UBE2C as an Immune-Related Biomarker for Breast Cancer:A Study Based on Multiple Databases

Objective To screen the target gene UBE2C and explore its prognostic value and immune correlation in breast cancer(BRCA)using multiple databases..Methods The microarray expression datasets of BRCA were downloaded from the Gene Expresssion Omnibus database(GEO)and analyzed to obtain differentially expressed genes(DEGs).Hub genes were obtained by constructing and visualizing the protein-protein interaction network of DEGs.Then the key gene UBE2C was determined using R language,STRING,and Cytoscape,and the differential expression of UBE2C was verified using the external datasets,The Cancer Genome Atlas(TCGA),and quantitative real-time PCR(qRT-PCR).The prognostic value and immunological correlation of UBE2C in BRCA were explored using R language,TIMER,and Gene Set Enrichment Analysis(GSEA).Results The expression of UBE2C was differentially upregulated in BRCA,as verified by TCGA and qRT-PCR.Prognostic analysis revealed that UBE2C served as an independent prognostic factor.High expression of UBE2C was associated with decreased immune infiltration levels of B cells,CD4+T cells,CD8+T cells,macrophages,and myeloid dendritic cells in BRCA tissue.The expression of UBE2C in BRCA showed a significant correlation with PDCD1,CD274,and CTLA4 expressions.There was a positive correlation between the expression of UBE2C and the tumor mutational burden and microsatellite instability.GSEA demonstrated that UBE2C expression significantly enriched 786 immune-related gene sets.Conclusions UBE2C expression in BRCA tissues can predict the survivals and prognosis of BRCA patients.Also,it is closely related to the BRCA immune microenvironment and can predict the effecacy of immunotherapy in BRCA patients.Therefore,UBE2C may be an potential immune-related prognostic biomarker for BRCA.

Wnt5a participates in hepatic stellate cell activation observed by gene expression profile and functional assays

AIM:To identify differentially expressed genes in quiescent and activated hepatic stellate cells(HSCs)and explore their functions.METHODS:HSCs were isolated from the normal Sprague Dawley rats by in suit perfusion of collagenase and pronase and density Nycodenz gradient centrifugation.Total RNA and mRNA of quiescent HSCs,and cultureactivated HSCs were extracted,quantified and reversely transcripted into cDNA.The global gene expression profile was analyzed by microarray with Affymetrix rat genechip.Differentially expressed genes were annotated with Gene Ontology(GO)and analyzed with Kyoto encyclopedia of genes and genomes(KEGG)pathway using the Database for Annotation,Visualization and Integrated Discovery.Microarray data were validated by quantitative real-time polymerase chain reaction(qRTPCR).The function of Wnt5a on human HSCs line LX-2 was assessed with lentivirus-mediated Wnt5a RNAi.The expression of Wnt5a in fibrotic liver of a carbon tetrachloride(CCl4)-induced fibrosis rat model was also analyzed with Western blotting.RESULTS:Of the 28 700 genes represented on this chip,2566 genes displayed at least a 2-fold increase or decrease in expression at a P<0.01 level with a false discovery rate.Of these,1396 genes were upregulated,while 1170 genes were downregulated in culture-activated HSCs.These differentially expressed transcripts were grouped into 545 GO based on biological process GO terms.The most enriched GO terms included response to wounding,wound healing,regulation of cell growth,vasculature development and actin cytoskeleton organization.KEGG pathway analysis revealed that Wnt5a signaling pathway participated in the activation of HSCs.Wnt5a was significantly increased in cultureactivated HSCs as compared with quiescent HSCs.qRTPCR validated the microarray data.Lentivirus-mediated suppression of Wnt5a expression in activated LX-2 resulted in significantly impaired proliferation,downregulated expressions of typeⅠcollagen and transforming growth factor-β1.Wnt5a was upregulated in the fibrotic liver of a CCl4-induced fibrosis rat model.CONCLUSION:Wnt5a is involved in the activation of HSCs,and it may serve as a novel therapeutic target in the treatment of liver fibrosis.

Molecular Cloning, Characterization and Sequence Analysis of KCNQ4 in Large Odorous Frog, Odorrana graminea

Acoustic communication is essential for anuran survival and reproduction, and masking background noise can affect the effective acoustic communication. The larger odorous frog(Odorrana graminea) inhabits noise montane streams, and it has shown an ultrasound communication adaptation. However, the molecular mechanism underlying their ultrasonic hearing adaptation remains unknown. To characterize and investigate the molecular characteristics and evolution of the high-frequency hearing-sensitive gene(KCNQ4) in O. graminea, termed as OgKCNQ4, the rapid amplification of cDNA ends(RACE) was performed to amplify the cDNA of OgKCNQ4. Different bioinformatics analyses were used to investigate the molecular characteristics. Multiple nucleotide and amino acid sequence alignment were conducted, and phylogenies were reconstructed under the maximum likelihood and Bayesian approaches. The full-length cDNA of OgKCNQ4 was 2065 bp, and the open reading frame(ORF) was 2046 bp encoding for a putative protein with 681 amino acids. The relative molecular weight of OgKCNQ4 was 76.453 kD and the putative PI was 9.69. Secondary structure prediction analyses suggested 42.29% alpha helixes and 43.76% random coils in OgKCNQ4. Gene homology and Phylogenetic analyses revealed the closest relationship between OgKCNQ4 and KCNQ4 of Nanorana parkeri with 96.9% similarity and 95.0% identity. We first determined the full-length cDNA of OgKCNQ4 and the results here could provide foundations for further study on the evolution of KCNQ4 and its relationship to ultrasonic communication in amphibians.