RESUMO
BACKGROUND: Few studies showed the changes in cytokine profiles after infection by hepatitis B virus (HBV), the most common viral liver disease worldwide. This study examined the relationship between interleukin (IL)-32, IL-1, and interferon (IFN)-γ levels and HBV load. METHODS: IL-32, IL-1, and IFN-γ levels in hepatitis B patients serum and HBV-stimulated PBMCs were measured by ELISA. Gene transcripts in PBMCs from hepatitis B patients and HBV-stimulated PBMCs from healthy controls were measured by real-time PCR. RESULTS: IL-32, IL-1, and IFN-γ protein levels in serum from hepatitis B patients were significantly higher than those in healthy volunteers (P<0.05). Hepatitis B patients showed significantly higher expression of IL-32, IL-1, and IFN-γ transcripts than healthy volunteers (P<0.05). IL-32, IL-1, and IFN-γ levels in PBMCs stimulated by different amounts of HBV were significantly higher than those in HBV-unstimulated PBMCs (P<0.05). Real-time PCR results were consistent with the ELISA results. CONCLUSIONS: The levels of IL-32, IL-1, and IFN-γ protein and transcripts in serum and PBMCs from hepatitis B patients were higher than those in healthy volunteers. Similarly, both were higher in PBMCs from healthy volunteers stimulated by HBV in vitro. However, the changes in cytokine levels were not proportional to the viral load.
Assuntos
Hepatite B Crônica/imunologia , Interferon gama/sangue , Interleucina-1/sangue , Interleucinas/sangue , Leucócitos Mononucleares/imunologia , Carga Viral , Adolescente , Adulto , Idoso , DNA Viral/genética , Ensaio de Imunoadsorção Enzimática , Feminino , Voluntários Saudáveis , Vírus da Hepatite B , Humanos , Interferon gama/genética , Interleucina-1/genética , Interleucinas/genética , Leucócitos Mononucleares/virologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Adulto JovemRESUMO
PURPOSE: The aim of our study was to validate the sway of long noncoding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) on the metabolism and growth of bladder cancer cells by microRNA-31 (miR-31)/cyclin-dependent kinase 1 ( CDK1). METHODS: The Gene Expression Omnibus database was used for analyzing the differentially expressed lncRNA and messenger RNA (mRNA) in bladder cancer tissues, with the highly expressed lncRNA PVT1 and mRNA CDK1 screened out. The expression level of PVT1 was detected by quantitative reverse-transcription polymerase chain reaction, cell viability by Cell Counting Kit-8 assay, cell proliferation and scratch by 5-bromo-2'-deoxyuridine assay, cell migration and invasion by transwell assays, the expression level of CDK1 by immunohistochemistry and western blot analysis, transcription factor targeting by dual-luciferase assay, and the effect of PVT1 on bladder cancer growth by nude mice tumor formation experiment. RESULTS: LncRNA PVT1 and mRNA CDK1 had a higher expression in bladder cancer cells than that in neighboring tissues. Activity, proliferation, colony formation, migration, and invasion of bladder cancer cell were noticeably reduced by the PVT1 inhibitor than that of control group. PVT1 and CDK1 have binding sites with miR-31. When miR-31 decreased, CDK1 mRNA and protein levels increased in vivo experiments in nude mice. When PVT1 was downregulated, the tumor size was significantly reduced and tumor proliferation was curbed. Immunohistochemistry showed that the positive rate of CDK1 and Ki-67 decreased and the expression of miR-31 increased after PVT1 was inhibited. CONCLUSIONS: LncRNA PVT1 was overexpressed in bladder cancer cells, and it was downregulated miR-31 to enhance CDK1 expression and facilitate bladder cancer cells proliferation, migration, and invasion.