ABSTRACT
INTRODUCTION: The diagnosis and treatment of osteoporosis, a frequent age-related metabolic bone disorder, remain incomprehensive and challenging. The potential regulatory role of lncRNA XIST and sphingosine kinase 1 (SPHK1) pathway need experimental investigations. MATERIALS AND METHODS: RAW264.7 cells and BMMs were obtained for in vitro studies and 30 ng/mL RANKL was implemented for induction of osteoclast differentiation. The suppressing of lncRNA XIST, SPHK1 and fused in sarcoma (FUS) was achieved using small hairpin RNA, while overexpression of XIST and FUS was constructed by pcDNA3.1 vector system. Tartrate-resistant acid phosphatase (TRAP) staining was used for observation of formation of osteoclasts. RNA-pulldown analysis and RNA binding protein immunoprecipitation (RIP) was implemented for measuring mRNA and protein interactions. RT-qPCR was conducted to determining mRNA expression, whereas ELISA and Western blotting assay was performed for monitoring protein expression. RESULTS: RANKL induced osteoclast differentiation and upregulated expression of osteoclastogenesis-related genes that included NFATc1, CTSK, TRAP and SPHK1 and the level of lncRNA XIST in both RAW264.7 cells and BMMs. However, knockdown of lncRNA XIST or suppressing SPHK1 significantly reserved the effects of RANKL. LncRNA XIST was further demonstrated to be interacted with FUS and increased the stability of SPHK1, indicating its ability in promoting osteoclast differentiation through SPHK1/S1P/ERK signaling pathway. CONCLUSION: LncRNA XIST promoted osteoclast differentiation via interacting with FUS and upregulating SPHK1/S1P/ERK pathway.
Subject(s)
Bone Resorption , Osteoclasts , Proprotein Convertases/metabolism , RNA, Long Noncoding , RNA-Binding Protein FUS/metabolism , Serine Endopeptidases/metabolism , Animals , Bone Resorption/metabolism , Cathepsin K/metabolism , Cell Differentiation , Hematopoiesis , Mice , NFATC Transcription Factors/metabolism , Osteoclasts/cytology , Osteogenesis , Phosphotransferases (Alcohol Group Acceptor)/metabolism , RANK Ligand/metabolism , RAW 264.7 Cells , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Tartrate-Resistant Acid Phosphatase/metabolismABSTRACT
OBJECTIVE: To analyze the change of EB virus VCA/IgA and EA/IgA titer during the development of nasopharyngeal carcinoma (NPC), and the role in screening for NPC. METHODS: VCA/IgA and EA/IgA were monitored in a period of 12 years by immunoenzymatic titration from the sera of 54 NPC patients after primary serological screening. RESULTS: VCA/IgA and EA/IgA titer had shown gradual increment 1 - 7 years before NPC was pathologically diagnosed. The mean titer of VCA/IgA was 1:21.04, 7 - 4 years before diagnosis. VCA/IgA titer ascended quickly within 3 years before diagnosis. The geometric mean titer (GMT) of VCA/IgA and EA/IgA were 1:76.86 and 1:6.49 when NPC was diagnosed, which descended quickly after radiotherapy and, in 4 years, approached the average titer of VCA/IgA positive population. CONCLUSION: VCA/IgA titer rises uninterruptedly 3 years before NPC is diagnosed pathologically in most patients but their EA/IgA titer rises slowly. The detection of VCA/IgA titer can be used to find early NPC, whereas EA/IgA can not. The pre-clinical phase of NPC is 3 years according to this dynamic study.
Subject(s)
Antibodies, Viral/blood , Antigens, Viral/immunology , Capsid Proteins/immunology , Immunoglobulin A/blood , Nasopharyngeal Neoplasms/virology , Adult , Early Detection of Cancer , Humans , Middle Aged , Nasopharyngeal Neoplasms/diagnosisABSTRACT
OBJECTIVE: To observe the character of Epstein-Barr(EB) virus serology, fibroscopy appearance and prognosis of asymptomatic nasopharyngeal carcinoma(NPC). METHODS: Viral capsid antigen's IgA (VCA/IgA) of EB virus and early antigen's IgA(EA/IgA) of EB virus were detected by immunoenzymatic method. The clinical examination was carried out, including indirect mirror examination and fibroscopy of the nasopharynx and multiple biopsies. All patients of NPC were followed up to the end of 1999. RESULTS: 1. The geometric mean titer of VCA/IgA and EA/IgA are 1:100.79 and 1:10.76 respectively when asymptomatic NPC was diagnosed. There were no significant difference between VCA/IgA and EA/IgA antibody titres of asymptomatic patients and symptomatic cases (P > 0.05). The survival rates in these asymptomatic cases were higher than symptomatic patients (P < 0.05). 2. There was no correlation with the VCA/IgA or EA/IgA titer and the prognosis (P > 0.05) and the cervical lymph node metastasis (P > 0.05) when NPC was diagnosed. CONCLUSION: This is helpful to detect asymptomatic NPCs by EB serological screening periodically and nasopharyngeal fibroscopy and multiple biopsies.
