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Objective:To determine the role and molecular mechanism of dexmedetomidine (DEX) in postmenopausal osteoporosis (PMOP) .Methods:Twenty-seven patients with PMOP admitted to Yantai Yantaishan Hospital from Jan. 2020 to Jan. 2021 were selected as PMOP group, and 20 healthy volunteers were selected as Normal group. The differentially expressed miRNAs in PMOP were screened, clinically, the expression of miR-483-3p and catenin beta 1 (CTNNB1) in serum samples of patients with PMOP was detected by qRT-PCR. In vitro experiment, Bone marrow mesenchymal stem cells (BMSCs) were induced into osteoblasts, Dex was used to treat BMSCs and intervene the expression of miR-483-3p, CTNNB1 in BMSCs, the expression level of osteogenesis related indexes (RUNX2、OCN、OPN) was detected. After coculturing Human umbilical vein endothelial cell (HUVECs) with BMSCs, angiogenesis experiment was utilized to detect the angiogenesis ability.Results:Compared with Normal group (1±0.46) (1.03±0.44) , the expression of miR-483-3p (3.23±1.61) was increased in serum of PMOP patients while expression of CTNNB1 (0.50±0.27) was inhibited ( t=5.99, P<0.001) ( t=5.14, P<0.001) . miR-483-3p has a good diagnostic effect on PMOP (AUC=0.86, P<0.001) . After Dex treatment, miR-483-3p level was decreased in BMSCs, CTNNB1 level was increased (all P<0.05) . Dex promoted the expression of RUNX2, OCN, OPN and number of angiogenesis, but this effect was partially reversed by miR-483-3p overexpression (all P<0.05) . CTNNB1 was confirmed as a target gene of miR-483-3p, the inhibition effects of miR-483-3p overexpression on osteogenic differentiation and angiogenesis of BMSCs induced by Dex was partially reversed by CTNNB1 overexpression (all P<0.05) . Conclusion:Dex enhanced CTNNB1 level in PMOP via inhibiting miR-483-3p, subsequently promoted osteogenic differentiation and angiogenesis of BMSCs and inhibited progression of PMOP.
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AIM:To investigate the effect of miR-483-5p on P3 promoter-driven mRNA(P3 mRNA)expres-sion of human insulin-like growth factor 2(IGF2)gene and its role in the development of hepatocellular carcinoma (HCC).METHODS: The expression levels of miR-483-5p and P3 mRNA were analyzed by real-time PCR in human HCC cell lines Huh7,Hep3B,Bel-7402,HepG2 and SMMC-7721,normal human liver cell line HL-7702,83 cases of hu-man HCC tissues and their matched adjacent nontumorous tissues(MANT), and 22 cases of normal adult liver tissues (NALT).The association between P3 mRNA level and miR-483-5p level was evaluated by Pearson correlation analysis. The full-length sequences of 5'UTR of P3 mRNA containing wild-type and mutant miR-483-5p-binding sequences were cloned into pGL3 promoter vector, which were termed pGL3-P3-5'UTR-WT and pGL3-P3-5'UTR-MUT, respectively. These luciferase reporter constructs were transfected into HeLa,293T and Huh7 cells together with miR-483-5p mimic, miR-483-5p inhibitor or scrambled control,and the luciferase activity was measured using dual-luciferase reporter system. The miR-483-5p mimic,miR-483 inhibitor and scrambled control were also transfected into Huh 7 cells and Hep3B cells, and P3 mRNA level was detected by real-time PCR.The expression levels of miR-483-5p in the nuclear and cytoplasmic fractions of Hep3B cells and Huh7 cells were detected by real-time PCR.The effect of miR-483-5p on P3 mRNA transcrip-tion was evaluated by nuclear run-on assay.The effect of miR-483-5p on the stability of P3 mRNA was analyzed by RNA stability assay.Furthermore,the effects of miR-483-5p on the viability, apoptosis, migration and invasion of Huh7 cells were investigated.RESULTS:Significamtly high levels of miR-483-5p and P3 mRNA were detected in the 5 human HCC cell lines and the human HCC tissues as compared with the human normal liver cell line HL-7702, and the MANT and NALT,respectively.Linear correlation analysis revealed that P 3 mRNA level was positively correlated to miR-483-5p level in the 5 human HCC cell lines and the human HCC tissues.miR-483-5p directly recognized the P3 mRNA 5'UTR to pro-mote gene expression.Overexpression of miR-483-5p resulted in a significant increase in P3 mRNA expression in a dose-dependent manner in the Huh7 cells and Hep3B cells.The mature miR-483-5p was present in both cytoplasm and nucleus of Hep3B cells and Huh7 cells.miR-483-5p induced nascent P3 mRNA transcription in the nucleus of Huh7 cells.miR-483-5p did not alter P3 mRNA stability in Huh7 cells.Furthermore,miR-483-5p led to increased viability,apoptosis inhi-bition,and enhanced migration and invasion abilities in the Huh 7 cells.CONCLUSION:High expression of miR-483-5p promotes the growth,migration and invasion of HCC cells in part through up-regulating P3 mRNA transcription,and is con-sequently involved in the development of HCC.
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Objective To investigate the effects of mir?483 on the proliferation,invasion and migration of nasopharyngeal carcinoma cells . Methods RT?qPCR was used to detect mir?483 expression level in different nasopharyngeal carcinoma cells(CNE?1,CNE?2)and the alteration of mir?483 expression level in these cells after radiation. CNE?1,one of the NPC cells,was selected. Cationic liposomes transient transfection was used to construct cells which overexpressed mir?483 or negative control . MTT assay was conducted to test the cell proliferation ability. Wound scratch assay and transwell migration assay were used to detect the effects of mir?483 on NPC invasion and migration ability. Results In terms of radiation resistance ,the expression level of mir?483 was higher in CNE?1. Overexpressing mir?483 can enhance the proliferation ,invasion and migration ability of CNE?1. Conclusions Mir?483 enhances the proliferation,invasion and migration ability of nasopharyngeal carcinoma cells.
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Objective To investigate the expression level of serum miR-193a-3p,miR-337-5p and miR-483-5p in esophageal squamous cell carcinoma (ESCC) patients and to explore their value for diagnosis and prognosis of ESCC.Methods Serum samples were collected from 63 ESCC patients before and after surgery in Nanjing General Hospital and Xuzhou Cancer Hospital between June 2013 and May 2014 and serum sanples of 63 age-and sex-matched healthy individuals acted as the normal controls.TaqMan Low Density Assay was used to detect the deregulated miRNA in ESCC patients and then quantitative real-time PCR was used to validate the upregulated miRNA miR-193a-3p,miR-337-5p and previously reported miR483-5p that was upregulated in oral squamous cell carcinoma (OSCC).Finally,the three miRNA were evaluated for their clinical value in the diagnosis and predicting prognosis of ESCC.Results Compared with normal controls,serum levels of miR193a-3p,miR-337-5p and miR-483-5p in ESCC patients were significantly up-regulated (0.459±0.339 vs 0.195±0.084,U =591;5.686±5.211 vs 2.476±0.808,U=605;32.545 ± 22.479 vs 19.509±10.601,U=1 037,respectively,all P＜ 0.0001) and their levels were significantly reduced after the surgical treatment (P＜0.05).The areas under the ROC curve of serum miR-193a-3p,miR-337-5p,miR-483-5p and miR-Panel were all larger than that of CEA.Univariate Kaplan-Meier analysis revealed that ESCC patients with low expression level of miR-483-5p in postoperative serum exhibited higher survival rate than those with high level(P=0.022).Conclusion Serum miR-193a-3p,miR-337-5p and miR-483-5p can be potential molecular biomarkers in the diagnosis and predicting prognosis of ESCC.
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Background and purpose:Although cisplatin-based chemotherapies are used as the first-line treatment for ovarian cancers, the majority of patients eventually progress with platinum-resistant disease. miR-483-5p is overexpressed in lung cancer. However, the research on miR-483-5p in epithelial ovarian cancer (EOC) is still unclear. This study aimed to investigate the expression of miR-483-5p in EOC and its effects on cisplatin resistance in EOC cells.Methods:This study analyzed the expression of the miR-483-5p by real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) in EOC tissues, normal ovarian tissues, and EOC cells. The role of miR-483-5p in EOC was evaluatedin vitro by lentivirus-mediated knockdown of miR-483-5p or overexpression of miR-483-5p in EOC cell lines. Drug sensitivity assay was carried out by CCK-8 kit.Results:miR-483-5p was upregulated in EOC tissues as compared with normal tissues (P<0.01). Furthermore, miR-483-5p expression in advanced stage (Ⅲ–Ⅳ) EOC was significantly higher than that in early stage (Ⅰ–Ⅱ) EOC (P<0.05). Interestingly, miR-483-5p expression was higher in cisplatin-resistant A2780/CP cells than other cells. Increased miR-483-5p expression caused EOC cell resistance to cisplatin and downregulated the expression of p21 and Bcl-2, whereas reduced miR-483-5p expression induced its sensitivity and upregulated the expression of p21 and Bcl-2.Conclusion:The results suggest that miR-483-5p is highly expressed in EOC and contributes to cisplatin resistance. Thus, miR-483-5p is a potential therapeutic target for ovarian cancer.
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The objective of this study was to determine the expression of miR-483 and miR-483* and the relationship among them, their host gene (Igf2), and other cytokines in a murine model of renal fibrosis. The extent of renal fibrosis was visualized using Masson staining, and fibrosis was scored 3 days and 1 and 2 weeks after unilateral ureteral obstruction (UUO). Expression of miR-483, miR-483* and various cytokine mRNAs was detected by real-time polymerase chain reaction (PCR). Expression of miR-483 and miR-483* was significantly upregulated in the UUO model, particularly miR-483 expression was the greatest 2 weeks after surgery. Additionally, miR-483 and miR-483* expression negatively correlated with Bmp7 expression and positively correlated with Igf2, Tgfβ, Hgf, and Ctgf expression, as determined by Pearson's correlation analysis. Hgf expression significantly increased at 1 and 2 weeks after the surgery compared to the control group. This study showed that miR-483 and miR-483* expression was upregulated in a murine UUO model. These data suggest that miR-483 and miR-483* play a role in renal fibrosis and that miR-483* may interact with miR-483 in renal fibrosis. Thus, these miRNAs may play a role in the pathogenesis of renal fibrosis and coexpression of their host gene Igf2.
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Animais , Masculino , Camundongos , Expressão Gênica , Íntrons , Fator de Crescimento Insulin-Like II/genética , MicroRNAs , Obstrução Ureteral/genética , Obstrução Ureteral/patologia , Western Blotting , Citocinas/genética , Modelos Animais de Doenças , Fibrose/genética , Rim/patologia , Reação em Cadeia da Polimerase em Tempo Real , Fatores de TempoRESUMO
Objective To demonstrate that miR-483-5p and its targeted gene ERK1 involved in the regulation of proliferation and apoptosis of human granulosa cells.Methods ① The ERK1 mRNA and protein level expression were detected by PCR and Western blot after miR-483-5p overexpression in vitro normal human granular cells.De-tected the relative luciferase density after cotransfecting miR-483-5p mimics and its control respectively with wild or mutant ERK1 mRNA 3`UTR cloned luciferase report vector in granular cells.② Granular cells proliferation and ap-optosis were detected by MTT and TUNEL after transfecting miR-483-5p mimics or ERK1 siRNAs alone or simulta-neously.Results ① It showed that both ERK1 mRNA and protein in granular cells were markedly downregulated after the transfection of miR-483-5p mimics.A significant relative luciferase activity decrease were detected in the granular cells co-transfected with ERK1 wild and miR-483-5p mimics comparison with the control mimics,but not in the cells co-transfected with the ERK1 mutant and miR-483-5p mimics.② When miR-483-5p mimics and ERK1 siRNAs were alone or co-transfected into granular cells,the proliferation was inhibited while apoptosis trend was monitored to be promoted in all cases.No obvious statistic difference was shown between each other.Conclusion MiR-483-5p is involved in the regulation of the proliferation and apoptosis of human granulosa cells by directly binding to the target gene ERK1.