The Function And Mechanism Of Long Non-coding RNA AK058003 On Cell Proliferation And Metastasis In Hepatocellular Carcinoma Cells

Objective:Hepatocellular carcinoma(HCC)is one of the most common malignancies in the world,but its pathogenesis has not been fully elucidated so far.A large number of studies have shown that lncRNA is closely related to the occurrence and development of hepatocellular carcinoma.In order to further clarify the biological function and regulation mechanism of IncRNA in hepatocarcinoma cells,from the new angle,the followed study was performed,which provided the theoretical basis for clinical diagnosis and targeted therapy.Methods:1.The differentially expressed IncRNAs were screened from the both expression level and IncRNA sequence integrity using lncRNA expression profiles and bioinformatics software.The expression of lncRNA was confirmed by real-time quantitative PCR(q-RT-PCR)in hepatocellular carcinoma tissue,and finally we selected lncRNA-AK058003 as the research object.2.The relationship between the expression of lncRNA-AK058003 detected by qRT-PCR and the clinicopathological features and prognosis of patients with hepatocellular carcinoma resection were analysized.3.The mechanism of lncRNA-AK058003 in proliferation,invasion and migration of hepatocellular carcinoma was analyzed by in vitro and in vivo experiments.4.The target gene was predicted by bioinformatics software.The relationship between lncRNA-AK058003 and target gene was verified by RNA-Pull Down,Western blotting and qRT-PCR.5.The location of lncRNA-AK058003 was detected by FISH test,and the specific regulation between IncRNA-AK058003 and target gene was further explored.Results:1.The qRT-PCR assay of liver cancer tissues showed that IncRNA-AK058003 was lower expression in hepatocellular carcinoma.It found that IncRNA-AK058003 was associated with relapse and metastasis.2.Performed gain-of-function and loss-of-function measurements of IncRNA-AK058003,the results showed that up-regulation of IncRNA-AK058003 significantly inhibited the proliferation of hepatoma cells,and the proliferation of hepatocellular carcinoma cells after down-regulated lncRNA-AK058003 was significantly higher than that of control group(P<0.05).The results of cell cycle analysis showed that lncRNA-AK058003 significantly inhibited the migration and proliferation of hepatocarcinoma cells.The results of migration and invasion of lncRNA-AK058003 significantly inhibited the migration and proliferation of hepatocarcinoma cells.Western blot analysis showed that lncRNA-AK058003 significantly inhibited the expression of CDK4,CDK6,Cyclin D1 and MMP9,and increased the expression of E-cadherin.3.Nude mice subcutaneous tumor-bearing model and tail vein injection metastasis model found:lncRNA-AK058003 can significantly inhibit the growth and metastasis of liver cancer.4.The bioinformatics software and RNA-pull down experiments showed that IncRNA-AK058003 could bind to SNCG.LncRNA-AK058003 can also be used as a precursor of miRl 5a to inhibit the expression of SNCG mRNA and inhibit its protein expression.The expression of SNCG is reduced by the effect of HUR on the stability of SNCG and the expression of SNCG.Conclusion:It was found that lncRNA-AK058003 was closely related to the occurrence,development and metastasis of HCC,and inhibited the proliferation and metastasis of tumor by affecting HuR stabilized protein or acting as precursors of miRl 5a to inhibit the expression of SNCG.Which provides a new candidate marker for the early diagnosis and prognosis of hepatocellular carcinoma,and provides new ideas and potential targets for the personalized treatment of patients with clinical liver cancer.