Search

The paper aimed to explore the mechanism by which high mobility group box-1 and nicotinamide adenine dinucleotide phosphate oxidase-derived reactive oxygen species enhance each other's influence on the amplitude of electroretinogram and the permeability of the vitreous. According to the purpose of the experiment, the experimental rats were separated into a control group, a diabetes group and a Shenling Baizhu powder group. The messenger ribonucleic acid expression of microRNA-23 and microRNA-27a in rat muscle tissue was quantitatively analyzed by real-time quantitative polymerase chain reaction. The contractility of rat soleus skeletal muscle was analyzed by electrical stimulation program. Western blot analysis of rat muscle growth protein synthesis and the expression of myostatin signaling pathway was carried out. The expression of renal fibrosis-related albumin was tested by Western blot. The messenger ribonucleic acid expression of microRNA-23, 27a in the diabetic group was lower than control group (p<0.05), and the expression in the Shenling Baizhu powder group was higher than diabetic group (p<0.05). Compared with the control group, the diabetic group had lower body weight (p<0.05), and raised blood glucose concentration and diabetes volume (p<0.05). The ratio of phosphorylated protein kinase B/protein kinase B and phospho forkhead box protein O1/forkhead box protein O1 protein expression in the Shenling Baizhu powder group was higher than diabetic group (p<0.05), while the phosphatase and tensin homolog protein expression was lower (p<0.05). The protein expression of myostatin, FBXO32 and TRIM63 in the diabetes group was higher than control group (p<0.05), and the expression in the Shenling Baizhu powder group was lower than diabetes group (p<0.05). Compared with the control group, the protein expression of transforming growth factor-beta, suppressor of mothers against decapentaplegic 2/3, alpha-smooth muscle actin, collagen-1a and fibronectin was higher in the diabetes group (p<0.05), and the expression in the Shenling Baizhu powder group was lower than diabetes group (p<0.05). Shenling Baizhu powder can improve diabetes-induced sarcopenia by up-regulating microRNA expression, activating the insulin-like growth factor 1/phosphoinositide 3-kinases/ protein kinase B signaling pathway and inhibiting the myostatin cascade. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression of MicroRNA-27a (miR-27a) in the placenta of gestational diabetes mellitus (GDM) rats and its effects on oxidative stress and inflammation in GDM rats, as well as to analyze the underlying mechanism. Methods SD normal pregnant rats were injected intraperitoneally with strepto-zotocin (STZ) to induce the establishment of GDM models. After successful modeling, they were divided into four groups: model group, agomir-NC group, miR-27a agomir group, and miR-27a agomir + lipopolysaccharide (LPS) group, each with 12 rats; the control group consisted of12 normal pregnant rats. The levels of fasting blood glucose (FBG) and fasting insulin (FINS) in rats were detected, and the insulin resistance index (HOMA-IR) was calculated; the pregnancy outcome and placenta quality were analyzed; the levels of interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α), the activities of superoxide dismutase (SOD) and catalase (CAT), and the content of malondialdehyde (MDA) in placenta tissue were determined; Hematoxylin-Eosin (HE) staining and TUNEL method were used to detect pancreatic and placental tissue damage and cell apoptosis; and real -time quantitative PCR (RT - qPCR) and Western blot were used to detect the levels of miR-27a, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nuclear factor-κB p65 (NF-κB p65) mRNAs and proteins in placental tissues. Results Overexpression of miR-27a significantly reduced FBG, FINS and HOMA-IR in GDM rats, accompanied by a reduction in pro-inflammatory cytokine levels and oxidative stress, improved placental and pancreatic tissue damage and apoptosis, and the pregnancy outcome, and inhibited the activation of the TLR4/MyD88/NF -κB signaling pathway in the placenta (P < 0.05). Conclusion The overexpression of miR -27a can reduce the placental inflammation, oxidative stress, and IRin GDM rats. Its mechanism could be related to the inhibition of the TLR4/MyD88/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Abstract Background Oxidative stress plays a critical role on the processes of sepsis, and several microRNAs have been identified that may regulate the occurrence of oxidative stress. However, the relation between oxidative stress-related microRNA 27a (miR-27a) and sepsis is unknown. The present study aimed to determine the value of circulating miR-27a for the diagnosis and prognosis of sepsis. Methods This retrospective study included 23 patients with sepsis and 25 without sepsis treated at the emergency intensive care unit (EICU) or our institution between January 2019 and January 2020. Levels of circulating miR-27a and levels of oxidative stress-related indicators were measured and compared between sepsis and non-sepsis patients. Receiver operating characteristic (ROC) curve analysis was used to determine diagnostic efficiency of miR-27a. Results Circulating miR-27a levels in sepsis patients were higher than those in non-sepsis patients (p

The role of exosome-mediated mitophagy in the crosstalk between hepatocytes (HCs) and hepatic stellate cells (HSCs) in metabolic-associated fatty liver disease (MAFLD) remains unknown. Serum exosomal miR-27a levels were markedly increased and positively correlated with liver fibrosis in MAFLD patients and mice. Exosomal miR-27a was released from lipotoxic HCs and specifically transmitted to recipient-activated HSCs. PINK1, the key target of miR-27a, primarily mediates mitophagy. Overexpression of miR-27a or knockdown of PINK1 or lipotoxic HC-exosomal miR-27a impaired mitochondria (inhibiting mitophagy, respiration, membrane potential, and transcription while promoting reactive oxygen species production) in activated HSCs and stimulated HSC-derived fibroblasts (promoting activation and proliferation while inhibiting autophagy). High exosomal miR-27a serum levels and a lack of hepatic PINK1-mediated mitophagy were directly related to liver fibrosis in MAFLD mice. Lipotoxic HC exosome transplantation aggravated the degree of PINK1-mediated mitophagy suppression, steatohepatitis, lipidosis, and fibrosis in the livers of MAFLD mice with cirrhosis. Both in vitro and in vivo, exosomes derived from miR-27a-knockdown HCs could not facilitate the abovementioned deteriorating effects. In conclusion, lipotoxic HC-exosomal miR-27a plays a pivotal role in inhibiting mitophagy and in promoting MAFLD-related liver fibrosis by negatively regulating PINK1 expression.

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. 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. RESULTS: The relative expression level of miR-27a in serum of ARMD bleeding group was significantly higher than that of control group(P

Exosomes derived from cancer cells have emerged as important mediators of malignant phenotypes of tumors, being involved in the transmission of biological signals between cells. Herein, we intended to clarify the role of exosome-mediated transfer of oncogenic microRNA-27a (miR-27a) in angiogenesis of renal clear cell carcinoma (RCCC). Through bioinformatics analysis, we identified the differentially expressed genes of RCCC and predicted miRNAs targeting SFRP1. We manipulated the expression of miR-27a and/or SFRP1 in RCCC cells to explore their roles in angiogenesis through Cell Counting Kit-8 (CCK-8), Transwell, and Matrigel tubule formation assays. miR-27a loaded in exosomes was overexpressed and downregulated in vitro and in vivo to verify its effect on angiogenesis. SFRP1 was poorly expressed and miR-27a was highly expressed in RCCC tissues, showing a negative correlation. Dual-luciferase assay verified that miR-27a targeted and downregulated SFRP1 expression. Notably, miR-27a enhanced angiogenesis by downregulating SFRP1 expression. miR-27a-loaded exosomes can be delivered from RCCC cells to human umbilical vein endothelial cells (HUVECs). In vitro and in vivo experiments substantiated that miR-27a-loaded exosomes from RCCC cells repressed SFRP1, augmenting the viability, migration, and angiogenesis of RCCC cells. Together, RCCC-derived miR-27a-loaded exosomes inhibit SFRP1 expression and accelerate tumor angiogenesis in RCCC.

Serum-derived extracellular vesicles (EVs) containing non-coding RNAs have been indicated to serve as diagnostic and prognostic biomarkers for laryngeal squamous cell carcinoma (LSCC), while their functional role remains to be explored. Here, we summarize the possible mechanism explaining the laryngeal carcinogenesis and the associated changes with the involvement of extracellular microRNA (miR)-27a from serum of LSCC patients. Serum-derived EVs from LSCC patients were found to increase the proliferative activity and decreased the apoptotic activity of LSCC cells. miRNA microarrays revealed that miR-27a expression was elevated after EV treatment. miR-27a expression was elevated in LSCC tissues and predicted a poor prognosis for patients. Downregulation of miR-27a inhibited the effect of EVs to reduce the activity of LSCC cells in vitro and to suppress tumor development in vivo. miR-27a targeted SMAD family member 4 (Smad4) to mediate the Wnt/β-catenin pathway, which was induced under the influence of EVs. Smad4 was downregulated in LSCC tissues, and simultaneous overexpression of miR-27a and Smad4 resulted in reduced cell activity and tumorigenicity. In conclusion, serum-derived EVs support the laryngeal carcinogenesis at least partially via transferring miR-27a. miR-27a targets Smad4 and is a biomarker to predict LSCC prognosis. [ABSTRACT FROM AUTHOR]

Osteoporosis (OP) is a systemic skeletal disease that promotes bone fragility and the risk of fractures. Recent studies have shown the relevance of microRNAs (miRNAs) in the development of OP. This study aimed to evaluate the possible mechanisms of action underlying miR-27a loaded by mesenchymal stem cell (MSC)–derived extracellular vesicles (MSC-EVs) in OP. Serum samples from OP patients and normal controls were collected for miRNA microarray analysis. The expression of filtered miRNA was upregulated in osteoblasts (OB) and osteoclasts (OCs) for biological activity assessment. After developing OP mice using ovariectomy (OVX) and confirming OP, the miR-27a expression level was upregulated in mice by MSC-EV application. Dual-luciferase assays were conducted to validate the relationship between miR-27a and DKK2 expression. The poor expression of miR-27a was observed in patients with OP. miR-27a increased the expression of OB markers, the number of ALP-positive cells, and the number of calcium nodules in OCs. In OVX mice, miR-27a increased bone density, improved bone structure damage recovery, decreased the levels of bone resorption markers, and decreased OC number. miR-27a transmitted by MSC-EVs interacted with DKK2. MSC-EVs exerted the same protective effects as miR-27a on OP, whereas miR-27a inhibitor abolished the attenuating effects of MSC-EVs. In contrast, DKK2 depletion reversed the stimulatory effects of the miR-27a inhibitor on OP. The Wnt/β-catenin pathway was activated upon MSC-EV application and DKK2 silencing and was impaired upon the downregulation of the expression of miR-27a. MSC-EVs are effective in preventing mouse OP. This mechanism is mediated by the miR-27a/DKK2/Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Oxidative stress plays a critical role on the processes of sepsis, and several microRNAs have been identified that may regulate the occurrence of oxidative stress. However, the relation between oxidative stress-related microRNA 27a (miR-27a) and sepsis is unknown. The present study aimed to determine the value of circulating miR-27a for the diagnosis and prognosis of sepsis. Methods: This retrospective study included 23 patients with sepsis and 25 without sepsis treated at the emergency intensive care unit (EICU) or our institution between January 2019 and January 2020. Levels of circulating miR-27a and levels of oxidative stress-related indicators were measured and compared between sepsis and non-sepsis patients. Receiver operating characteristic (ROC) curve analysis was used to determine diagnostic efficiency of miR-27a. Results: Circulating miR-27a levels in sepsis patients were higher than those in non-sepsis patients (p < 0.05), and levels were significantly higher in patients that died than those that lived (p < 0.05). In patients with sepsis, circulating miR-27a level was positively correlated with serum malondialdehyde (MDA) level (rs = 0.529, p = 0.007), and negatively correlated with serum glutathione peroxidase (GSH-Px) level (rs = − 0.477, p = 0.016). No significant correlation was observed between circulating miR-27a and serum superoxide dismutase (SOD) in sepsis patients (rs = − 0.340, p = 0.096). The area under the ROC curve (AUC) of miR-27a level for prediction of sepsis was 0.717 (p = 0.009) and for 28-day mortality was 0.739 (p = 0.003). Conclusions: This study showed that circulating miR-27a level is correlated with oxidative stress and mortality in patients with sepsis, and may serve as a potential non-invasive molecular biomarker. [ABSTRACT FROM AUTHOR]

Arbutin (ARB) has been widely used in skin pigmentation disorders. Nevertheless, the involvements of ARB in diabetic nephropathy (DN) are still unknown. We investigated the functions of ARB in high glucose (HG)-induced cell apoptosis and autophagy in HK-2 cells. Cell viability was examined through CCK-8 in HK-2 cells after disposal with 45 mM glucose and ARB (10-50 μM). Flow cytometry and western blot tested cell apoptosis and the related protein levels in HK-2 cells after 45 mM glucose and 50 μM ARB administration. RT-qPCR delved microRNA (miR)-27a expression in HG and ARB co-treated HK-2 cells. Effect of miR-27a on ARB affected cell apoptosis and autophagy was investigated after miR-27a inhibitor transfection. JNK and mTOR pathways were finally assessed by western blot. ARB alleviated HG-induced cell apoptosis, autophagy and regulated the related protein levels in HK-2 cells. MiR-27a expression was reduced in HG-treated cells, but was accelerated in HG and ARB co-treated HK-2 cells with the increased concentration. Inhibition of miR-27a apparently abolished the outcomes of ARB in HG-induced HK-2 cells apoptosis and autophagy. Besides, ARB blocked JNK and mTOR pathways by regulating miR-27a. The findings demonstrated that ARB alleviated apoptosis and autophagy in HG-treated HK-2 cells by regulating miR-27a/JNK/mTOR axis.

Salidroside (Sal) exerted widely pharmacological effects in multitudinous diseases had been certified. The actual study clarified the protective activity of Sal in H2O2-injured human trabecular meshwork (HTM) cells. HTM cells were disposed with H2O2 to construct an oxidative damage model in vitro. Then, Sal was utilized to administrate HTM cells, and cell viability, apoptosis, apoptosis-interrelated proteins and ROS production were appraised using CCK-8, flow cytometry, western blot and DCFH-DA staining. MiR-27a inhibitor and its control were transfected into HTM cells, and the influences of miR-27a inhibition in HTM cells stimulated with H2O2 and Sal were detected. PI3K/AKT and Wnt/β-catenin pathways were ultimately investigated to uncover the underlying mechanism. We found that H2O2 evoked HTM cells oxidative damage, as evidenced by repressing cell viability, inducing apoptosis, activating cleaved-caspase-3/-9 expression and increasing ROS production. Sal significantly lightened H2O2-evoked oxidative damage in HTM cells. Additionally, miR-27a was up-regulated by Sal, and miR-27a suppression significantly reversed the protective effect of Sal on H2O2-injured HTM cells. Finally, Sal activated PI3K/AKT and Wnt/β-catenin pathways through enhancement of miR-27a in H2O2-injured HTM cells. In conclusion, these discoveries suggested that Sal could protect HTM cells against H2O2-evoked oxidative damage by activating PI3K/AKT and Wnt/β-catenin pathways through enhancement of miR-27a.HighlightsH2O2 evokes HTM cells oxidative damage;Sal relieves H2O2-induced oxidative damage in HTM cells;Sal enhances miR-27a expression in H2O2-injured HTM cells;Repressed miR-27a reverses the protective impacts of Sal on H2O2-injured HTM cells;Sal activates PI3K/AKT and Wnt/β-catenin pathways by increasing miR-27a.

Pulmonary artery hypertension (PAH) is a common and serious disease which is characterized by pulmonary vascular remodeling. Bosentan (BST) is the first approved oral targeted drug of endothelin-1 (ET-1) receptor antagonists for the treatment of PAH. MicroRNA-27a (miR-27a) and peroxisome proliferator-activated receptor γ (PPARγ) were found to be related to the pathogenesis of PAH. To further explore the signal transduction mechanism of BST in the treatment of PAH, we examined the effects of BST on endothelin receptors, miR-27a, and PPARγ. Meanwhile, the influence of miR-27a in the formation and development of PAH was discussed. Our results demonstrated that during the pathophysiology of PAH, miR-27a, PPARγ, and ET-1 were cross-inhibited, which indicated that the miR-27a/PPARγ/ET-1 signaling pathway was dysregulated; in addition, BST could competitively bind to ET-1 receptors and inhibit the miR-27a/PPARγ/ET-1 signaling pathway, thereby delaying the proliferation of PASMCs and affecting the development of PAH. Our results give a new understanding of the pathogenesis and treatment of PAH and provide more reliable evidence for the application of BST in the treatment of PAH in the clinic. [ABSTRACT FROM AUTHOR]

The function of microRNA-27a (miR-27a) expression in cholangiocarcinoma (CCA) remains largely unclear; therefore, this study aimed to investigate the clinical significance and functional role of miR-27a in CCA. This study included 117 paired CCA tissues and adjacent normal tissues from CCA patients who received surgical resection. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression levels of miR-27a in CCA tissues and cell lines. A Kaplan–Meier curve and Cox regression analysis were used to determine overall prognostic performance. The effects of miR-27a on cell proliferation, migration, and invasion were measured by CCK-8 and Transwell assays. The expression levels of miR-27a in patients with CCA and cell lines were higher than those in adjacent normal tissues and normal cells, respectively. Additionally, miR-27a levels were found to be associated with lymph node metastasis and TNM stages. The overall survival time of CCA patients with high miR-27a expression was poorer than that of those with low miR-27a expression. Furthermore, miR-27a overexpression promoted CCA cell proliferation, migration, and invasion, whereas knockdown of miR-27a suppressed cell proliferation, migration, and invasion. Taken together, these results suggest the potential usefulness of miR-27a in the prognosis and progression of CCA. [ABSTRACT FROM AUTHOR]

Objective: To investigate the angiogenesis of human microvascular endothelial cells (HMEC-1) induced by human nucleus pulposus cells (NPCs) through miRNA-27a / SPRY2. Methods: Intervertebral disc tissues from scoliosis and intervertebral disc degeneration (IDD) patients were collected as control group and IDD group, and differentially expressed miRNAs were screened by miRNA chip. The expression of miR-27a in tissues was verified by RT-qPCR and fluorescence in situ hybridization. Lentivirus-transfected cells were divided into Control group (un transfected); NC group (transfected with lv-pNanog); sh-miR-27a group (transfected with miR-27a knockdown plasmid); miR-27a group (transfected with miR-27a overexpression plasmid); SPRY2 group (transfected with SPRY2 overexpression plasmid) and miR-27a + SPRY2 group (transfected with miR-27a and SPRY2 overexpression plasmid). The expression of miR-27a and SPRY2 in nucleus pulposus cells was detected by RT-qPCR. verified the The targeting relationship between miR-27a and SPRY2 was verified by dual-luciferase reporter gene assay. HMEC-1 cells were cultured by mixed medium which is mixed by conditioned medium of nucleus pulposus and complete medium. Trans well and lumen formation experiments were used to detect the invasion and angiogenesis ability of HMEC-1 cells. Transforming growth factor-β1 (TGF-β1) expreesion in nucleus pulposus cells and mixed medium was detected by immunofluorescence and ELISA. Results: Compared with the control group, the expression of miR-27a in IDD group was significantly increased. Compared with the NC group, the expression of SPRY2 was increased in the sh-miR-27a group (P<0.05) and decreased in the miR-27a group (P <0.05). Compared with the NC group, the invasion and angiogenesis of HMEC-1 cells in the miR-27a group were enhanced (P<0.05), and the expression of TGF-β1 was increased (P<0. 05); Invasion of HMEC-1 cell in the SPRY2 group was reduced (P<0.05), the lumen-like structure was not formed. Compared with the miR-27a group, the invasion and angiogenesis ability of HMEC-1 cells in the miR-27a + SPRY2 group reduced (P<0.05), and the expression of TGF-β1 decreased (P<0.05). miRNA-27a promotes the ability of nucleus pulposus cells to induce angiogenesis of HMEC-1 cells by inhibiting the expression of SPRY2. [ABSTRACT FROM AUTHOR]

Curcumin (CUR) is a kind of polyphenolic compound and widely used in the treatment of diseases. However, the involvement of CUR in thymic carcinoma remains unknown. The object of our research is to clarify the role of CUR and related regulatory mechanism in thymic carcinoma cells. After treatment with CUR for 24 hr, cell viability, apoptosis, migration, and invasion of TC1889 cells were measured. Real-time polymerase chain reaction was executed to examine the expression of microRNA-27a (miR-27a) in thymic carcinoma tissues and TC1889 cells. After miR-27a mimic transfection, whether miR-27a is involved in CUR-modulated cell behaviors was measured. Finally, western blot was utilized to detect mTOR and Notch 1 pathways-linked proteins. CUR restrained cell viability and increased cell apoptosis of TC1889 cells. In addition, cell migration and invasion were restrained by CUR. Meanwhile, miR-27a expression was positively regulated in thymic carcinoma tissues and downregulated by CUR in TC1889 cells. Overexpressed miR-27a reversed the CUR-induced reduction of growth, migration, and invasion in TC1889 cells. Furthermore, CUR blocked mTOR and Notch 1 pathways via downregulating miR-27a. We demonstrated that CUR blocked mTOR and Notch 1 pathways via downregulating miR-27a, thereby suppressing cell growth, migration, and invasion of thymic carcinoma cells. [ABSTRACT FROM AUTHOR]

The aims of the present study were to examine the clinical significance of serum microRNA-27a (miR-27a) levels in patients with multiple organ dysfunction syndrome (MODS) caused by acute paraquat poisoning and to investigate the correlation between miR-27a and interleukin (IL)-10. A total of 82 patients with MODS induced by acute paraquat poisoning and 88 healthy controls were recruited in the present study. Reverse transcription-quantitative PCR was used to measure serum miR-27a expression levels in patients with MODS and the control group. IL-10 serum levels were determined using ELISA. Decreased serum miR-27a level and increased IL-10 expression levels were detected in patients with paraquat poisoning compared with healthy controls (P<0.001). A moderately negative correlation was identified between the serum expression levels of miR-27a and IL-10 (r=−0.5225; P<0.001). miR-27a expression level was found to be associated with blood urea nitrogen, partial pressure of carbon dioxide, arterial blood lactic acid, and the acute physiology and chronic health evaluation II score (APACHE II; P<0.05). The area under the curve for miR-27a was 0.946, with a sensitivity of 86.6% and specificity of 87.5% at a cutoff value of 2.10. The non-survival patient group had lower miR-27a expression levels compared with the survival group (P<0.01). Multivariate Cox regression analyses suggested miR-27a expression level and APACHE II score were independent prognostic factors for 30-day mortality (P<0.01). The present results suggested that serum miR-27a level may be a potential novel diagnostic and prognostic factor for MODS caused by paraquat poisoning. Collectively, miR-27a may be involved in the process of MODS induced by paraquat poisoning by regulating the inflammatory response. [ABSTRACT FROM AUTHOR]

Background/Aims: We examined the effects of microRNA-27a (miR-27a) on detrusor fibrosis in streptozotocin (STZ)-induced diabetic rats. Methods: Eighty healthy Sprague-Dawley (SD) rats were randomly allocated into control, diabetic, miR-27a mimics, mimics control, miR-27a inhibitors, inhibitors control, siRNA-PRKAA2 (siPRKAA2) and inhibitors + siPRKAA2 groups (the latter 7 groups were established as STZ-induced diabetic rat models and treated in different manners). Detrusor cell apoptosis in bladder tissues was determined through terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining. Detrusor cells were assigned to the blank, miR-27a mimics, mimics control, miR-27a inhibitors, inhibitors control, siPRKAA2 and inhibitors + siPRKAA2 groups. Flow cytometry determined the cell cycle stage and apoptosis. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting (WB) were used to assess the expression of miR-27a, PRKAA2, TGF-β1, Smad3, p-Smad3, fibronectin (FN), connective tissue growth (CTGF), and collagen-I (COL-I) in tissues and cells. Results: Compared with the control group, the diabetic, miR-27a mimics, and siPRKAA2 groups showed reduced weight and PRKAA2 expression, but elevated blood glucose, serum creatinine (sCr), blood urea nitrogen (BUN), cell apoptosis, and expression of TGF-β1, Smad3, FN, COL-I, CTGF, and p-Smad3. The opposite trend was observed in the miR-27a inhibitors group. PRKAA2 is a target gene of miR-27a. Compared to the blank group, the miR-27a mimics and siPRKAA2 groups indicated markedly increased TGF-β1, Smad3, FN, COL-I, CTGF and p-Smad3 expression; decreased PRKAA2 expression; and increased cell apoptosis. The miR-27a inhibitors group showed the opposite trend. Conclusion: These results indicate that miR-27a may contribute to detrusor fibrosis in STZ-induced diabetic rats by targeting PRKAA2 via the TGF-β1/Smad3 signaling pathway.

MicroRNAs (miRNAs), a class of small, non-coding RNA molecules, play an important role in the posttranscriptional regulation of gene expression, thereby influencing important cellular functions. In adipocytes, miRNAs show import regulatory features and are described to influence differentiation as well as metabolic, endocrine, and inflammatory functions. We previously identified miR-27a being upregulated under inflammatory conditions in human adipocytes and aimed to elucidate its function in adipocyte biology. Both strands of miR-27a, miR-27a-3p and -5p, were downregulated during the adipogenic differentiation of Simpson–Golabi–Behmel syndrome (SGBS) cells, human multipotent adipose-derived stem cells (hMADS), and human primary adipose-derived stromal cells (hASCs). Using miRNA-mimic transfection, we observed that miR-27a-3p is a crucial regulator of adipogenesis, while miR-27a-5p did not alter the differentiation capacity in SGBS cells. In silico screening predicted lipoprotein lipase (LPL) and peroxisome proliferator activated receptor γ (PPARγ) as potential targets of miR-27a-3p. The downregulation of both genes was verified in vitro, and the interaction of miR-27-3p with target sites in the 3′ UTRs of both genes was confirmed via a miRNA-reporter-gene assay. Here, the knockdown of LPL did not interfere with adipogenic differentiation, while PPARγ knockdown decreased adipogenesis significantly, suggesting that miR-27-3p exerts its inhibitory effect on adipogenesis by repressing PPARγ. Taken together, we identified and validated a crucial role for miR-27a-3p in human adipogenesis played by targeting the essential adipogenic transcription factor PPARγ. Though we confirmed LPL as an additional target of miR-27a-3p, it does not appear to be involved in regulating human adipogenesis. Thereby, our findings call the conclusions drawn from previous studies, which identified LPL as a crucial regulator for murine and human adipogenesis, into question.

Objective: This study aims to explore the effects of microRNA-27a (miR-27a) on the proliferation and invasiveness of colon cancer cells through the Secreted Frizzled-related protein 1 (SFRP1) and the Wnt/β-catenin signaling pathway. Methods: Colon cancer tissues and adjacent normal tissues from 125 colon cancer patients, together with the HCEpic, HCT-116, HT-29, SW480 and SW620 cell lines, were prepared for this study. The transfected HCT-116 cells were divided into the miR-27a mimics, miR-27a-NC, anti-miR-27a, blank, Lv-SFRP1, Lv-NC, and miR-27a mimics + Lv-SFRP1 groups. RT-qPCR was performed to detect the expressions of miR-27a and SFRP1 mRNA. A dual-luciferase reporter assay was conducted to examine the effect of miR-27a on SFRP1. Western blotting was used to measure the expressions of the SFRP1, β-catenin, GSK-3β, p-β-catenin, p-GSK-3β, c-Myc and cyclin D1 proteins. MTT, soft agar clone formation and Transwell chamber assays were performed to detect cell proliferation and invasion. Results: Compared with normal tissues and cells, colon cancer tissues and cells demonstrated significantly higher expression of miR-27a, but lower expressions of SFRP1 mRNA and protein. MiR-27a negatively regulated the expression of SFRP1 mRNA. SFRP1 was also found to be a target gene of miR-27a. In the miR-27a mimic group, the proliferation and invasiveness of colon cancer cells were significantly increased, while the expressions of GSK-3 β and p-β-catenin were remarkably down-regulated; in contrast, the expressions of p-GSK-3β, -catenin, c-Myc and cyclin D1 were up-regulated. While the proliferation and invasiveness of colon cancer cells in the anti-miR-27a and Lv-SFRP1 groups were decreased, the expressions of GSK-3β and p-β-catenin were elevated, and the expressions of p-GSK-3β, β-catenin, c-Myc and cyclin D1 were decreased. Conclusion: These findings indicated that miR-27a could promote the proliferation and invasiveness of colon cancer cells by targeting SFRP1 through the Wnt/β-catenin signaling pathway.

MicroRNAs (miRNAs) are a family of highly conserved, non-coding single-stranded RNAs transcribed as ~70 nucleotide precursors to an 18–22 nucleotide product (1). miRNAs can silence their homologous target genes at the post-transcriptional level, and these genes have been revealed to play an important role in tumorigenesis, invasion and metastasis (2). MicroRNA-27a (miR-27a), transcripted by miR-27a gene, has proved to implicate with many kinds of solid tumors, showing potential as a useful biomarker or drug target for clinical application. However, even though miR-27a has been reported in many cancers, the mechanism and signal pathways of miR-27 in oncogenesis, invasion, and metastasis are still obscure. Moreover, recent studies show that miR-27a pays an important role in epithelial-mesenchymal-transition, regulating tumor immune response, and chemoresistance. In this review, we summarize the current literature, demonstrate the established link between miR-27a and tumorigenesis, and focus on recently identified mechanisms. The review also aims to demonstrate the potential of miR-27a as a diagnostic and/or prognostic biomarker in solid tumors and to discuss the possibilities of targeted therapy and drug design.

Cardiac hypertrophy is the most prevalent compensatory heart disease that ultimately leads to spontaneous heart failure. Mounting evidence suggests that microRNAs (miRs) and endogenous hydrogen sulfide (H 2 S) play a crucial role in the regulation of cardiac hypertrophy. In this study, we aimed to investigate whether inhibition of miR-27a could protect against cardiac hypertrophy by modulating H 2 S signaling. We established a model of cardiac hypertrophy by obtaining hypertrophic tissue from mice subjected to transverse aortic constriction (TAC) and from cells treated with angiotensin-II. Molecular alterations in the myocardium were quantified using quantitative real time PCR (qRT-PCR), Western blotting, and ELISA. Morphological changes were characterized by hematoxylin and eosin (HE) staining and Masson's trichrome staining. Functional myocardial changes were assessed using echocardiography. Our results demonstrated that miR-27a levels were elevated, while H 2 S levels were reduced in TAC mice and myocardial hypertrophy. Further luciferase and target scan assays confirmed that cystathionine-γ-lyase (CSE) was a direct target of miR-27a and was negatively regulated by it. Notably, enhancement of H 2 S expression in the heart was observed in mice injected with recombinant adeno-associated virus vector 9 (rAAV9)-anti-miR-27a and in cells transfected with a miR-27a inhibitor during cardiac hypertrophy. However, this effect was abolished by co-transfection with CSE siRNA and the miR-27a inhibitor. Conversely, injecting rAAV9-miR-27a yielded opposite results. Interestingly, our findings demonstrated that glucagon-like peptide-1 (GLP-1) agonists could mitigate myocardial damage by down-regulating miR-27a and up-regulating CSE. In summary, our study suggests that inhibition of miR-27a holds therapeutic promise for the treatment of cardiac hypertrophy by increasing H 2 S levels. Furthermore, our findings unveil a novel mechanism of GLP-1 agonists involving the miR-27a/H 2 S pathway in the management of cardiac hypertrophy., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Pancreatic cancer (PC) remains a primary cause of cancer‐related deaths worldwide. Existing literature has highlighted the oncogenic role of microRNA‐27a (miR‐27a) in multiple cancers. Hence, the current study aimed to clarify the potential therapeutic role of PC cell–derived exosomal miR‐27a in human microvascular endothelial cell (HMVEC) angiogenesis in PC. Initially, differentially expressed genes (DEGs) and miRs related to PC were identified by microarray analysis. Microarray analysis provided data predicting the interaction between miR‐27a and BTG2 in PC, which was further verified by the elevation or depletion of miR‐27a. Next, the expression of miR‐27a and BTG2 in the PC tissues was quantified. HMVECs were exposed to exosomes derived from PC cell line PANC‐1 to investigate the effects associated with PC cell–derived exosomes carrying miR‐27a on HMVEC proliferation, invasion and angiogenesis. Finally, the effect of miR‐27a on tumorigenesis and microvessel density (MVD) was analysed after xenograft tumour inoculation in nude mice. Our results revealed that miR‐27a was highly expressed, while BTG2 was poorly expressed in both PC tissues and cell lines. miR‐27a targeted BTG2. Moreover, miR‐27a silencing inhibited PC cell proliferation and invasion, and promoted apoptosis through the elevation of BTG2. The in vitro assays revealed that PC cell–derived exosomes carrying miR‐27a stimulated HMVEC proliferation, invasion and angiogenesis, while this effect was reversed in the HMVECs cultured with medium containing GW4869‐treated PANC‐1 cells. Furthermore, in vivo experiment revealed that miR‐27a knockdown suppressed tumorigenesis and MVD. Taken together, cell‐derived exosomes carrying miR‐27a promotes HMVEC angiogenesis via BTG2 in PC. [ABSTRACT FROM AUTHOR]

The function of microRNA‐27a (miR‐27a) on synovial angiogenesis and chondrocyte apoptosis in rats with knee osteoarthritis (KOA) by targeting PLK2 is explored in this present study. The rat model of KOA was conducted by anterior cruciate ligament transection. Rats were injected with miR‐27a mimics, mimics NC, pcDNA3.1‐PLK2 pcDNA3.1, or RLK2 RNAi plasmid via tail vein. A series of assays were used to figure out the functions of miR‐27a and PLK2 in synovial angiogenesis and chondrocyte apoptosis in rats with KOA. Furthermore, the putative binding site between miR‐27a and PLK2 was determined. Downregulated miR‐27a was found in synovial tissues and cartilage tissues of KOA rats. Upregulated miR‐27a and downregulated PLK2 inhibited synovial injury and promoted apoptosis of synovial cells, inhibited synovial angiogenesis, inhibited cartilage injury and chondrocyte apoptosis, inhibited cartilage collagen destruction, and alleviates inflammatory injury of synovial tissue and cartilage tissue in KOA rats. Overexpression of PLK2 reverses the effect of upregulation of miR‐27a on synovial angiogenesis and chondrocyte injury in KOA rats. Our study suggests that upregulation of miR‐27a inhibits synovial angiogenesis and chondrocyte apoptosis in KOA rats through the inhibition of PLK2. [ABSTRACT FROM AUTHOR]

Renal tubular damage caused by persistent high glucose environment has been found to contribute to diabetic nephropathy. This study explored the effects of lncRNA growth arrest-specific 5 (GAS5) on high glucose-stimulated human renal tubular epithelial HK-2 damage, as well as the possible internal molecular mechanism. Viability and apoptosis of HK-2 cells were assessed with the help of CCK-8 assay and Annexin V-FITC/PI staining, respectively. Cell transfection was used to change the expression of GAS5, miR-27a and BNIP3. We found that high glucose stimulation suppressed HK-2 cell viability but induced cell apoptosis. The expression of GAS5 was increased in HK-2 cells under high glucose environment. Silence of GAS5 mitigated the high glucose-caused HK-2 cell viability reduction and apoptosis. Overexpression of miR-27a reversed the effects of GAS5 on high glucose-stimulated HK-2 cells. Overexpression of BNIP3 aggravated the high glucose-caused HK-2 cell viability reduction, apoptosis and activation of JNK pathway. Knockdown of BNIP3 had opposite effects. In conclusion, this research further confirmed the pro-apoptotic roles of GAS5 in renal tubular epithelial cells under high glucose environment. Silence of GAS5 alleviated high glucose toxicity to human renal tubular epithelial HK-2 cells might be via down-regulating miR-27a and BNIP3, and then inactivating JNK pathway. HG suppresses HK-2 cell viability, but promotes cell apoptosis; HG enhances the expression of GAS5 in HK-2 cells; Silence of GAS5 alleviates the HG-caused HK-2 cell toxicity; miR-27a participates in the effects of GAS5 silencing on HG-stimulated HK-2 cells; BNIP3 is regulated by miR-27a and related to the HG toxicity to HK-2 cells. [ABSTRACT FROM AUTHOR]

Arbutin (ARB) has been widely used in skin pigmentation disorders. Nevertheless, the involvements of ARB in diabetic nephropathy (DN) are still unknown. We investigated the functions of ARB in high glucose (HG)-induced cell apoptosis and autophagy in HK-2 cells. Cell viability was examined through CCK-8 in HK-2 cells after disposal with 45 mM glucose and ARB (10-50 μM). Flow cytometry and western blot tested cell apoptosis and the related protein levels in HK-2 cells after 45 mM glucose and 50 μM ARB administration. RT-qPCR delved microRNA (miR)-27a expression in HG and ARB co-treated HK-2 cells. Effect of miR-27a on ARB affected cell apoptosis and autophagy was investigated after miR-27a inhibitor transfection. JNK and mTOR pathways were finally assessed by western blot. ARB alleviated HG-induced cell apoptosis, autophagy and regulated the related protein levels in HK-2 cells. MiR-27a expression was reduced in HG-treated cells, but was accelerated in HG and ARB co-treated HK-2 cells with the increased concentration. Inhibition of miR-27a apparently abolished the outcomes of ARB in HG-induced HK-2 cells apoptosis and autophagy. Besides, ARB blocked JNK and mTOR pathways by regulating miR-27a. The findings demonstrated that ARB alleviated apoptosis and autophagy in HG-treated HK-2 cells by regulating miR-27a/JNK/mTOR axis. [ABSTRACT FROM AUTHOR]

Salidroside (Sal) exerted widely pharmacological effects in multitudinous diseases had been certified. The actual study clarified the protective activity of Sal in H2O2-injured human trabecular meshwork (HTM) cells. HTM cells were disposed with H2O2 to construct an oxidative damage model in vitro. Then, Sal was utilized to administrate HTM cells, and cell viability, apoptosis, apoptosis-interrelated proteins and ROS production were appraised using CCK-8, flow cytometry, western blot and DCFH-DA staining. MiR-27a inhibitor and its control were transfected into HTM cells, and the influences of miR-27a inhibition in HTM cells stimulated with H2O2 and Sal were detected. PI3K/AKT and Wnt/β-catenin pathways were ultimately investigated to uncover the underlying mechanism. We found that H2O2 evoked HTM cells oxidative damage, as evidenced by repressing cell viability, inducing apoptosis, activating cleaved-caspase-3/-9 expression and increasing ROS production. Sal significantly lightened H2O2-evoked oxidative damage in HTM cells. Additionally, miR-27a was up-regulated by Sal, and miR-27a suppression significantly reversed the protective effect of Sal on H2O2-injured HTM cells. Finally, Sal activated PI3K/AKT and Wnt/β-catenin pathways through enhancement of miR-27a in H2O2-injured HTM cells. In conclusion, these discoveries suggested that Sal could protect HTM cells against H2O2-evoked oxidative damage by activating PI3K/AKT and Wnt/β-catenin pathways through enhancement of miR-27a. H2O2 evokes HTM cells oxidative damage; Sal relieves H2O2-induced oxidative damage in HTM cells; Sal enhances miR-27a expression in H2O2-injured HTM cells; Repressed miR-27a reverses the protective impacts of Sal on H2O2-injured HTM cells; Sal activates PI3K/AKT and Wnt/β-catenin pathways by increasing miR-27a. [ABSTRACT FROM AUTHOR]

Liver ischemia-reperfusion (I/R) injury is an important clinical issue related to liver transplantation. Recent studies suggest that microRNAs are implicated in various biological and pathological processes, including liver I/R injury. This study aimed to investigate the role and potential mechanism of miR-27a during liver I/R injury. A liver I/R model was induced via 60 min of ischemia and reperfusion for 6 h in rats. Cells were transfected with miR-27a mimics or the miR-27a inhibitor to examine the effect of miR-27a on liver I/R. Apoptotic cells were detected by flow cytometry and TUNEL staining. The expression of miR-27a was measured by real-time PCR. The expression of peroxisome proliferator-activated receptor γ (PPARγ); gastrin-releasing peptide 78 (GRP78) and C/EBP homologous protein (CHOP) were detected by western blot analysis. The results showed that miR-27a was significantly upregulated during I/R injury in vivo and in vitro. In addition, miR-27a inhibitors attenuated hypoxia/reoxygenation (H/R)-induced oxidative stress, endoplasmic reticulum stress (ERS) and apoptosis in AML12 cells. By contrast, miR-27a mimics promoted hypoxia/reoxygenation-induced ERS, and apoptosis. Furthermore, PPARγ was identified as a target gene of miR-27a using bioinformatic analysis and a dual-luciferase reporter assay. Knockdown of PPARγ significantly abrogated the inhibitory effect of miR-27a inhibitors on the ERS pathway. Moreover, the miR-27a antagomir attenuated liver I/R injury in rats, a finding manifested by reduced ALT/AST, hepatocyte apoptosis, oxidative stress and inhibition of the ERS pathway. Taken together, these findings demonstrate that suppression of miR-27a protects against liver I/R injury by targeting PPARγ and by inhibiting the ERS pathway. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are a family of highly conserved, non-coding single-stranded RNAs transcribed as ~70 nucleotide precursors to an 18–22 nucleotide product (1). miRNAs can silence their homologous target genes at the post-transcriptional level, and these genes have been revealed to play an important role in tumorigenesis, invasion and metastasis (2). MicroRNA-27a (miR-27a), transcripted by miR-27a gene, has proved to implicate with many kinds of solid tumors, showing potential as a useful biomarker or drug target for clinical application. However, even though miR-27a has been reported in many cancers, the mechanism and signal pathways of miR-27 in oncogenesis, invasion, and metastasis are still obscure. Moreover, recent studies show that miR-27a pays an important role in epithelial-mesenchymal-transition, regulating tumor immune response, and chemoresistance. In this review, we summarize the current literature, demonstrate the established link between miR-27a and tumorigenesis, and focus on recently identified mechanisms. The review also aims to demonstrate the potential of miR-27a as a diagnostic and/or prognostic biomarker in solid tumors and to discuss the possibilities of targeted therapy and drug design. [ABSTRACT FROM AUTHOR]

Ischemia reperfusion injury (IRI) is one of the primary causes of acute renal injury and even acute renal failure. An increasing body of evidence has indicated that the aberrant expression of microRNAs (miRNA/miR) is closely associated with the progression of IRI. In the process of renal IRI, inflammatory reactions, cell adhesion and the death of renal tubular epithelial cells serve important roles. The present study investigated the expression of renal miRNAs following renal IRI in an attempt to identify the miRNAs that exert pivotal functions in renal IRI. The present study revealed that miR-27a, which was downregulated in IRI, and the 3′-untranslated region (UTR) of Toll-like receptor 4 (TLR4) have associated binding sites. The levels of TLR4, miR-27a and specific associated proteins in the renal tissues of gestational rats were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis, immunohistochemistry and western blotting. The associations between miR-27a and TLR4 were analyzed, and the regulatory effect of miR-27a on TLR4 was detected via a luciferase reporter gene assay, western blotting and RT-qPCR in vivo and in vitro. In addition, the present study demonstrated that miR-27a suppressed the expression of TLR4 by binding to the 3′UTR of TLR4. The overexpression of miR-27a downregulated the expression of TLR4, which in turn inhibited the inflammation, cell adhesion and cell death in IRI. Therefore, miR-27a inhibited inflammation in IRI by decreasing the expression of TLR4. [ABSTRACT FROM AUTHOR]

Across the globe, gastric cancer is a significant public health problem. This meta-analysis was conducted to investigate the association of microRNA-27a (miRNA-27a) rs895819 with gastric cancer risk. The search of databases updated on October 10, 2018 included Pubmed, Embase, Cochrane Library and Web of science. Odds ratio (ORs) and 95% confidence interval (CIs) were calculated to assess the risk of tumor. Overall meta-analysis suggested the miRNA-27a rs895819 was not related to the gastric carcinogenesis among all model including allele contrast (G vs A, pooled OR: 1.096, 95% CI: 0.962–1.249, P = 0.196), codominant model (GG vs AA, pooled OR: 1.124, 95% CI: 0.794–1.592, P = 0.590; AG vs AA, pooled OR: 1.101, 95% CI: 0.966–1.217, P = 0.060), dominant model (AG + GG vs AA, pooled OR: 1.123, 95% CI: 0.964–1.307, P = 0.136) and recessive model (GG vs AG + AA, pooled OR: 0.927, 95% CI: 0.673–1.278, P = 0.644). Interestingly, among different ethnicity group, significant relation between rs895819 and gastric cancer was observed in co-dominant model among Chinese population (AG vs AA, pooled OR: 1.158, 95% CI: 1.038–1.291, P = 0.008) but not some regions of European population (AG vs AA, pooled OR: 0.852, 95% CI: 0.632–1.148, P = 0.179). Our results find that rs895819 contributed to occurrence of gastric cancer in co-dominant model in Chinese population. • Rs895819 genotype (AG vs AA) might increase risk of stomach neoplasms among Chinese population. • Rs895819 genotype using sequencing was related to increased risk of stomach neoplasms under co-dominant and dominant models. • Updated articles were included in our work to draw effective conclusion on this issue which made our study more convincing. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To elucidate how microRNA-27a and FoxO1 regulate cardiac dysfunction in mice. MATERIALS AND METHODS: Expression levels of ANP, BNP, β-MHC, α-SMA, Fn1, and Periostin in myocardial tissues of 2-month-old and 8-month-old microRNA-27a-KO mice and agematched wild-type mice were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. Dual-luciferase reporter gene assay was conducted in H9C2 cells to verify the binding condition between microRNA-27a and FoxO1. By transfection of microRNA-27a mimics or inhibitor, FoxO1 expression in H9C2 cells was determined at the mRNA and protein levels. HW/BW [ratio of heart weight (mg) and body weight (mg)], HW/ TL [ratio of heart weight (mg) and tibial length (mm)], LVPWDT [left ventricular posterior wall diastolic thickness (mm)], LVEDD [left ventricular end-diastolic dimension (mm)], and FS (fractional shortening) in mice treated with or without FoxO1 inhibitor AS1842856 were accessed through echocardiography. RESULTS: MicroRNA-27a-KO mice had larger LVEDD, HW/BW, and HW/TL, but lower FS and LVPWDT than those of age-matched wildtype mice. Besides, higher levels of ANP, BNP, β-MHC, α-SMA, Fn1, and Periostin were observed in myocardial tissues of microRNA-27a-KO mice compared with those of age-matched wild-type mice. Dual-luciferase reporter gene assay revealed lower luciferase activity in H9C2 cells co-transfected with microRNA-27a mimics and wild-type FoxO1 than that of controls. The expression level of FoxO1 was negatively regulated by microRNA-27a in H9C2 cells at the mRNA and protein levels. After AS1842856 injection, HW/BW, HW/TL, and LVEDD in microRNA- 27a-KO mice markedly decreased, whereas FS and LVPWDT elevated. By comparison, AS1842856 injection did not influence cardiac development in wild-type mice. CONCLUSIONS: MicroRNA-27a knockout could induce cardiac dysfunction in mice through upregulating FoxO1 expression. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the significant role of β-elemene in mouse models of oxygen-induced retinopathy (OIR). C57BL/6J neonatal mice were used to establish OIR models. They were divided into four groups: Normoxia, OIR, OIR control and OIR-treated. Mice in the OIR group were exposed to 75±5% oxygen for 5 days and returned to a normal oxygen environment on postnatal day 12 (P12). The OIR treated group was intravitreally injected with 1 µl β-elemene on P12 and subsequently returned to a normal oxygen environment for 5 days (P12-P17). Retinas were obtained on P17. Retinal neovascularization (RNV) was detected using adenosine diphosphatase staining and analyzed by counting the nuclei of neovascular endothelial cells. Vascular endothelial growth factor (VEGF) expression was determined by reverse transcription-quantitative polymerase chain reaction, immunohistochemistry and western blot analysis. MicroRNA (miRNA/miR) microarrays were used to screen out differentially expressed miRNAs between the OIR and β-elemene-treated groups. Binding the 3′-untranslated region (UTR) of VEGF and miR-27a was confirmed using luciferase assays. It was found that high oxygen concentrations accelerated RNV and increased the number of preretinal neovascular cells; β-elemene treatment reduced these effects. VEGF mRNA and protein expression was higher in the OIR and OIR control groups, compared with the normoxia and OIR-treated groups. Further, it was shown that miR-22, miR-181a-1, miR-335-5p, miR-669n, miR-190b, miR-27a and miR-93 were upregulated in the OIR-treated group, and downregulated in the OIR group. The prediction websites TargetScan and miRanda revealed that VEGF contained a potential miR-27a binding site in its 3′-untranslated region (UTR). Luciferase assays demonstrated that miR-27a directly bound to the 3′-UTR of VEGF. In vitro experiments demonstrated that miR-27a inhibited VEGF expression. In addition, β-elemene treatment upregulate miR-27a expression in vivo and in vitro. When miR-27a expression was depleted by miR-27a inhibitor, the protective effect of β-elemene on RNV was eliminated. The present study demonstrated that β-elemene reduced RNV in mouse OIR models via miR-27a upregulation, leading to reduced VEGF expression. This finding may contribute to the development of novel therapeutic strategies for human retinopathy. [ABSTRACT FROM AUTHOR]

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.

Abstract Angiotensin II (Ang II) modulates VSMCs phenotypic switch that play a critical role in the cardiovascular diseases. MicroRNA-27a (miR-27a) has been proven to be involved in regulating vascular remodeling; however, the functional role of miR-27a in VSMCs in response to Ang II stimulation need to be elucidated. Cell proliferation and migration were measured by Cell counting kit-8 (CCK-8), BrdU incorporation and scratch wound assay in VSMCs transfected with miR-27a or its inhibitor. The target of miR-27a was confirmed using bioinformatics analysis and luciferase reporter assay. Ang II treatment time-dependently increased proliferation and migration of VSMCs accompanied with downregulation of α-smooth muscle-actin (α-SMA) and upregulation of miR-27a expression. Moreover, knockdown of miR-27a in VSMCs significantly attenuated Ang II-induced cell proliferation and migration, whereas this effect was aggravated by overexpression of miR-27a. A potential mechanistic analysis revealed that miR-27a directly targeted α-SMA, which mediated miR-27a-induced cell proliferation and migration. In conclusion, these results indicate that miR-27a acts as a novel regulator of Ang II-induced proliferation and migration by directly targeting α-SMA expression in VSMCs in vitro , and may be a potential therapeutic target for treating vascular diseases. Highlights • Ang II treatment increases miR-27a expression in VSMCs. • Modulating miR-27a influences proliferation and migration VSMCs. • miR-27a directly targets α-SMA. [ABSTRACT FROM AUTHOR]

Muscle fibre types can transform from slow-twitch (slow myosin heavy chain (MyHC)) to fast-twitch (fast MyHC) or vice versa. Leucine plays a vital effect in the development of skeletal muscle. However, the role of leucine in porcine myofibre type transformation and its mechanism are still unclear. In this study, effects of leucine and microRNA-27a (miR-27a) on the transformation of porcine myofibre type were investigated in vitro. We found that leucine increased slow MyHC protein level and decreased fast MyHC protein level, increased the levels of phospho-protein kinase B (Akt)/Akt and phospho-forkhead box 1 (FoxO1)/FoxO1 and decreased the FoxO1 protein level. However, blocking the Akt/FoxO1 signalling pathway by wortmannin attenuated the role of leucine in porcine myofibre type transformation. Over-expression of miR-27a decreased slow MyHC protein level and increased fast MyHC protein level, whereas inhibition of miR-27a had an opposite effect. We also found that expression of miR-27a was down-regulated following leucine treatment. Moreover, over-expression of miR-27a repressed transformation from fast MyHC to slow MyHC caused by leucine, suggesting that miR-27a is interdicted by leucine and then contributes to porcine muscle fibre type transformation. Our finding provided the first evidence that leucine promotes porcine myofibre type transformation from fast MyHC to slow MyHC via the Akt/FoxO1 signalling pathway and miR-27a. [ABSTRACT FROM AUTHOR]

BACKGROUND: Ovarian cancer (OC) is the fifth most common type of cancer in women worldwide. MiR-27a plays an important role in the development of ovarian cancer. However, the exact function and molecular mechanism of miR-27a in epithelial-mesenchymal transition (EMT) has not been thoroughly elucidated to date. METHODS: Quantitative real-time PCR (qRT-PCR) was used to determine the expression of miR-27a and FOXO1 mRNA in ovarian tissues and cells. The function of miR-27a in ovarian cancer was investigated through overexpression and knockdown of miR-27a in vitro. Wound healing and Transwell assays were performed to evaluate the migration and invasive capacity of the cells. A luciferase reporter assay was conducted to confirm the interaction between miR-27a and FOXO1. Western blotting was used to evaluate FOXO1, EMT and Wnt/ β -catenin relative protein expression. RESULTS: In our study, we found that the mRNA expression level of miR-27a was significantly higher in ovarian cancer tissues and in HO8910 and OV90 cells. Functional experiments showed that miR-27a overexpression potentiated the migration and invasion of HO8910 and OV90 cells, while miR-27a inhibition reduced the cells' migration and invasion. Moreover, miR-27a upregulated the expression of mesenchymal cell markers and downregulated the expression of epithelial cell markers, which were restored via silencing of miR-27a expression. Subsequently, miR-27a was found to directly target and suppress the expression of FOXO1. Finally, we demonstrated that miR-27a promoted the progression of ovarian cancer cells and induced the process of EMT via the Wnt/ β -catenin signalling pathway through inhibition of FOXO1. CONCLUSIONS: Taken together, these results indicate that targeting miR-27a and FOXO1 could represent a strategy for anticancer therapy in ovarian cancer. [ABSTRACT FROM AUTHOR]

Increasingly, evidence has revealed that aberrant microRNA (miRNA) expression is involved in breast cancer carcinogenesis and further progression, including metastasis. miRNA (miR)-27a was previously identified to be abnormally expressed and to serve pro-oncogenic functions in multiple human cancer types, including breast cancer. However, its functions and underlying mechanisms in breast cancer remain poorly understood. In the present study, it was demonstrated that miR-27a was significantly upregulated in breast cancer tissues and cell lines compared with their normal counterparts. Overexpression of miR-27a resulted in enhanced cell migration by inducing epithelial-to-mesenchymal transition, while its knockdown effectively reversed these cellular events. The present study additionally confirmed for the first time, to the best of our knowledge, that F-box and WD repeat domain containing 7 (FBXW7) is a downstream target gene of miR-27a in human breast cancer cells. FBXW7 is underexpressed in breast cancer tissues and cell lines, and is an independent positive factor for the overall survival rate of patients with breast cancer. Notably, the ectopic expression of FBXW7 may effectively suppress the epithelial-to-mesenchymal transition and migratory activity of breast cancer cells, in addition to reversing the cell migration mediated by miR-27a. Altogether, the results of the present study indicated the important function of miR-27a in regulating the metastasis of breast cancer in a FBXW7-dependent manner, and provide evidence for the potential application of miR-27a in breast cancer therapy. [ABSTRACT FROM AUTHOR]

Prostate cancer (PCa) exhibits a high incidence among men, but there is no effective and non‑invasive biomarker for the diagnosis of PCa, and the pathogenesis of PCa remains unclear. The present study identified that miR‑27a was significantly overexpressed in the tumor tissues and sera of patients with PCa. In addition, high serum levels of miR‑27a were correlated with poor survival in patients with PCa. Receiver‑operating characteristic curves analysis demonstrated that the serum levels of miR‑27a exhibited a high area under the curve value. Furthermore, miR‑27a mimics or inhibitors significantly promoted or repressed the proliferation of PCa cells, respectively. In addition, it was identified that the expression of Sprouty2 (SPRY2) was inversely correlated with the expression of miR‑27a in PCa tissues. The knockdown or overexpression of SPRY2 promoted or suppressed the proliferation of PCa cells, respectively, and the overexpression of SPRY2 inhibited the increased proliferation and cell cycle distribution of PCa cells mediated by miR‑27a mimics. Taken together, these data indicated that the serum levels of miR‑27a may be a novel and non‑invasive biomarker for the diagnosis and prognosis of patients with PCa, and miR‑27a/SPRY2 may be a therapeutic target for the treatment of PCa. [ABSTRACT FROM AUTHOR]

Acute lung injury (ALI) is a critical clinical condition with a high mortality rate, characterized with excessive uncontrolled inflammation and apoptosis. Recently, microRNAs (miRNAs) have been found to play crucial roles in the amelioration of various inflammation-induced diseases, including ALI. However, it remains unknown the biological function and regulatory mechanisms of miRNAs in the regulation of inflammation and apoptosis in ALI. The aim of this study is to identify and evaluate the potential role of miRNAs in ALI and reveal the underlying molecular mechanisms of their effects. Here, we analyzed microRNA expression profiles in lung tissues from LPS-challenged mice using miRNA microarray. Because microRNA-27a (miR-27a) was one of the miRNAs being most significantly downregulated, which has an important role in regulation of inflammation, we investigated its function. Overexpression of miR-27a by agomir-27a improved lung injury, as evidenced by the reduced histopathological changes, lung wet/dry (W/D) ratio, lung microvascular permeability and apoptosis in the lung tissues, as well as ameliorative survival of ALI mice. This was accompanied by the alleviating of inflammation, such as the reduced total BALF cell and neutrophil counts, decreased levels of tumor necrosis factor alpha (TNF-α), interleukin-1 (IL-6) interleukin-1β (IL-1β) and myeloperoxidase (MPO) activity in BAL fluid. Toll-like receptor 4 (TLR4), an important regulator of the nuclear factor kappa-B (NF-κB) signaling pathway, was identified as a novel target of miR-27a in RAW264.7 cells. Furthermore, our results showed that LPS stimulation increased the expression of MyD88 and NF-κB p65 (p-p65), but inhibited the expression of inhibitor of nuclear factor-κB-α (IκB-α), suggesting the activation of NF-κB signaling pathway. Further investigations revealed that agomir-miR-27a reversed the promoting effect of LPS on NF-κB signaling pathway. The results here suggested that miR-27a alleviates LPS-induced ALI in mice via reducing inflammation and apoptosis through blocking TLR4/MyD88/NF-κB activation. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate whether microRNA-27a (miRNA27a) is associated with the pathogenesis of diabetic retinopathy, and to elucidate the underlying molecular mechanism of any potential association. In retinal pigment epithelial (RPE) cells treated with high glucose, miRNA27a expression, determined by reverse transcription-quantitative polymerase chain reaction analysis, was decreased. Caspase-3/9 activity and B-cell lymphoma 2-associated X (Bax) protein expression was increased in RPE cells subjected to high glucose. Inhibition of miRNA27a suppressed the viability, and increased the caspase-3/9 activity and Bax protein expression of RPE cells treated with high glucose. Inhibition of miRNA27a expression also increased the expression of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α and Toll-like receptor 4 (TLR4) in RPE cells treated with high glucose. Treatment with immunostimulatory (is)RNA directed against TLR4 was observed to inhibit caspase-3/9 activity, decrease the expression of TLR4, Bax, IL-6, IL-1β and TNF-α, and increase the viability of RPE cells subjected to high glucose following the inhibition miRNA27a. In conclusion, the results of the present study suggest that miRNA27a protects RPE cells subjected to high glucose via inhibiting inflammation and apoptosis through targeting TLR4. [ABSTRACT FROM AUTHOR]

The present study aimed to determine the expression of vascular endothelial growth factor A (VEGFA) and microRNA (miRNA/miR)-27a in hippocampal tissues, and serum from a depression model of rats. In addition, the present study aimed to understand the mechanism of regulation of miR-27a in depression. A total of 40 male rats were selected, and divided into the control and depression model groups. The rats in the model group were subjected to 14 types of stimulations to model depression. By determining the body weight, syrup consumption rate and open field test score, the extent of depression in the rats was evaluated. Quantitative-polymerase chain reaction was used to determine the expression of VEGFA mRNA and miR-27a in hippocampal tissues, and serum. ELISA was used to measure the content of VEGFA protein in serum, while western blotting was employed to determine the expression of VEGFA protein in hippocampal tissues. A dual luciferase assay was carried out to identify the interactions between VEGFA mRNA and miR-27a. The rats in the depression model group showed depression symptoms and the depression model was successfully constructed. Rats with depression had lower VEGFA mRNA and protein expression in the hippocampus, and peripheral blood compared with the control group. Rats in the depression model group had reduced levels of miR-27a in the hippocampus and peripheral blood, which may be associated with the levels of VEGFA. miR-27a was able to bind with the 3′-untranslated region of VEGFA mRNA to regulate its expression. The present study demonstrated that miR-27a expression in hippocampal tissues and blood from rats with depression is upregulated, while the expression of VEGFA mRNA and protein is downregulated. miR-27a may participate in the pathological process of depression in rats by regulating VEGFA. [ABSTRACT FROM AUTHOR]

The present study investigated the role of microRNA (miR)‑27a in the development of arthritis and its mechanism of action. Initially, collagen was used to develop an in vivo rat model of arthritis. Changes in the miRs in the rats were analyzed. It was subsequently observed that miR‑27a expression was reduced in patients with arthritis, compared with the control group. In the present study an in vitro miR‑27a overexpression model of arthritis was established and it was observed that miR‑27a increased the proliferation of osteoblast‑like cells in vitro. miR‑27a overexpression promoted osteogenic differentiation, increased alkaline phosphatase (ALP) and osteoporosis (OST) content, induced insulin‑like growth factor binding protein-5 (IGFBP‑5) protein expression, reduced inflammation and suppressed peroxisome proliferator‑activated receptor γ (PPARγ) and matrix metalloproteinase-17 (MMP‑17) protein expression in arthritis. However, miR‑27a downregulation inhibited osteogenic differentiation, increased inflammation and PPARγ and MMP‑17 protein expression and suppressed ALP and OST content in an in vitro model of arthritis. The PPARγ inhibitor reduced the function of miR‑27a downregulation on arthritis. Therefore the results of the present study revealed that miR‑27a regulates arthritis via PPARγ. [ABSTRACT FROM AUTHOR]

The use of glucagon‑like peptide‑1 analogues, such as liraglutide, as hypoglycemic drugs has been widely employed in clinical practice. Liraglutide is reported to exert potential anti‑breast cancer effects, however the specific mechanisms of this action remain unknown. In the present study, MCF‑7 human breast cancer cells were cultured in vitro and treated with various concentrations of liraglutide. Cell Counting Kit‑8, colony formation and flow cytometry assays were performed to determine the proliferation and apoptosis of cells following treatment. Furthermore, reverse transcription‑quantitative polymerase chain reaction was employed to measure the expression level of microRNA (miRNA/miR)‑27a. In addition, miR‑27a mimics, inhibitors and negative controls were transfected into MCF‑7 cells and the proliferation and apoptosis of cells following transfection was subsequently determined. Western blotting was performed to detect alterations in the protein expression of AMP‑activated protein kinase catalytic subunit α2 (AMPKα2), proliferating cell nuclear antigen and cleaved‑caspase‑3 following treatments. The results demonstrated that, following treatment with liraglutide, the proliferation of MCF‑7 cells was reduced and the apoptosis was increased, compared with the control group; this effect was increased with increasing concentrations of liraglutide. In addition, liraglutide treatment downregulated miR‑27a expression in MCF‑7 cells. While the overexpression of miR‑27a promoted cell proliferation and inhibited apoptosis, knockdown of endogenous miR‑27a inhibited cell proliferation and promoted apoptosis in MCF‑7 cells. Furthermore, the expression of AMPKα2 protein in the group transfected with miR‑27a mimics was decreased, while it was increased in MCF‑7 cells transfected with miR‑27a inhibitors. In conclusion, liraglutide may have a role in the inhibition of proliferation and promotion of apoptosis in MCF‑7 cells. Concerning the mechanism of these effects, liraglutide may inhibit miR‑27a expression, which subsequently increases the expression of AMPKα2 protein. The present study provides an experimental basis for the clinical treatment strategies of T2DM patients with breast cancer. [ABSTRACT FROM AUTHOR]

Excessive inflammatory response and apoptosis play an important role in the sepsis-induced liver injury. Paclitaxel, a diterpene alkaloid of Taxus brevifolia , is widely used as an anti-tumor drug and shows protective effects on acute lung and kidney injury. However, whether it has a protective effect against sepsis-induced liver injury has not been reported. The objective of this study was to investigate the protective effects of paclitaxel in septic liver injury in mice and associated molecular mechanisms. Our results showed that paclitaxel treatment improved LPS-induced liver injury, as evidenced by the reduced aminotransferase activity, histological scores and apoptosis in the liver tissues. This was accompanied by the alleviating of inflammation and oxidative stress, such as decreased levels of tumor necrosis factor alpha (TNF-α), interleukin-1 (IL-6) interleukin-1β (IL-1β) and malondialdehyde (MDA) and increased levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) in serum and liver tissues. Subsequent microarray and qRT-PCR analysis further showed that miR-27a was significantly decreased in mice with sepsis, which was recovered by paclitaxel pretreatment. Antagomir-miR-27a suppressed the therapeutic effects of paclitaxel in mice liver injury model via promoting inflammatory response. Of note, TAB3, which participated in the activation of the NF-κB signaling pathway, was identified as a direct target of miR-27 by luciferase reporter gene assays. Then, we revealed a reverse relationship between miR-27a expression levels and TAB3 mRNA levels in liver tissues from septic mice. Furthermore, paclitaxel treatment significantly decreased the expression of NF-κB p65, but increased inhibitor of nuclear factor-κB-α (IκBα) protein levels in septic mice, suggesting the inactivation of NF-κB signaling pathway. Notably, the inhibitory effects of paclitaxel on NF-κB signaling pathway were reversed by antagomir-miR-27a. Our data indicated that paclitaxel significantly attenuated septic induced liver injury through reducing inflammatory response via miR-27a/TAB3/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Osteosarcoma is the most common type of malignant tumor arising from bone in children and adolescents. Accumulating evidences have shown the aberrant expression of numerous miRNAs is associated with the development and metastasis of osteosarcoma. The present study was conducted to investigate miR-27a expression in osteosarcoma tissues and cells. In the present study, quantitative RT-qPCR was used to measure the expression levels of miRNA and mRNA in osteosarcoma tissues and cells. Transwell assays were used to detect the effects of miR-27a on the invasive and migratory potential of cells. Luciferase reporter and western blot analysis were conducted to confirm cyclin G1 (CCNG1) as the target gene of miR-27a. The results showed that miR-27a was significantly upregulated in human osteosarcoma tissues and cell lines. The western blot analysis revealed that the overexpression of miR-27a suppressed CCNG1 protein expression. Luciferase reporter assays confirmed that CCNG1 is a direct target of miR-27a in osteosarcoma cells. The results suggest that miR-27a downregulates CCNG1 expression in osteosarcoma and acts as an oncogene directly targeting CCNG1. Thus, the miR-27a/CCNGI axis is a potential therapeutic target for human osteosarcoma. [ABSTRACT FROM AUTHOR]

The current study aimed to investigate the role of miR-27a in intervertebral disc degeneration (IDD) and to examine the underlying mechanisms. Quantitative polymerase chain reaction (qPCR) was performed to detect the expression level of miR-27a in the nucleus pulposus (NP) tissues of patients with IDD, and the results revealed an increasing expression of miR-27a in IDD compared with the control. To further investigate the role of miR-27a in IDD, a stable human NP cell line with low miR-27a expression was generated by transfecting cells with a lentiviral antigomiR-27a inhibitor. In addition, a human NP cell inflammation model was established by lipopolysaccharide (LPS; 10 µM) stimulation. The miR-27a expression in NP cells was determined by qPCR, while the expression of its target proteins; p-p38 and nuclear factor (NF-κB) was measured by western blot analysis. Furthermore, the mRNA and protein expression levels of proinflammatory factors, including interleukin (IL)-1ß, IL-6 and tumor necrosis factor-α (TNF-α), were also evaluated by qPCR and ELISA, respectively. The current results confirmed that miR-27a was significantly upregulated in IDD. In vitro, downregulation of miR-27a in LPS-stimulated NP cells by transfection with the miR-27a inhibitor resulted in suppression of p-p38 and NF-κB expression levels. Furthermore, the production of the proinflammatory factors IL-1ß, IL-6 and TNF-α was significantly reduced in LPS-stimulated NP cells with downregulated miR-27a. In conclusion, miR-27a may function as a promoter in IDD development, while inhibition of miR-27a may suppress proinflammatory factors released by intervertebral disc cells by regulating the p38/mitogen-activated protein kinase (MAPK) signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: This study aims to explore the effects of microRNA-27a (miR-27a) on the proliferation and invasiveness of colon cancer cells through the Secreted Frizzled-related protein 1 (SFRP1) and the Wnt/β-catenin signaling pathway. Methods: Colon cancer tissues and adjacent normal tissues from 125 colon cancer patients, together with the HCEpic, HCT-116, HT-29, SW480 and SW620 cell lines, were prepared for this study. The transfected HCT-116 cells were divided into the miR-27a mimics, miR-27a-NC, anti-miR-27a, blank, Lv-SFRP1, Lv-NC, and miR-27a mimics + Lv-SFRPl groups. RT-qPCR was performed to detect the expressions of miR-27a and SFRP1 mRNA. A dual-luciferase reporter assay was conducted to examine the effect of miR-27a on SFRP1. Western blotting was used to measure the expressions of the SFRP1, β-catenin, GSK-3β, p-β-catenin, p-GSK-3β, c-Myc and cyclin D1 proteins. MTT, soft agar clone formation and Transwell chamber assays were performed to detect cell proliferation and invasion. Results: Compared with normal tissues and cells, colon cancer tissues and cells demonstrated significantly higher expression of miR-27a, but lower expressions of SFRP1 mRNA and protein. MiR-27a negatively regulated the expression of SFRP1 mRNA. SFRP1 was also found to be a target gene of miR-27a. In the miR-27a mimic group, the proliferation and invasiveness of colon cancer cells were significantly increased, while the expressions of GSK-3β and p-β-catenin were remarkably down-regulated; in contrast, the expressions of p-GSK-3β, β-catenin, c-Myc and cyclin D1 were up-regulated. While the proliferation and invasiveness of colon cancer cells in the anti-miR-27a and Lv-SFRP1 groups were decreased, the expressions of GSK- 3β and p-β-catenin were elevated, and the expressions of p-GSK-3β, β-catenin, c-Myc and cyclin D1 were decreased. Conclusion: These findings indicated that miR-27a could promote the proliferation and invasiveness of colon cancer cells by targeting SFRP1 through the Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Huntington's disease (HD) is a fatal genetic disease caused by abnormal aggregation of mutant huntingtin protein (mHtt). Reduction of mHtt aggregation decreases cell death of the brain and is a promising therapeutic strategy of HD. MicroRNAs are short non-coding nucleotides which modulate various genes and dysregulated in many diseases including HD. MicroRNA miR-27a was reported to be reduced in the brain of R6/2 HD mouse model and modulate multidrug resistance protein-1 (MDR-1). Using subventricular zone-derived neuronal stem cells (NSCs), we used in vitro HD model to test the effect of miR-27a on MDR-1 and mHtt aggregation. R6/2-derived NSCs can be differentiated under condition of growth factor deprivation, and the progression of differentiation leads to a decrease of MDR-1 level and efflux function of cells. Immunocytochemistry result also confirmed that mHtt aggregation was increased with differentiation. We transfected miR-27a in the R6/2-derived differentiated NSCs, and examined phenotype of HD, mHtt aggregation. As a result, miR-27a transfection resulted in reduction of mHtt aggregation in HD cells. In addition, MDR-1, which can transport mHtt, protein level was increased by miR-27a transfection. Conversely, knock-down of MDR-1 through MDR-1 siRNA increased mHtt aggregation in vitro . Our results indicate that miR-27a could reduce mHtt level of the HD cell by augmenting MDR-1 function. [ABSTRACT FROM AUTHOR]

Objective To investigate the role of MicroRNA-27a (miR-27a) in the anti-apoptotic effects of liraglutide (LR) in cardiomyocytes. Methods The H9c2 rat cardiomyocytes were cultured in different environment and divided into low glucose group, high glucose group, miR-27a over-expression group, miR-27a inhibited group, miR-27a negative over-expression group, miR-27a negative inhibited group, liposome 2000 group, hypertonic solution group, LR intervention group, LR intervention and miR-27a over-expression group, LR intervention and miR-27a negative over-expression group. The expression of miR-27a in cardiomyocytes, which were cultured in low glucose solution, high glucose solution and interferenced by LR, was detected by fluorescence-based real-time quantitative polymerase chain reaction (PCR). The apoptosis of cardiomyocytes was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and then the cell apoptosis index (AI) was computed and compared among the groups. The expression of apoptosis-related protein B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2 associated X protein (Bax), cysteine-containing aspartate-specific proteases-3 (caspase-3), cellular inhibitors of apoptosis-1 (cIAP-1), anti-apoptotic protease activating factor-1 (Apaf-1) was detected by Western blotting method. Data were analyzed with one-way analysis of variance test or t test. Results The expression of miR-27a in high glucose group was higher than that in low glucose group(35.8±4.9 vs 1.0±0.0, t=12.199, P< 0.05). Compared with the high glucose group, the AI of cardiomyocytes increased significantly in miR-27a over-expression group (78.5±6.6 vs 22.9±1.7, least-significant difference (LSD), P<0.05), and the protein expression of Bcl-2 and cIAP-1 decreased (LSD, both P<0.05), the protein expression of Bax, caspase-3 and Apaf-1 increased(LSD, all P<0.05). Compared with the high glucose group, the AI of cardiomyocytes decreased significantly in miR-27a inhibition group (2.6±0.4 vs 22.9±1.7, LSD, P<0.05), and the protein expression level of Bcl-2 and cIAP-1 increased (LSD, both P<0.05), the protein expression level of Bax, caspase-3 and Apaf-1 significantly decreased(LSD, all P<0.05). Compared with the high glucose group, the expression level of miR-27a decreased significantly in LR intervention group (7.9 ± 0.2 vs 35.8 ± 4.9, t=-9.785, P<0.05), and the AI of cardiomyocytes decreased remarkably (4.9±0.1 vs 22.9±1.7, LSD, P<0.05). The AI of cardiomyocytes increased in LR intervention and miR-27a over-expression group when compared with that in LR intervention group (61.3±5.1 vs 4.9±0.1, LSD, P<0.05). Conclusion LR may ameliorate the apoptosis of cardiomyocytes in the condition of high glucose by inhibiting the expression of miR-27a in H9c2 rat cardiomyocytes. [ABSTRACT FROM AUTHOR]