| Background and purpose:Liver cancer is one of the most common malignant tumors with poor survival and high lethality.Radiofrequency ablation(RFA),as a first-line treatment for early liver cancer,has greatly improved the prognosis and living standards of patients.However,related studies have found that tumor residues after RFA caused by various reasons are one of the important reasons for its high recurrence rate.Long non-coding RNA(lnc RNA)can participate in regulating related biological characteristics by regulating gene expression at various levels.However,the role and mechanism of lnc RNAs in the rapid progression of residual lesions after RFA in liver cancer remains unclear.Materials and Methods:1.In vivo simulation of the formation of residual foci after incomplete radiofrequency ablation of liver cancer: Hep G2,a liver cancer cell,was implanted subcutaneously in nude mice,and radiofrequency ablation was performed after culture.Hematoxylin-eosin staining(HE)staining was used to determine the pathological morphology after ablation,and mitochondrial enzyme activity assay was used to evaluate tumor tissue activity after ablation.2.Screen differentially expressed lnc RNAs and mRNAs in residual tumor tissues: Through high-throughput sequencing of residual tumor tissues and control tissues,select differentially expressed lnc RNAs and mRNAs in residual tumor tissues.3.Functional analysis of differential lnc RNAs and mRNAs: A lnc RNA-mRNA co-expression network was constructed by correlation analysis.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were used to determine the function of differentially expressed mRNAs in residual tumor tissue.4.Expression and function analysis of lnc RNA ASB16-AS1 in liver cancer:Bioinformatics analysis of expression of ASB16-AS1 in liver cancer.After si RNA-ASB16-AS1-si-1 was transfected into MHCC97 H and MHCC97 L cells,respectively,the effect of cell proliferation was examined by CCK-8 experiment.5.Analysis of the mechanism of action of ASB16-AS1 in liver cancer:explore the possible mechanism of ASB16-AS1 affecting the biological activity of residual tumors through expression correlation,transcription factor binding sites and structural fitting.Result:1.Residue formation after incomplete radiofrequency ablation treatment of liver cancer in vivoThe average diameter of subcutaneous tumors of liver cancer in nude mice after culture is about 1.5x1.5cm.After RFA treatment,the results of HE staining showed that the ablation center was a single uniform red stain,and the fragmented cell nuclear structure was visible in the surrounding area.The measurement of mitochondrial enzyme activity showed the existence of tumor tissue activity around the ablation.2.Screen differentially expressed lnc RNAs and mRNAs in residual tumor tissuesThrough high-throughput sequencing technology detection,a total of 161 lnc RNAs were highly expressed and 579 were low-expressed in the residual tumor tissue after radiofrequency ablation.In addition,87 highly expressed mRNAs and 576 lowly expressed mRNAs were found in residual tumor tissues.3.Functional analysis of differentially expressed lnc RNAs and mRNAsA total of 1201 co-expression correlations between lnc RNAs and mRNAs were collected.The GO and KEGG pathway analysis of up-and down-regulated genes,respectively,showed that differential mRNA was significantly enriched in antigen processing,endogenous antigen presentation,regulation of cell metabolic processes,MAPK signaling and cell cycle regulation-related pathways.in.4.ASB16-AS1 is highly expressed in liver cancer and regulates the proliferation of liver cancer cellsASB16-AS1 was highly expressed in residual cancer tissues by chip and GEO expression data analysis.Based on the analysis of TCGA large samples,lnc RNA ASB16-AS1 was more highly expressed in liver cancer than normal liver tissue(p <0.01).The results of CCK-8 experiments showed that compared with the control group,the activity of hepatoma cells MHCC97 H and MHCC97 L decreased after ASB16-AS1 silencing.5.Analysis of the role of ASB16-AS1 in liver cancerCorrelation analysis of residual cancer sequencing data,TCGA data and GEO data showed that the transcription factor related to the expression of ASB16-AS1 was FOXP4.At the same time,the mRNAs associated with the expression of FOXP4 are NDST2,ERAL1,CCDC85 C,and SLC39A13.The results of molecular docking showed that the binding score of FOXP4 and ASB16-AS1 was-355.65,and the binding score of FOXP4 and NDST2 was-323.43.Conclusion:1.Successfully constructed a residual tumor tissue model after incomplete radiofrequency ablation of liver cancer,in which the expression of lnc RNAs and mRNAs in the residual cancer tissue changed.2.Bioinformatics analysis confirmed that differentially expressed mRNAs play an important role in antigen presentation,cell metabolism,signal transduction and cell cycle.3.ASB16-AS1 is highly expressed in residual tumor tissues and promotes the proliferation of liver cancer cells.4.ASB16-AS1 may regulate the expression of NDST2 by binding to the transcription factor FOXP4,thereby participating in regulating the proliferation capacity of residual cancer after radiofrequency ablation treatment of liver cancer. |
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