on HCC,which led to the study of arginine deprivation promoting the stress response pathway in HCC.Western blot and RT-PCR performed to verify the effect of arginine deprivation on p38α key factor and the expression of its downstream ITGB8,and then the effect of arginine deprivation on the migration of HCC testified by wound healing and cell counting experiments.The aim of this study was to investigate metabolomics analysis and the migration mechanism exploration of HCC while administrated by arginine deprivation.Multivariate statistical analysis indicated that arginine deprivation had little effect on the metabolism of L02 and had a great influence of Be L7402 and Hep G2.These two kind of cellshad similar metabolic patterns.Pathway enrichment analysis of metabolites confirmed that arginine deprivation has grately impacted on the metabolic processes of amino acids,nucleic acids and lipids in Be L7402 and Hep G2,which suggested that arginine deprivation may cause metabolic disorders in HCC.Next,we verified that arginine deprivation activated the p38α signaling pathway and promoted the expression of downstream integrin ITGB8 as well as the migration of HCC.Therefore,we may learned that in the early stage of arginine nutrition deprivation,HCC mananged to upregulate the expression of ITGB8 by activating p38α stress response pathway,thereby escaping the direct cell injury or death caused by arginine deprivation.This study enriched the theory about the effects of arginine deprivation on HCC and added a new understanding to the drug therapy targeting arginine deprivation.