The Mechanism Of MiR-124-3p In The Metastasis Of Non-small Cell Lung Cancer Multi-omics To Explore The Impact Of HLTF Deletion On Hepatocellular Carcinoma

Background:Metastasis is a significant factor that affects the survival of patients with nonsmall cell lung cancer(NSCLC).Nevertheless,the molecular regulatory mechanism underlying the metastasis is currently not fully understood.This study aims to identify the important role of miR-124-3p in metastasis of NSCLC,thereby providing a potential therapeutic intervention for the NSCLC patients.Methods:Exosome secretion was determined by Nanoparticle Tracking Analysis(NTA)and the uptake was measured by fluorescence inverted microscope.The binding mechanism between miR-124-3p and its target genes was validated experimentally by Luciferase reporter.Cells migration was evaluated by Transwell assays.Transcriptome sequencing on NSCLC cells was carried out to verify the potential signaling pathway underlying miR-124-3p regulation.The protein expression levels of PI3K,p-PI3K,AKT and p-AKT in NSCLC cell lines were evaluated using Western blot analysis.The role of miR-124-3p to suppress the tumor metastasis was verified in NSCLC xenograft model.Results:Exosomes were more abundant in serum from patients with advanced lung cancer(n=24 patients)than in these from patients with early-stage lung cancer(n=30 patients),which suggested the potential correlation between amount of exosome secretion and the metastasis of NSCLC.Interestingly,the exosome release,uptake and the migration of NSCLC cells were significantly inhibited by miR-124-3p.LINC00511 suppressed the expression of miR-124-3p to facilitate exosome transport due to its role as the competitive endogenous RNA for miR124-3p.The miR-124-3p could directly target the 3’-UTR of Rab27a in NSCLC cells to inhibit exosome secretion and thereby prevent cell migration and invasion.Aside from the inhibition of exosome transport,miR-124-3p inhibited the activation of PI3K/AKT signaling in the intracellular environment.Finally,by measuring subcutaneous tumor weight and volume and lung metastasis,we also demonstrated that miR-124-3p inhibited tumor growth in vivo.Conclusion:In NSCLC,miR-124-3p significantly suppressed metastasis through extracellular exosome transport and intracellular PI3K/AKT signaling.These findings provide new insights toward a better understanding of the NSCLC metastasis and suggest a potential treatment method for NSCLC.Background:Helicase Like Transcription Factor(HLTF)is a member of the SWI/SNF family of nucleosome remodeling complexes.It has been found that HLTF is highly methylated in 40%of colorectal cancer patients and accelerated intestinal tumorigenesis suggesting that HLTF is a tumor suppressor.In addition,the HLTF gene was also highly methylated in patients with gastric and cervical cancers.However,two studies on HLTF in liver cancer showed that only 6.3%and 9.8%of patients showed promoter methylation.Therefore,the mechanism of HLTF in HCC still needs further investigation.Methods:(1)HLTF was knockout through CRISPR/Cas9 in HCC cell line,(2)RNA-seq was applied to detect and analyze the differentially expressed genes of the wild type and HLTF knockout cells,(3)ATAC-seq was applied to detect the changes in chromatin accessibility of the wild type and HLTF knockout cells,(4)The multi-omics analysis of RNA-seq and ATAC-seq was used to find the key signaling pathways and downstream genes of HLTF.Results:TCGA data base analysis showed that HLTF was highly expressed in hepatocellular carcinoma and was associated with poor prognosis,RNA-seq sequencing showed that 563 genes were up-regulated and 656 genes were down-regulated in HLTF knockout cells compared to wild-type cells,ATAC-seq analysis showed a total of 27,818 regions had significantly altered in chromatin accessibility with HLTF deletion,including 14,225 regions with enhanced chromatin accessibility and 13,593 regions with weakened chromatin accessibility,motif enrichment analysis showed that Atf3,Fral and BATF were enriched in regions with enhanced chromatin accessibility,Fra1,Fra2 and JunB were enriched in regions with weakened chromatin accessibility,the combination of RNA-seq and ATAC-seq analysis showed that the overlapping genes were mainly enriched in the Arachidonic acid metabolism pathway,Wnt signaling pathway,Calcium signaling pathway and the TGF-β signaling pathway,the deletion of HLTF increased the expression of NXF3 and highly-ressed NXF3 was associated with better prognosis in HCC patients.Conclusions:Our study pointed out that HLTF may be involved in regulating Arachidonic acid metabolism pathway,Wnt signaling pathway,TGF-β signaling pathway,etc.NXF3 may be a potential downstream gene of HLTF,and our study provided directions and ideas to elucidate the mechanism of,HLTF in hepatocellular carcinoma.