hes1-protein--human and Intervertebral-Disc-Degeneration

The objective of the study was to investigate how inflammatory cytokines, IL-1β, and TNF-α control NOTCH signaling activity in nucleus pulposus (NP) cells. An increase in expression of selective NOTCH receptors (NOTCH1 and -2), ligand (JAGGED2), and target genes (HES1, HEY1, and HEY2) was observed in NP cells following cytokine treatment. A concomitant increase in NOTCH signaling as evidenced by induction in activity of target gene HES1 and HEY1 promoters and reporter 12xCSL was seen. Moreover, treatment increased activity of a 2-kb NOTCH2 promoter. Treatment of cells with NF-κB and MAPK inhibitors abolished the inductive effect of cytokines on NOTCH2 promoter and its expression. Gain and loss-of-function studies confirmed the inductive effect of p65 on NOTCH2 promoter activity. In contrast, p50 blocked the cytokine induction of promoter activity. Supporting promoter studies, lentiviral delivery of sh-p65, and sh-IKKβ significantly decreased cytokine dependent change in NOTCH2 expression. Interestingly, MAPK signaling showed an isoform-specific control of NOTCH2 promoter; p38α/β2/δ, ERK1, and ERK2 contributed to cytokine dependent induction, whereas p38γ played no role. Analysis of human NP tissues showed that NOTCH1 and -2 and HEY2 expression correlated with each other. Moreover, expression of NOTCH2 and IL-1β as well as the number of cells immunopositive for NOTCH2 significantly increased in histologically degenerate discs compared with non-degenerate discs. Taken together, these results explain the observed dysregulated expression of NOTCH genes in degenerative disc disease. Thus, controlling IL-1β and TNF-α activities during disc disease may restore NOTCH signaling and nucleus pulposus cell function.

Topics: Adult; Aged; Animals; Basic Helix-Loop-Helix Transcription Factors; Cell Cycle Proteins; Cell Line; Female; Homeodomain Proteins; Humans; Interleukin-1beta; Intervertebral Disc; Intervertebral Disc Degeneration; Male; MAP Kinase Signaling System; Middle Aged; Promoter Regions, Genetic; Rats; Receptor, Notch1; Receptor, Notch2; Repressor Proteins; Transcription Factor HES-1; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Long noncoding RNA (lncRNA) represents a new group of transcripts which act a critical role in various biological and pathological processes. Growing evidence suggested that a new lncRNA, FAM83H-AS1, played important roles in several cancers. However, the underlying mechanisms of FAM83H-AS1-regulating functions in intervertebral disc degeneration (IDD) have yet to be explained. Thus study examined the role of lncRNA FAM83H-AS1 in progression of IDD. First, we proved that expression level of FAM83H-AS1 was expressed in nondegenerated nucleus pulposus (NP) tissues and degenerative NP samples. Moreover, we studied the expression level of FAM83H-AS1 relationship of the clinical disc degeneration grade. Our data suggested that FAM83H-AS1 expression was downregulated in normal NP samples compared with in the degenerated NP samples. FAM83H-AS1 expression was positively correlated with degree of disc degeneration grade. The expression of FAM83H-AS1 was positively correlated with scores of Pfirrmann grade. FAM83H-AS1 expression was increased by IL-1β and TNF-αtreatment in NP cells. Ectopic expression of FAM83H-AS1 induced cell growth and modulated extracellular matrix (ECM) expression in the NP cell. Elevated expression of FAM83H-AS1 promoted Notch1 and Hes1 expression in NP cells. Furthermore, FAM83H-AS1 induced NP cell growth and modulated ECM expression through targeting Notch 1. To conclude, dysregulated expression of FAM83H-AS1 played a crucial role in progression of IDD.

Topics: Cell Proliferation; Extracellular Matrix; Gene Expression Regulation; Humans; Intervertebral Disc Degeneration; Nucleus Pulposus; Receptors, Notch; RNA, Long Noncoding; Signal Transduction; Transcription Factor HES-1; Tumor Necrosis Factor-alpha