| BackgroundPrimary liver cancer is one of the most common malignant tumors in the world,and its high morbidity and high mortality greatly increase the social and economic burden.Global tumor epidemiology data for 2018 shows that there are more than 841,000 new cases of liver cancer each year,and 782,000 patients die of liver cancer [1].Therefore,actively exploring the mechanism of molecular level changes and cell signal transduction pathways during the development and progression of liver cancer provides important clues for early diagnosis and treatment of liver cancer patients,and plays an important guiding role in further improving the prognosis of liver cancer patients.The post-transcriptional function of mRNA is regulated by a variety of biological processes and is of great significance in the expression of eukaryotic genes.Among them,RNA-binding proteins(RBPs)are of great significance in the post-transcriptional regulation of mRNA [2].Abnormal expression of RBPs directly leads to changes in RNA structure and function.Some of the more than 500 RBPs that have been discovered have been shown to be involved in the development of tumors,which provides new research ideas for targeted therapy of liver cancer [3].hMex-3 is an evolutionarily conserved family of RNA-binding proteins(hMex-3A to 3D)with two homologous(K-homology,KH)RNA binding domains(RBD)[4].hMex-3A,as a member of the RNA-binding protein family hMex-3,forms a P-body with hMex-3B,which plays a biological role in degrading mRNA and inhibiting mRNA translation.A large number of studies have confirmed that hMex-3A expression in bladder cancer,gastric cancer,colon cancer and other malignant tumors increased,and has an important relationship with the occurrence and development of tumors [5-8].Therefore,we speculate that hMex-3A may play an important regulatory role in the development of liver cancer.To this end,we further explored the TCGA database and analyzed the expression of hMex-3A in liver cancer tissues.At the same time,we further explored the effects on the biological function of liver cancer cells at the in vitro level,providing new insights for the early diagnosis and treatment of liver cancer.Strategy.Objective1.Using bioinformatics technology to analyze the expression of hMex-3A in liver cancer and its relationship with the prognosis of patients with liver cancer,and to analyze its clinical significance.2.To observe the expression of hMex-3A in hepatocarcinoma tissues and adjacent tissues,and to explore the potential clinical value of hMex-3A in liver cancer.In vitro experiments were conducted to investigate the effect of hMex-3A on the biological function of hepatocarcinoma cells,and provide a theoretical basis for further exploring the molecular mechanism of hepatocarcinogenesis Methods1 The expression of hMex-3A in hepatocarcinoma was analyzed by TCGA gene expression profile data,and the relationship between hMex-3A expression and clinicopathological parameters and survival prognosis was analyzed.2.The expression of hMex-3A mRNA in hepatocarcinoma and paracancerous tissues was detected by qRT-PCR with 12 pairs of liver cancer patients and matched paracancerous tissues.Six pairs of hepatocarcinoma tissues and matched paracancerous tissues were collected and detected by Western blotting.Expression of hMex-3A protein in liver cancer tissues.3.Immunohistochemistry(IHC)was used to stain the tissue microarray containing 93 cases of liver cancer samples,and to explore the relationship between the expression level of hMex-3A and the survival prognosis of patients with liver cancer.4.By transfecting hMex-3A interfering RNA,knock down the expression of hMex-3A in hepatoma cells,and explore hMex-3A on liver cancer through cell functional experiments such as CCK-8 assay,plate cloning assay,scratch assay and Transwell assay.The effects of malignant biological functions of cells.Results1.hMex-3A is up-regulated in hepatocarcinoma and associated with clinical prognosis in patients with liver cancer The expression of hMex-3A in hepatocarcinoma tissues was higher than that in adjacent tissues(P=0.0044).The expression of hMex-3A in patients with stage III & IV liver cancer in TNM stage was higher than that in stage I & II(P=0.0308).The level of hMex-3A expression in patients with grade III & IV liver cancer was higher than that of patients with liver cancer of grade I & II(P = 0.0234).Univariate analysis indicated that high expression levels of hMex-3A(P<0.001),tumor size(P=0.021),and TNM grade(P<0.001)were associated with survival outcomes.COX multivariate analysis showed that high expression levels of hMex-3A(HR=1.491,P=0.009)and TNM stage(HR=2.298,P=0.024)were independent risk factors for clinical prognosis of patients with liver cancer.Kaplan-Meier analysis indicated that the overall survival rate of patients with high expression of hMex-3A was lower than that of patients with low expression of hMex-3A(P=0.0041).2.hMex-3A affects the malignant biological function of liver cancer cells It was confirmed that the expression levels of hMex-3A in Hep3 B and SK-Hep-1 in hepatoma cell lines were higher than those in other liver cancer cell lines.Compared with the control group,the expression of hMex-3A in hepatoma cells Hep3 B and SK-Hep-1 could significantly inhibit cell proliferation and cloning ability;the scratch test and Transwell experiment showed that the expression of hMex-3A in liver cancer cells could be down-regulated.Inhibition of migration and invasion of liver cancer cells.Conclusion1.The expression of hMex-3A mRNA and protein was up-regulated in hepatocarcinoma tissues and hepatocarcinoma cells,which was associated with the development of liver cancer.2.The high expression of hMex-3A in hepatocarcinoma is an independent risk factor for survival prognosis of liver cancer patients,and can be used as a predictor of prognosis.3.Knocking down hMex-3A can significantly inhibit the biological functions of liver cancer cells such as proliferation,cloning ability,migration and invasion,and has potential value as a therapeutic target. |
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