| Liver cancer is one of the most common malignant tumors with a high mortality rate.In China,the mortality rate of this disease accounts for second of all malignant tumor mortality.Among them,hepatocellular carcinoma(HCC)accounts for about 91.5%of liver cancer.At present,the main clinical treatment strategies of HCC are surgical resection,transcatheter arterial chemoembolization,and radiotherapy,gene targeting therapy,tumor ablation and liver transplantation.Although the current knowledge of diagnosis and treatment methods has been greatly advanced with the advancement of science,the long-term survival rate of patients with HCC is still low,mainly due to the high recurrence rate of liver cancer and the high rate of metastasis of cancer cells.Lead to a patient's prognosis with low survival.Therefore,further elucidation of the molecular mechanisms of HCC regulation and the exploration of its potential therapeutic targets are critical in clinical treatment.With the continuous development of human genome research programs and gene sequencing technologies,the expression of long-chain non-coding RNAs(lncRNAs)has been found in many studies on the early diagnosis and pathogenesis of human malignant tumors.In an unregulated state,lncRNA acts as a non-protein-encoding RNA transcript of more than 200 nucleotides and is a major regulator of the development and progression of human cancer,including malignant tumors such as liver cancer and gastric cancer with high global incidence.At the same time,many previous studies have shown that lncRNAs can directly or indirectly regulate the expression of tumor suppressor genes or oncogenes.Some lncRNAs have been used as biomarkers for liver cancer in patients with liver cancer to treat prognosis,diagnosis,and even predict treatment outcomes.LncRNAs show a key regulatory role in the regulation of hepatoma cell function,such as proliferation,invasion,apoptosis,metastasis and drug resistance,showing great research potential.In addition,lncRNAs circulating in the body fluids of tumor patients can be used as a non-invasive detection tool for liver cancer patients,so that the patient's personal health status can be observed at any time.In this study,the study object FEZF1-AS1(FEZ family zinc finger 1 antisense RNA 1)is a newly discovered lncRNA with a length of 2653 nucleotides located at the 7q31.32 position of the chromosome.Long non-coding RNA FEZF1-AS1 has been reported to be involved in many types of human malignancies,including colorectal cancer,gastric cancer and lung adenocarcinoma,but its function in HCC remains unclear.Therefore,in-depth study of the regulation mechanism of lncRNAs FEZF1-AS1 in liver cancer tissues has important clinical preventive and therapeutic significance.Research purposesIn the present study,we aimed to investigate the expression patterns and expression levels of FEZF1-AS1 in HCC tissues and cell lines.At the same time,the biological functions of FEZF1-AS1 during HCC progression were discovered by in vitro and in vivo assays,aiming to further elucidate the potential mechanism of lncRNA FEZF1-AS1 in regulating tumor cell proliferation,invasion,apoptosis and metastasis in hepatocellular carcinoma.Research methods(1)The second-generation sequencing technology was used to screen the significant differential genes in HCC tissues and surrounding non-tumor tissues,and the sequencing data were analyzed.The genes with significant differences were screened out,and the differential signaling pathways related to liver cancer were analyzed.RT-qPCR technique verified the difference in expression levels in IncRNA FEZF1-AS1 HCC tissues and surrounding non-tumor tissues.(2)To retrospectively analyze the correlation between the expression level of lncRNA FEZF 1-AS 1 and the overall survival and clinicopathological features of patients with liver cancer,and to elucidate the clinical significance of the difference in the expression level of lncRNA FEZF1-AS1 in HCC patients.(3)Through the use of CCK-8,flow cytometry,immunofluorescence detection of cell invasion and migration experiments and other in vitro functional detection techniques,analysis of lncRNA FEZF1-AS1 expression level changes on HCC cell proliferation,apoptosis,migration,invasion and the impact of tumor function.(4)The abnormally expressed protein was screened by gene chip analysis of the hepatoma cells knocking down/overexpressing lncRNA FEZF1-AS1,and the function of Wnt/?-catenin signaling pathway and lncRNA FEZF1-AS1 on tumor cells was further studied by immunofluorescence and immunoblotting.(5)By constructing a mouse liver cancer tumor model,the effects of up-regulated expression of ?-catenin and down-regulation of lncRNA FEZF1-AS1 on the biological function of mouse hepatocarcinoma were further investigated.Research results(1)Our study found that through the second-generation sequencing study,the expression of lncRNA FEZF1-AS1 in liver cancer tissues and surrounding non-tumor tissues was significantly different,and it was confirmed that lncRNA FEZF1-AS1 was significantly lncreased in tissues of HCC patients.(2)Retrospective analysis showed that the high expression of lncRNA FEZF1-AS1 was positively correlated with the late TNM grade,lymphoid tissue metastasis and tumor cell metastasis of liver cancer,and poor prognosis.(3)LncRNA FEZF1-AS1 up-regulated expression can promote the process of proliferation,migration and invasion of liver cancer cells,and can induce apoptosis of tumor cells,knocking down the expression results.(4)LncRNA FEZF1-AS1 expression regulation by immunofluorescence and immunoblotting experiments showed that lncRNA FEZF1-AS1 overexpression could regulate the expression level of Wnt/?-catenin signaling pathway in HCC cells,and led to its expression in the cytoplasm Experiments that accelerated the proliferation of tumor cells and knock down their expression lead to the opposite result.(5)When lncRNA FEZF1-AS1 was down-regulated,it regulated the expression of Wnt/?-catenin signaling pathway to inhibit the growth of HCC cells and promote its apoptosis.Moreover,it was also confirmed in the animal tumor formation model that the down-regulated expression of lncRNA FEZF1-AS1 can inhibit the regulation of tumor growth.ConclusionThis study demonstrated that lncRNA FEZF1-AS1 was up-regulated in HCC and affected tumor cell metastasis and prognosis.Up-regulation of lncRNA FEZF1-AS1 expression could positively regulate the expression of Wnt/?-catenin signaling pathway to regulate the biological function of tumors.It could play a regulatory role as a tumor activator,and indicate that Wnt/?-catenin signaling pathway,and lncRNA FEZF1-AS1 could be used as new prognostic biomarkers and therapeutic targets for HCC,which had clinical therapeutic and research significance. |
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