الملخص
@#[Abstract] Objective: To investigate the regulating effects of miR-92b on the expression of EZH2 (enhancer of zeste homolog 2) gene and the proliferation and invasion abilities of esophageal cancer (EC) cells. Methods: Fifteen cases of esophageal cancer tissues that preserved in the research center of the Fourth HospitalAffiliated to Heibei Medical University from January 2016 to January 2017 were selected for this study. The bioinformatics tool was used to predict the possible miRNAs that might target EZH2. The mimics of predicted miRNAs were transfected into human esophageal carcinoma cell lines Eca109, respectively. Then the regulation effect of miRNAs on EZH2 gene expression was validated by real-time PCR, Western blotting and dual luciferase reporter experiment. In the meanwhile, EZH2 over-expression plasmids were co-transfected into esophageal carcinoma Eca109 cells, and the effects of miRNAs and EZH2 expression changes on the proliferation, apoptosis , invasion and migration of esophageal carcinoma cells were detected by CCK-8 method, Flow Cytometry, Transwell Invasion and migration assay, respectively. Results: Bioinformatics analysis showed that miR-92b, let7a and miR-25 could combine with potential binding sites at 3’-terminal non-translation region of EZH2 gene. Real-time PCR results showed that only miR-92b was able to regulate the expression of EZH2, and miR-92b was negatively correlated to EZH2 in esophageal cancer (P<0.01). Compared with mimic-NC, the expression of EZH2 mRNA, protein and luciferase activity in Eca109 cells after miR-92b mimic transfection was significantly down-regulated (both P<0.01). However, miR-92b mimic transfection had no effect on the apoptosis of Eca109 cells. Moreover, the proliferation, invasion and migration of Eca109 cells were significantly inhibited after transfection with miR-92b-mimic (P<0.01). In addition, after co-transfection with EZH2 over-expression plasmids, the effects of miR-92b-mimic on the proliferation, invasion and migration of Eca109 cells were significantly weakened (P<0.01). Conclusion: miR-92b can inhibit the proliferation,invasionandmigrationofesophagealcarcinomacells,anditsmechanismmayberelatedtoitstargetregulationofEZH2. ··
الملخص
[Objective]To investigate the role and the potential target of miR-92b-3p in angiotensin Ⅱ(Ang-Ⅱ)-induced mouse cardiac hypertrophy.[Methods]Ang-Ⅱ-induced cardiac hypertrophy models were established in adult C57BL/6 mice. AgomiR-92b-3p,the cholesterol-modified miR-92b-3p mimic,was delivered to increase the level of miR-92b-3p in mouse myocar?dium via tail vein injection. In the present study,three groups of mice were used in the animal experiment as follows,the agomiR-negative control(agomiR-NC)+saline group,the agomiR-NC+Ang-Ⅱgroup and the agomiR-92b-3p+Ang-Ⅱgroup. A cell model of cardiac hypertrophy was also established in Ang-Ⅱ-induced neonatal mouse cardiomyocytes in this study Luciferase activity was assayed after transfection using a luciferase reporter assay system. The expression of Myocyte-specific enhancer factor 2D( MEF2D) and hypertrophy-related genes atrial natriuretic peptide (ANP),cardiac muscle α-actin (ACTA1) and β-myosin heavy chain (MHC)at mRNA and protein levels in Ang-Ⅱ-induced hypertrophic myocardium and cardiomyocytes were detected by qRT-PCR and Western blot,respectively.[Results]The expression of ANP,ACTA1,β-MHC were markedly increased in Ang-Ⅱ-induced hypertrophic myocardium and cardiomyocytes. Dual luciferase reporter assay revealed that MEF2D is a potential target gene of miR-92b-3p. And miR-92b-3p can reduce the expression of MEF2D at the post-transcriptional level. Functionally,miR-92b-3p mimic, consistent with MEF2D siRNA,inhibited cell size increase and protein expression of ANP,ACTA1 andβ-MHC in Ang-II-treated mouse cardiomyocytes.[Conclusions]MEF2D is a novel target of miR-92b-3p,a target gene of miR-92b-3,which mediates the ef?fect of miR-92b-3p on attenuating cardiomyocyte hypertrophy.
الملخص
Objective To investigate the molecular mechanism of bone marrow mesenchymal stem cells (BM-MSCs) in repairing cisplatin-induced acute renal injury.Methods The rats were injected 6 mg/kg of cisplatin intraperitoneally,and bone marrow mesenchymal stem cells (BM-MSCs group) or PBS (PBS group) were injected respectively via tail vein after 24 hours.The rats without injecting cisplatin were selected as a normal control group.The repair effect of BM-MSCs on renal injury was observed by HE staining and immunohistochemistry.In addition,NRK-52E cells were cultured in vitro and treated with cisplatin for 6 hours.Then,NRK-52E cells were continued to culture for 48 hours or co-cultured with BM-MSCs for 48 hours,and NRK-52E cells untreated with cisplatin were used as a control.The expression levels of miR-92b and its target gene PTEN were detected by qRT-PCR,and the expression level of p-Akt by western blot.Results HE staining showed that the tubular protein casts in BM-MSCs group were significantly less than that in PBS group,and that the renal tubular structure was significantly improved in BM-MSCs group.Immunohistochemical staining indicated that the number of cells expressing proliferating cell nuclear antigen (PCNA) in BM-MSCs group (131.0 ± 14.4) was significantly higher than that in PBS group (42.2 ±6.1,t =11.28,P ＜0.01).qRT-PCR results showed that in the vivo experiment,compared with the expression level of miR-92b and PTEN in the normal control group (1.11 ± 0.78,1.01 ± 0.21),PBS group were (4.64 ± 1.06) and (0.61 ± 0.2),respectively (all P ＜ 0.05);BM-MSCs group were (2.27 ± 0.81) and (1.1 ± 0.1),respectively (all P ＜ 0.05).In vitro experiment,compared with the expression level of miR-92b and PTEN in the negative control group (1.12 ± 0.77,1.02 ± 0.13),cisplatin group were (7.64 ± 0.72) and (0.58 ± 0.2),respectively (all P ＜ 0.05),cell group were (4.38 ± 0.50) and (1.15 ± 0.23),respectively (all P ＜ 0.05).Western blot results showed that compared with the expression level of p-Akt in cisplatin group (0.96 ± 0.18),p-Akt expression in cell group was (2.11 ± 0.11,P ＜ 0.01).Conclusion BM-MSCs may repair the cisplatin-induced acute renal injury via down-regulating the expression level of miR-92b.
الملخص
Objective To compare the expression level of miR-92b in SHanG-44 glioma cells and glioma stem cells .Methods The tumor stem cells were screened from the SHG-44 glioma cells and identified by immunofluorescence .Then it was induced to differentiate .The transcription level of miR-92b was determined by RT-PCR and real-time quantitative PCR before and after differ-entiation .Results The miR-92b expression level in differentiated tumor cells was about 3 times higher than in tumor stem cells . Conclusion T he miR-92b gene is involved in the process of differentiation of glioma stem cells .