Circ_0074027 contributes to non-small cell lung cancer progression through positively modulating RHOA via sequestering miR-2467-3p.

Non-small cell lung cancer (NSCLC) is a common cancer with an unfavorable 5-year survival rate. We intended to explore the role of circular RNA_0074027 (circ_0074027) in NSCLC progression. The levels of circ_0074027, messenger RNA (mRNA) of its linear form paired like homeodomain 1 (PITX1), microRNA-2467-3p (miR-2467-3p) and ras homolog family member A (RHOA) mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK8) assay and plate colony assay were conducted to measure the proliferation ability of NSCLC cells. Transwell assays were used to assess cell migration and invasion abilities. Flow cytometry was utilized to analyze cell apoptosis rate. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were used to test the interaction between miR-2467-3p and circ_0074027 or RHOA. Western blot assay was performed to evaluate the protein level of RHOA in NSCLC cells. Murine xenograft model was built to evaluate the role of circ_0074027 in tumor growth in vivo. Circ_0074027 expression was prominently elevated in NSCLC tissues and cell lines. Circ_0074027 knockdown or miR-2467-3p overexpression suppressed cell proliferation, migration and invasion and facilitated cell apoptosis of NSCLC cells. Circ_0074027 interacted with miR-2467-3p, and RHOA was a target of miR-2467-3p in NSCLC cells. RHOA silencing blocked the malignant potential of NSCLC cells. Circ_0074027 silencing restrained the malignant phenotypes of NSCLC cells largely through up-regulating miR-2467-3p. Circ_0074027 knockdown notably blocked xenograft tumor growth in vivo. In conclusion, circ_0074027 accelerated NSCLC progression by binding to miR-2467-3p to induce RHOA expression.

Identification of Key Genes Affecting Results of Hyperthermia in Osteosarcoma Based on Integrative ChIP-Seq/TargetScan Analysis.

BACKGROUND The purpose of this study was to research the effects of hyperthermia on osteosarcoma (OS) by integrating the Chromatin Immunoprecipitation with the generation sequencing (ChIP-Seq) and TargetScan analysis of heat shock transcription factor 1 (HSF1). MATERIAL AND METHODS The HSF1 ChIP-seq dataset of GSE60984 was downloaded from the Gene Expressed Omnibus (GEO) database. The HSF1-binding sites were screened by MACS2 in OS cells after 10 and 20 min of hyperthermia, and they were annotated using the ChIPseeker package. The overlapped genes were selected out when HSF1-binding sites were located in the promoter region. The Database for Annotation, Visualization, and Integrated Discovery (DAVID) was used to perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the overlaps. The miRNA-gene pairs of the overlaps were screened out via TargetScan, and the miRNA-gene-regulated network was constructed by Cytoscape software. RESULTS 1880 and 1283 genes of promoter regions were obtained in the osteosarcoma cells after 10 and 20 min of hyperthermia, respectively, and 889 of them were overlapped. The overlapped genes were enriched in 122 GO terms and 3 KEGG pathways. There were 13 657 pairs involved in the miRNA-gene regulated network of the overlaps. CONCLUSIONS Some biomarkers were identified for OS treated with hyperthermia. Moreover, some GO terms (regulation of programmed cell death and regulation of cell death) and pathways (p53 signaling pathway, methane metabolism, and viral myocarditis) might be involved.

Integrated multi-omics profiling yields a clinically relevant molecular classification for esophageal squamous cell carcinoma.

Integrated molecular analysis of human cancer has yielded molecular classification for precise management of cancer patients. Here, we analyzed the whole genomic, epigenomic, transcriptomic, and proteomic data of 155 esophageal squamous cell carcinomas (ESCCs). Multi-omics analysis led to the classification of ESCCs into four subtypes: cell cycle pathway activation, NRF2 oncogenic activation, immune suppression (IS), and immune modulation (IM). IS and IM cases were highly immune infiltrated but differed in the type and distribution of immune cells. IM cases showed better response to immune checkpoint blockade therapy than other subtypes in a clinical trial. We further developed a classifier with 28 features to identify the IM subtype, which predicted anti-PD-1 therapy response with 85.7% sensitivity and 90% specificity. These results emphasize the clinical value of unbiased molecular classification based on multi-omics data and have the potential to further improve the understanding and treatment of ESCC.

Hsa_circ_0017639 regulates cisplatin resistance and tumor growth via acting as a miR-1296-5p molecular sponge and modulating sine oculis homeobox 1 expression in non-small cell lung cancer.

Cisplatin (DDP)-induced chemoresistance is an important reason for the failure of non-small cell lung cancer (NSCLC) treatment. Circular RNAs (circRNAs) participate in the chemoresistance of diverse cancers. However, the function of hsa_circ_0017639 (circ_0017639) in the DDP resistance of NSCLC is unclear. Forty-one NSCLC samples (21 DDP-resistant samples and 20 DDP-sensitive samples) were utilized in the research. The relative expression levels of some genes were determined by real-time quantitative polymerase chain reaction (RT-qPCR). 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay for half-maximal inhibitory concentration (IC50) value of DDP and cell viability, colony formation and 5-ethynyl-2'-deoxyuridine (EDU) assays for cell proliferation, flow cytometry assay for cell apoptosis, transwell assay for cell invasion and wound-healing assay for cell migration were performed. The regulation mechanism of circ_0017639 was demonstrated by a dual-luciferase reporter assay. We observed higher levels of circ_0017639 in DDP-resistant NSCLC samples and cells. Functionally, circ_0017639 silencing decreased tumor growth and elevated DDP sensitivity in vivo and induced apoptosis, repressed proliferation, invasion, and migration of DDP-resistant NSCLC cells in vitro. Mechanically, circ_0017639 modulated sine oculis homeobox 1 (SIX1) expression via sponging microRNA (miR)-1296-5p. Also, miR-1296-5p inhibitor restored circ_0017639 knockdown-mediated impacts on cell DDP resistance in DDP-resistant NSCLCs. Furthermore, SIX1 overexpression counteracted the inhibiting impact of miR-1296-5p upregulation on DDP resistance and malignant phenotypes of DDP-resistant NSCLC cells. In conclusion, circ_0017639 conferred DDP resistance and promoted tumor growth via elevating SIX1 expression through sequestering miR-1296-5p in NSCLC, providing a new mechanism for understanding the chemoresistance and progression of NSCLC.

CDCA7 Facilitates Tumor Progression by Directly Regulating CCNA2 Expression in Esophageal Squamous Cell Carcinoma.

BACKGROUND: CDCA7 is a copy number amplified gene identified not only in esophageal squamous cell carcinoma (ESCC) but also in various cancer types. Its clinical relevance and underlying mechanisms in ESCC have remained unknown. METHODS: Tissue microarray data was used to analyze its expression in 179 ESCC samples. The effects of CDCA7 on proliferation, colony formation, and cell cycle were tested in ESCC cells. Real-time PCR and Western blot were used to detect the expression of its target genes. Correlation of CDCA7 with its target genes in ESCC and various SCC types was analyzed using GSE53625 and TCGA data. The mechanism of CDCA7 was studied by chromatin immunoprecipitation (ChIP), luciferase reporter assays, and rescue assay. RESULTS: The overexpression of CDCA7 promoted proliferation, colony formation, and cell cycle in ESCC cells. CDCA7 affected the expression of cyclins in different cell phases. GSE53625 and TCGA data showed CCNA2 expression was positively correlated with CDCA7. The knockdown of CCNA2 reversed the malignant phenotype induced by CDCA7 overexpression. Furthermore, CDCA7 was found to directly bind to CCNA2, thus promoting its expression. CONCLUSIONS: Our results reveal a novel mechanism of CDCA7 that it may act as an oncogene by directly upregulating CCNA2 to facilitate tumor progression in ESCC.

Involvement of Nucleotide-Binding and Oligomerization Domain-Like Receptors in the Intestinal Injury of Severe Acute Pancreatitis in Rats.

OBJECTIVES: The aim of the study was to observe the role of nucleotide-binding and oligomerization domain (NOD)-like receptors (NLR) in intestinal injury of severe acute pancreatitis (SAP) in rats. METHODS: Severe acute pancreatitis was induced by retrograde infusion of sodium taurocholate into the biliopancreatic duct. Rats were divided into the following 6 groups: sham operation, SAP treated with saline, and SAP treated with interleukin 1ß (IL-1ß)-converting enzyme inhibitor, killed at 6 or 12 hours after operation. Serum IL-18 and IL-1ß concentrations were measured. mRNA expression and protein levels of NOD1, NOD2, and NLRP3 in the intestine were measured. RESULTS: Severe acute pancreatitis resulted in significantly higher serum IL-18 and IL-1ß concentration, higher mRNA expression, and protein levels of NOD1, NOD2, and NLRP3 in intestine in SAP treated with saline groups compared with sham operation groups. This effect was attenuated by administration of IL-1ß-converting enzyme inhibitor. CONCLUSIONS: The NLRs, including NOD1, NOD2, and NLRP3, were involved in the intestinal injury in SAP through a caspase-1 pathway.

Exploration of the sequential gene changes in epithelial ovarian cancer induced by carboplatin via microarray analysis.

The purpose of the current study was to explore the carboplatin­induced sequential changes in gene expression and screen out key genes, which were associated with effects of carboplatin on epithelial ovarian cancer (EOC). The microarray dataset GSE13525 was downloaded from the Gene Expression Omnibus database, including 6 EOC cell samples separately treated with carboplatin at 24, 30 and 36 h (case group), and 6 samples treated with phosphate­buffered saline at the same time points (control group). A total of 3 sets of differentially expressed genes (DEGs) were respectively identified in case samples at 24, 30 and 36 h compared with the control group via the Limma package, and separately recorded as DEG­24, DEG­30 and DEG­36. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the overlapped DEGs were performed via the Database for Annotation, Visualization and Integrated Discovery. The protein­protein interaction (PPI) network was constructed and analyzed by Cytoscape software. In addition, the survival curves were drawn to illustrate the association between the expression levels of certain critical genes and the prognosis of EOC. A total of 170, 605 and 1043 DEGs were separately obtained in DEG­24 DEG­30 and DEG­36, and 110 overlaps were identified. The overlaps were enriched in 77 GO terms and 3 KEGG pathways. A total of 152 pairs were involved in the PPI network, and the abnormal expression levels (high or low) of c­Jun and cyclin B1 (CCNB1) would reduce the survival time of patients with EOC. The study indicated that c­Jun and CCNB1 may be the prognostic biomarkers of EOC treated with carboplatin, and certain pathways (such as p53 signaling pathway, cell cycle and mitogen­activation protein kinase signaling pathway) may be involved in carbo-platin­resistant EOC.