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@#AIM: To detect the relative expression levels of microRNA-27a(miR-27a)and nuclear factor erythroid-2-related factor 2(NRF2)in serum of patients with age-related macular degeneration(ARMD)bleeding, and to explore the correlation between the expression levels and the prognosis of ARMD bleeding. <p>METHODS: A retrospective case series observation was carried out.From June 2018 to October 2019, 80 patients with ARMD bleeding who were treated in our hospital were selected as ARMD bleeding group, and 80 healthy people who had routine examination in our hospital were selected as control group. The relative expression levels of miR-27a and NRF2 were detected by real-time fluorescent quantitative PCR(qRT-PCR), the diagnostic value of serum miR-27a and NRF2 expression for ARMD bleeding was evaluated by receiver operating characteristic curve(ROC). The incidence of poor prognosis was analyzed; in addition, Logistic regression was used to analyze the influencing factors of poor prognosis in patients with ARMD bleeding. <p>RESULTS: The relative expression level of miR-27a in serum of ARMD bleeding group was significantly higher than that of control group(<i>P</i><0.01), and the relative expression level of NRF2 mRNA in serum was significantly lower than that in control group(<i>P</i><0.01). ROC results showed that the AUC of serum miR-27a and NRF2 in the diagnosis of ARMD bleeding was 0.867 and 0.820 respectively, and the cutoff value was 1.10 and 1.08 respectively, at this time, the corresponding sensitivity was 71.3% and 91.3%, and the specificity was 90.0% and 63.7%, respectively. The AUC of serum miR-27a combined with NRF2 in the diagnosis of ARMD bleeding was 0.912, and the corresponding sensitivity and specificity were 86.3% and 85.0%, respectively. The incidence of poor prognosis in high miR-27a group was significantly higher than that in low miR-27a group(<i>P</i><0.05); and the incidence of poor prognosis in high NRF2 group was significantly lower than that in low NRF2 group(<i>P</i><0.05). Logistic analysis showed that the high expression of serum miR-27a was an independent risk factor for poor prognosis in patients with ARMD bleeding, and the high relative expression of NRF2 in serum was a protective factor for the poor prognosis of patients with ARMD bleeding. <p>CONCLUSION: The relative expression level of miR-27a in the serum of patients with ARMD hemorrhage is significantly increased, and the relative expression level of NRF2 is significantly decreased, both of them have certain diagnostic value for ARMD bleeding, and their relative expressions are closely related to the prognosis of patients, which is suggested that miR-27a and NRF2 can be used as potential biological indexes for early diagnosis and prognosis evaluation of ARMD bleeding.
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Aim To explore the role of miR-27a on the proliferation and metastasis of renal cell carcinoma (RCC) and its mechanism. Methods The expression of miR-27a in RCC cancer tissues, para-carcinoma tissues, RCC cells (Caki-1, 786-0 and ACHN) and normal renal tubular epithelial cells ( HK2) were detected by RT-qPCR. After miR-27a-inhibitor transfect-ed into 786-0 and ACHN cells, cell proliferation was measured by CCK-8 assay; cell colony formation was detected by colony formation assay; cell migration and invasion were detected by cell wound healing assay and Transwell assay, respectively; the pretein expression of P-catenin was detected by Western blot. After trans-fected miR-27a inhibitor into ACHN cells then treated with LiCl (Wnt/p-catenin signal agonist), cell proliferation , migration and invasion were detected. Results The expression of miR-27a in RCC cancer tissues was significantly higher than that in para-carcinoma tissues, and increased with stage progression. Compared with HK-2 cells, the expression levels of miR-27a in RCC cells were elevated. After transfection with miR-27a inhibitor, the cell colony formation, cell prolifera-tion, invasion and migration ability of 786-0 and ACHN cells were significantly reduced. MiR-27a in-i hibitor reduced the expression of p-catenin in ACHN cells. LiCl promoted the proliferation, invasion and migration ability of ACHN cells transfected with miR-27a inhibitor. Conclusions MiR-27a is highly expressed in RCC cancer tissues and RCC cells, and knockdown of miR-27a inhibits proliferation and metastasis in RCC cells through Wnt/p-catenin signaling pathway.
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Cysteine-rich secretory protein 2 (CRISP2) is an important protein in spermatozoa that plays roles in modulating sperm flagellar motility, the acrosome reaction, and gamete fusion. Spermatozoa lacking CRISP2 exhibit low sperm motility and abnormal morphology. However, the molecular mechanisms underlying the reduction of CRISP2 in asthenoteratozoospermia (ATZ) remain unknown. In this study, low expression of CRISP2 protein rather than its mRNA was observed in the ejaculated spermatozoa from ATZ patients as compared with normozoospermic males. Subsequently, bioinformatic prediction, luciferase reporter assays, and microRNA-27a (MIR-27a) transfection experiments revealed that MIR-27a specifically targets CRISP2 by binding to its 3' untranslated region (3'-UTR), suppressing CRISP2 expression posttranscriptionally. Further evidence was provided by the clinical observation of high MIR-27a expression in ejaculated spermatozoa from ATZ patients and a negative correlation between MIR-27a expression and CRISP2 protein expression. Finally, a retrospective follow-up study supported that both high MIR-27a expression and low CRISP2 protein expression were associated with low progressive sperm motility, abnormal morphology, and infertility. This study demonstrates a novel mechanism responsible for reduced CRISP2 expression in ATZ, which may offer a potential therapeutic target for treating male infertility, or for male contraception.
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Objective To discuss the effect of microRNA -27a on U251 glioma cells.Methods Over-expression or inhibition of miR -27a in U251 glioma cells were done by transient transfection of miR -27a mim-ics or AMO-27a in vitro.Cell viability was detected by MTT assay .Invasion ability of U251 was detected by tr-answell invasion assay.The level of miR-27a and PPARγwere detected by real -time PCR.Results Under in-hibition of miR-27a condition,the proliferation and invasiveness of U 251 glioma cells were decreased .The level of PPARγwas significantly increased ,whereas the level of miR-27 a was decreased .Overexpression of miR-27 a increased the proliferation and invasiveness of U 251 glioma cells and decreased the level of PPARγ.Conclusion Inhibition of miR-27a is benefit for inhibiting the proliferation and invasiveness of U 251 glioma cells.
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Objective:To explore the effects of miR-27a on the phenotype and cytokine secretion in LPS-stimulated dendritic cells.Methods:Murine bone marrow-derived dendritic cells were transfected with miR-27a mimics and negative control mimics,and then stimulated by LPS for 24 hours.Dendritic cells exposed to LPS were collected for analysis of the DC immunophenotype by flow cy-tometry and supernatants were collected to determine cytokine lever.Moreover,the capability of stimulating allogeneic CD4 T+cell prolif -eration was measured by MLR ( mixed lymphocyte reaction) .Results: The levels of MHCⅡ, CD80, and CD86 were significantly increased in LPS-stimulated dendritic cells when compared with imDC ( P<0.001).Transfection with miR-27a mimics resulted in sig-nificantly lower expression levels in levels of MHCⅡ,CD80,and CD86 (P<0.001).For cytokine secretion,transfection with miR-27a mimics enhanced IL-10 production (P<0.01) and reduced the production of IL-12 (P<0.05).For MLR,transfection with miR-27a mimics suppressed allogeneic CD4+T cell proliferation.Conclusion: MiR-27a may play critical roles in regulating the maturation process and cytokine secretion in LPS-stimulated dendritic cells.
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Objective To investigate the effect of miR-27a mimic and inhibitor on proliferation and apoptosis in melanoma cell line WM239. Methods The miR-27a mimic,inhibitor and its negative control were transfected into WM239 cells. The transfection efficiency was evaluated by fluorescence microscope. The expres-sion of miR-27a was detected by real-time fluorescent quantitative PCR. The proliferation of cells was detected by MTT. The cell apoptosis and cell cycle were analyzed by flow cytometry. Results The transfection efficiency in WM239 cells was 80% to 90%. The expression of miR-27a was markedly up-regulated in miR-27a mimic group (2-△△CT value is 26. 98 ± 0. 01),with statistically significant difference(t= -1 123. 67,P=0. 00);and the miR-27a inhibitor group showed lower expression of miR-27a(2-△△CT value is 0. 96 ± 0. 02),there was no statisti-cally significant difference compared with normal control group(t=0. 04,P=0. 06). The proliferation of cells was obviously inhibited in miR-27a mimic group,and there was statistically significant difference compared with normal control group[absorbance of 72 h(0. 45 ± 0. 02)∶(0. 72 ± 0. 01),F=129. 56,P﹤0. 05]. The percent-age of WM239 cells in G0-G1 phase was increased[(74. 83 ± 1. 46)∶(63. 73 ± 1. 25),F=30. 33,P﹤0. 05], and the percentage of WM239 cells in S phase and G2-M phase were decreased[(21. 33 ± 1. 75)∶(27. 50 ± 1. 25),F=14. 98,P﹤0. 05;(3. 90 ± 1. 31)∶(8. 80 ± 2. 10),F=3. 66,P﹤0. 05]. The apoptosis rate of cells was significantly increased in miR-27a mimic group compared with normal group[(29. 67 ± 0. 91)%∶(1. 44 ± 0. 85)%,F=530. 90,P﹤0. 01],but the inhibitor group had no obvious effect on cell cycle and cell apoptosis. Conclusion MiR-27a can suppress melanoma cell proliferation and act as a tumor suppressor gene,which is rel-evant to induce cell apoptosis and block cell cycle in G0-G1 phase.