LCMT1 Promotes The Proliferation Of Hepatocellular Carcinoma Cells Through Glycolysis

Background: Hepatocellular carcinoma(HCC)is an extraordinary heterogeneous disease of the liver in primary malignancy cancers.The curative effect of HCC is poor and the mortality is high due to its high degree of malignancy and rapid development.In recent years,the clinical application of liver cancer chemotherapy drugs such as Sorafenib and Donafenib has improved the prognosis of liver cancer patients to a certain extent,but due to the emergence of toxicity and drug resistance,the application of chemotherapy drugs also has its limitations.Therefore,it is of great significance to understand the specificity of the changes in molecular biology during the occurrence and development of liver cancer and to carry out effective prevention,diagnosis and treatment of liver cancer patients.Aerobic glycolysis,a significant metabolic process in tumor cells,is characterized by increased glucose utilization and lactic acid production.Targeting tumor metabolism has always been an important development direction in tumor therapy.LCMT1 is the only methyltransferase of protein phosphatase 2A(PP2A),which plays an important role in regulating the assembly and activity of PP2 A holoenzyme.Previous studies in vivo and in vitro have shown that LCMT1 can catalyze PP2 A methylation to regulate the gluconeogenic pathway to maintain glucose homeostasis in the liver.Objective: A small cohort of HCC tissue samples and an online HCC tumor database were used to investigate the role of LCMT1 in tumor genesis and progression,and to explore whether LCMT1 may regulate tumor growth by glycolysis.Methods&Resluts: 1.The postoperative tumor tissues and paracancerous tissues of patients with hepatocellular carcinoma from a hospital from December2018 to June 2019 were collected,and the expression level of LCMT1 was detected by fluorescence quantitative PCR and western blot.2.The results of forest tree analysis by Mate showed that the expression level of LCMT1 in liver tissues of HCC patients in GEO database was higher than that in adjacent tissues,and there was no publication bias.3.Clinical baseline data of patients with HCC were obtained on the online analysis website of c Bio Portal integrated tumor database.The results showed that the different expression levels of LCMT1 were different among Asian ethnic group(p=0.008),histological grade(p=0.006),tumor size(p=0.016)and family history(p=0.010),and were positively correlated with histological grade and tumor size.The higher the expression level of LCMT1,the higher the histological grade and the larger the tumor.However,there were no significant differences in other clinical features such as age,gender,alcohol consumption,HBV infection,T stage and vascular invasion(p>0.050).K-M survival curve analysis showed that the overall survival time and disease-free survival time of patients with high expression of LCMT1 were relatively short,and the difference was statistically significant(p< 0.010).4.The results of CCK8 and Ed U proliferative detection kit showed that silencing LCMT1 inhibited the proliferation of Hep G2 and Huh7 hepatoma cells,while over expression of LCMT1 promoted Hep3 B cells.q-RTPCR results showed that silencing of LCMT1 decreased the expression of proliferation-related protein Mk I67,while over expression of LCMT1 promoted the expression of proliferation-related protein Mk I67.colony formation experiments showed that silencing LCMT1 reduced the ability of colony formation of Hep G2 and Huh7 hepatoma cells.The scratch experiment showed that silencing LCMT1 did not affect the migration ability of Huh7 cells.5.Silencing LCMT1 decreased glucose utilization,lactic acid production,activities of LDH and PK in Hep G2 and Huh7 cells,and m RNA expression levels of GLUT1,HK2,PFK2 and MCT4.Over expression of LCMT1 increased glucose utilization and lactic acid production in Hep3 B cells,increased the activities of lactate dehydrogenase and pyruvate dehydrogenase,and promoted the expression levels of GLUT1,HK2,PFK2 and MCT4 m RNA.6.The expression level of LCMT1 m RNA in tumor tissues of hepatocellular carcinoma patients in TCGA database is related to the regulatory genes of glycolysis.There were significant positive correlations among GLUT1,HK2,PFK2,PK,LDH and MCT4 m RNA expression levels(p< 0.05).Concluction: 1.LCMT1 is highly regulated in hepatocellular carcinoma with poor prognosis.2.LCMT1 promotes the growth and proliferation of HCC cells lines.3.LCMT1 is involved in the growth of liver cancer by promoting glycolysis.