ABSTRACT
Objective To investigate the effect of mircoRNA-128-3p (miR-128-3p) on epithelial-mesencfrymal transition (EMT) of ovarian cancer cells and its regulatory mechanism on zinc finger E-box binding homobox 1(ZEB1). Methods Real-time PCR technology was used to detect the expression of miR-128-3p in epithelial ovarian cancer tissue (EOC) and adjacent normal tissue (30 cases each), and to observe whether there was a difference. Two human ovarian cancer cell lines, SK0V3 and A2780, were selected and transfected respectively. MiR-128-3p mimic (miR-128-3p mimic group) and negative control mimic (NC mimic group) were used to detect the expression of miR-128-3p in 4 groups by Real-time PCR to verify the interference effect. Transwell assay was used. The migration and invasion abilities of the four groups of cells were observed. The regulatory relationship between miR-128-3p and ZEBl was verified by dual luciferase assay, and the expression level of ZEBl protein after overexpression of miR-128-3p was detected by Western blotting; pcDNA-ZEBl was transfected into SK0V3 and A2780 cell lines to make it overexpression of ZEBl was divided into NC mimic group, miR-128-3p mimic group, and miR-128-3p mimic+pcDNA-ZEBl group. Western blotting was used to detect the EMT-related protein E-cadherin in 6 groups of cells and the expression level of vimentin. Results Real-time PCR result showed that the expression of miR-128-3p in EOC tissues decreased compared with that in adjacent tissues (P < 0. 01); The relative expression of miR-128-3p in the miR-128-3p mimic group was higher than that in the NC mimic group, while the numbers of migrating cells and invasive cells were lower than those in the NC mimic group (P < 0 . 0 1) . The result of dual luciferase experiments showed that miR-128-3p had a negative regulatory effect on ZEBl. In SK0V3 and A2780 cells, the relative expression of ZEBl protein in the miR-128-3p mimic group was lower than that in the NC mimic group, while the relative protein expression of E-cadhein was higher than that in the NC mimic group (P < 0 . 0 1) . The relative protein expression of E-cadhein in the miR-128-3p mimic+pcDNA-ZEBl group was lower than that in the miR-128-3p mimic group (P < 0 . 0 1) . In SKOV3 and A2780 cells, the relative protein expression of vimentin in the miR-128-3p mimic group was lower than that in the NC mimic group, and the relative protein expression of vimentin in the miR-128-3p mimic+pcDNA-ZEBl group was higher than that in the miR-128-3p mimic group (P < 0 . 0 1) . Conclusion The expression of miR-128-3p decreases in epithelial ovarian cancer tissues, which ma)' be due to the regulation of ZEBl to affect the expression of EMT-related proteins and participate in the EMT process of ovarian cancer cells.
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Objective:To detect the expression and clinical significance of microRNA-128-3p (miR-128-3p) in the plasma of patients with severe pancreatitis (SAP) .Methods:A total of 175 patients with acute pancreatitis who were treated in our hospital from Jun. 2017 to Dec. 2020 were selected as observation objects. According to the severity of the patients, the patients were divided into 78 cases in mild acute pancreatitis (MAP) group, 50 cases in moderate to severe acute pancreatitis (MSAP) group and 47 cases in severe acute pancreatitis (SAP) group; according to the patient’s prognosis, the SAP group was divided into 28 cases in the survival group and 19 cases in the death group. The level of miR-128-3p was detected by qRT-PCR method, and patients were evaluated with Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score and Ranson score, the correlation between plasma miR-128-3p level and APACHEⅡ, Ranson score was analyzed by Pearson, the predictive value of plasma miR-128-3p for poor prognosis of SAP patients was analyzed by ROC curve.Results:There were statistically significant differences in APACHE II[ (3.41±1.56) , (5.63±1.78) , (6.57±1.83) points], Ranson [ (2.58±1.34) , (4.95±1.47) , (6.06±1.92) points] and plasma CRP [ (39.73) ±12.31) , (70.25± 24.38) , (142.51±40.55) mg/L], serum calcium [ (1.95±0.31) , (13.26±5.24) , (14.21±6.32) mmol/L], among the MAP group, MSAP group and SAP group ( P<0.05) ; Compared with the MAP group (1.05±0.12) , the plasma miR-128-3p expression levels in the MSAP group (0.83±0.08) and the SAP group (0.57±0.05) were significantly decreased ( P<0.05) ; compared with the MSAP group (0.83±0.08) Compared with the SAP group (0.57±0.05) , the plasma miR-128-3p expression level was significantly decreased ( P<0.05) ; Compared with the survival group [ (0.65±0.08) , (5.91±1.64) points, (5.45±1.25) points, (120.43±30.56) mg/L], the plasma miR-128-3p of SAP patients in the death group was (0.43±0.05) expression levels were significantly reduced, APACHE II [ (7.43±2.21) points], Ranson score [ (7.22±1.59) points] and plasma CRP level [ (171.52±42.38) mg/L] were significantly increased ( P<0.05) ; the results of correlation analysis showed that plasma miR-128-3p level was negatively correlated with APACHEⅡ and Ranson scores ( r=-0.531, -0.609, P<0.05) ; The diagnostic efficacy of plasma miR-128-3p in predicting poor prognosis of SAP patients is better than APACHEⅡ, Ranson score, and CRP. Conclusion:Plasma miR-128-3p level is elevated in patients with severe pancreatitis, which is of certain value for the diagnosis and prognostic evaluation of SAP.
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Objective To investigate the effect of microRNA-128 (miR-128) on the proliferation,apoptosis on the hepatoma cells.Methods The target genes of miR-128 were analyzed by using bioinformatics and dual luciferase reporter assay system.We overexpressed miR-128 in the MHCC97H cells and also infected the GP73 gene with virus in the cell.Cell proliferation was detected by cell counting kit and apoptosis was detected by TUNEL and apoptosis index was calculated.Thirty 6-8 weeks Balb/c nude mice were randomly divided into miR-128 overexpression group (n =15) and control group (n =15).Results The results of bioinformatics and dual luciferase reporter assay showed that GP73 was the target gene of miR-128.Compared with the negative control group,cell proliferation in miR-128 overexpression group was decreased significantly;compared with miR-128 + empty plasmid group,cell in miR-128 + GP73 group was significantly increased (P ＜ 0.05).Compared with the negative control group,the apoptosis index in miR-128 overexpression group was significantly increased (P ＜ 0.05).Compared with miR-128 + empty plasmid group,the apoptosis index of miR-128 + GP73 group was significantly increased (P ＜ 0.05).Tumor volume of the overexpression group was higher than that of the control group.5 weeks later,tumor volume of the miR-128 overexpression group (1 209 ± 108) mm3 was significantly higher than that of the control group (1 985 ± 298)mm3,the difference was statistically significant (P ＜ 0.05).Conclusion miR-128 can promote the proliferation and inhibit the apoptosis of bepatoma cells by regulating GP73,which provide a reference for the prognosis evaluation of hepatoma.
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PURPOSE: Alzheimer's disease (AD) is the sixth most common cause of death in the United States. MicroRNAs have been identified as vital players in neurodegenerative diseases, including AD. microRNA-128 (miR-128) has been shown to be dysregulated in AD. This study aimed to explore the roles and molecular mechanisms of miR-128 in AD progression. MATERIALS AND METHODS: Expression patterns of miR-128 and peroxisome proliferator-activated receptor gamma (PPAR-γ) messenger RNA in clinical samples and cells were measured using RT-qPCR assay. PPAR-γ protein levels were determined by Western blot assay. Cell viability was determined by MTT assay. Cell apoptotic rate was detected by flow cytometry via double-staining of Annexin V-FITC/PI. Caspase 3 and NF-κB activity was determined by a Caspase 3 Activity Assay Kit or NF-κB p65 Transcription Factor Assay Kit, respectively. Bioinformatics prediction and luciferase reporter assay were used to investigate interactions between miR-128 and PPAR-γ 3′UTR. RESULTS: MiR-128 expression was upregulated and PPAR-γ expression was downregulated in plasma from AD patients and amyloid-β (Aβ)-treated primary mouse cortical neurons (MCN) and Neuro2a (N2a) cells. Inhibition of miR-128 decreased Aβ-mediated cytotoxicity through inactivation of NF-κB in MCN and N2a cells. Moreover, PPAR-γ was a target of miR-128. PPAR-γ upregulation attenuated Aβ-mediated cytotoxicity by inactivating NF-κB in MCN and N2a cells. Furthermore, PPAR-γ downregulation was able to abolish the effect of anti-miR-128 on cytotoxicity and NF-κB activity in MCN and N2a cells. CONCLUSION: MiR-128 inhibitor decreased Aβ-mediated cytotoxicity by upregulating PPAR-γ via inactivation of NF-κB in MCN and N2a cells, providing a new potential target in AD treatment.