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Objective To verify that miR-203 plays a significant role in promoting M2 macrophage polarization through targeting 3'-UTR of TLR4 mRNA and therefore inhibiting TLR4-Myd88-NF- κB signaling. We speculated the specific binding of miR-203 and 3'-UTR of TLR4 mRNA by miRanda software. And we constructed plasmid pmirGLO-TLR4 3'-UTR and its mutant plasmid pmirGLO-mut-TLR4 3'-UTR based on the sequence of 3'-UTR of TLR4 mRNA. Cell line 293T was co-transfected with plasmids pmirGLO-TLR4 3'-UTR or pmirGLO-mut-TLR4 3'-UTR and mmu-miR-203-3p mimics or mimic NC. Dual-luciferase reporter assay systemwas used to detect the specific binding of miR-203 and 3'-UTR of TLR4 mRNA. The expressions of M1 macrophage markers (iNOS, TNF-琢, CCL-3, IL-23), M2 macrophage markers (Arg-1, CX3CR1, IL-4, MRC, IL-10 and Ym1) were detected by Real-time PCR. And the expression of TLR4-Myd88-NF-κB signaling was detected by Real-time PCR and Western blot. The antagonist of TLR4, TAK-242, was used to further confirmed the role of miR-203 in regulating M2 macrophage polarization and inhibiting the TLR4-Myd88-NF-κB signaling pathway. The Relative luciferase activity was significantly lower in 293T cells co-transfected with pmirGLO-TLR4 3'-UTR plasmid and mmu-miR-203-3p mimics than in 293T cells co-transfected with either pmirGLO-mut-TLR4 3'-UTR plasmid or mimic NC. Real-time PCR and Western blot revealed that the expressions of M1 macrophage markers (iNOS, TNF-琢, CCL-3, IL-23) were significantly decreased and the expressions of M2 macrophage markers (Arg-1, CX3CR1, IL-4, MRC, IL-10 and Ym1) were signifcantly increased in mouse macrophage RAW264.7 transfected with mmu-miR-203-3p mimics when compared with control group or mouse macrophage RAW264.7 transfected with mimic NC, accompanied by downregulation of TLR4-Myd88-NF-κB signaling. And this phenomenon was abrogated by the inhibiting of TLR4-Myd88-NF-κB signaling pathway in mouse macrophages RAW264.7 treated with TAK-242. Conclusion Taken together, miR-203 could promote M2 macrophages polarization through directly targeting 3'-UTR of TLR4 mRNA and therefore inhibiting the TLR4-Myd88-NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The purpose of this study was to investigate how miR-203 promotes osteogenic differentiation of bone marrow mesenchymal cells (BMSCs) by regulating its target gene DKK1, thereby inhibiting the occurrence of osteoporosis. PATIENTS AND METHODS: A total of 60 cases with postmenopausal osteoporosis and 40 cases of normal individuals were recruited. The expression of miR-203 in serum of all cases was detected by quantitative reverse transcriptase- polymerase chain reaction (qRT-PCR). The capacity of osteogenesis and adipogenic differentiation of MSCs was determined by alizarin red staining and oil red staining, respectively. Transfection of miR-203 mimics and miR- 203 inhibitor were mediated by Liposomes, and then the MSCs were induced osteogenic and adipogenic differentiation. MiR-203 mimic was co-transfected with wild-type or mutant DKK1 for luciferase reporter gene detection. In the osteoporosis model of rats, the tibia was taken for micro-CT examination of bone mineral density (BMD) and bone volume/structural parameters (BV/TV), while the femur was taken for the measurement of absorption parameters (Ob.S)./BS) and the number of osteoclasts per circumference of bone (N.Oc/B.Pm). RESULTS: The expression level of miR-203 was significantly lower in patients with postmenopausal osteoporosis than that in normal individuals. The osteogenic capacity of BMSCs in these patients was reduced, while their adipogenic capacity was enhanced. MiR-203 promoted the expression of osteogenic genes and inhibited that of adipogenic genes. Knockdown of miR-203 decreased the level of osteogenic related related genes but increased that of adipogenic related genes, while overexpression of miR-203 led to the opposite results. Furthermore, miR- 203 inhibited the protein expression of DKK1. In addition, bone density and bone volume/structural parameters were lower in ovariectomized rats than those in normal rats. Meanwhile, bone resorption parameters and the number of osteoclasts per bone circumference in ovariectomized rats were higher than those in normal rats. CONCLUSIONS: MiR-203 can promote osteogenic differentiation of mesenchymal stem cells by downregulating the gene expression of DKK1. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate the relationship between microRNA-203 (miR-203) and diabetic nephropathy and its potential mechanism. MATERIALS AND METHODS: The expression of microRNA-203 in mice with diabetic nephropathy and M4200 cells cultured with high glucose was detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Toll-like receptor 4 (TLR4), the target gene of microRNA-203, was predicted and screened by bioinformatics method. Real-time quantitative PCR and Western blot were used to detect the endogenous TLR4 level in renal cortex of db/db mice with diabetic nephropathy and glomerular mesangial cells cultured in high glucose or low glucose. The expression of microRNA-203 and TLR4 mRNA were evaluated by RT-PCR after treatment of miR-203 mimics and inhibitor. The protein of TLR4 level was detected by Western blot. Additionally, the proliferation ability of cells was evaluated by Cell Counting Kit-8 (CCK8). The target relationship between microRNA-203 and TLR4 3' UTR was confirmed by luciferase reporter assay RESULTS: The expression of miR-203 was significantly decreased in the kidney of mice with diabetic nephropathy and M4200 cells cultured in high glucose. On the contrary, TLR4 expression was significantly increased. Results of in vitro experiments showed that miR-203 could bind to 3'UTR region of TLR4. Overexpression of microRNA-203 significantly decreased the levels of TLR4 mRNA and protein. Meanwhile, low expression of miR-203 leaded to increased TLR4 expression, resulting in an enhanced proliferation of M4200 cells. CONCLUSIONS: The downregulation of microRNA-203 leaded to an increased level of TLR4, thus promoting proliferation of M4200 cells in the pathogenesis of diabetic nephropathy. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate whether miR-203 is involved in the osteogenic differentiation of rat mesenchymal stem cells (MSCs) by regulating DDK1, thus participating in the pathogenesis of osteoporosis. PATIENTS AND METHODS: miR-203 expression in serum samples of 60 osteoporosis patients and 60 normal subjects was detected using Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) assay. MSCs were isolated from bone marrow of rats and then identified. Subsequently, the effects of miR-203 and DKK1 on osteogenic differentiation were estimated by alkaline phosphatase (ALP) activity, alizarin red staining, ALP staining, respectively. Expression levels of osteogenic-specific genes were detected by Western blot. Rescue experiments were conducted to confirm whether miR-203 could promote osteogenic differentiation of MSCs by inhibiting DKK1. RESULTS: Serum level of miR-203 in osteoporosis patients was significantly lower than that of the normal subjects. Overexpressed miR-203 in MSCs enhanced ALP activity, expression of osteogenic marker genes and the number of calcified cells. Additionally, miR-203 could bind to DKK1. The regulatory effect of miR-203 on osteogenic differentiation in MSCs was reversed by DKK1. CONCLUSIONS: MiR-203 promotes the differentiation of rat MSCs into osteoblast-like cells, which may be associated with the regulation of DKK1 expression. [ABSTRACT FROM AUTHOR]

Rapid proliferation is one of the critical characteristics of breast cancer. However, the underlying regulatory mechanism of breast cancer cell proliferation is largely unclear. The present study indicated that sevoflurane, one of inhalational anesthetics, could significantly suppress breast cancer cell proliferation by arresting cell cycle at G1 phase. Notably, the rescue experiment indicated that miR-203 was upregulated by sevoflurane and mediated the function of sevoflurane on suppressing the breast cancer cell proliferation. The present study indicated the function of the sevoflurane/miR-203 signaling pathway on regulating breast cancer cell proliferation. These results provide mechanistic insight into how the sevoflurane/miR-203 signaling pathway supresses proliferation of breast cancer cells, suggesting the sevoflurane/miR-203 pathway may be a potential target in the treatment of breast cancer. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Extra-cellular signal regulated kinase (ERK)/mitogen activated protein kinase (MAPK) signaling pathway is widely involved in cell proliferation, apoptosis, and drug resistance. MAPK kinase 1 (MEK1) is the upstream protein kinase of ERK that can activate ERK/MAPK signaling pathway. microRNA 203 (MiR-203) down-regulation is found to be associated with prostate cancer pathogenesis. Bioinformatics analysis showed the complementary targeted relationship between miR-203 and the 3'-UTR of MEK1 mRNA. This study explored the role of miR-203 in regulating prostate cancer cell proliferation, apoptosis, and ADM resistance through affecting MEK1 expression. MATERIALS AND METHODS: Dual luciferase assay confirmed the targeted relationship between miR-203 and MEK1. MiR-203, MEK1, p-ERK1/2, and B cell lymphoma 2 (Bcl-2) expressions were compared in normal prostate epithelial cells PrEC, prostate cancer cells PC-3M, and drug resistance cells PC-3M/ADM. PC-3M, PC-3M/ADM cell apoptosis and proliferation were detected by using flow cytometry under ADM treatment at IC50 concentration of PC-3M cells. PC-3M cells were cultured in vitro and divided into four groups, including microRNA-normal control (miR-NC), miR-203 mimic, small interfere NC (si-NC), and si-MEK1. RESULTS: MiR-203 targeted and inhibited MEK1 expression. MiR-203 levels and cell apoptosis were significantly lower, while MEK1, p-ERK1/2, Bcl-2, and cell proliferation were significantly higher in PC-3M/ADM cells compared to the PC-3M cells. MiR-203 mimic and/or si-MEK1 transfection significantly reduced MEK1, p-ERK1/2, and Bcl-2 levels, attenuated cell proliferation, induced cell apoptosis, and decreased drug resistance. CONCLUSIONS: MiR-203 elevation suppressed prostate cancer PC-3M cell proliferation, promoted apoptosis, and weakened ADM resistance through targeted inhibiting MEK1 expression to alleviate ERK/MAPK signaling pathway and Bcl-2 expression. [ABSTRACT FROM AUTHOR]

Increasing evidence has indicated that long noncoding RNAs (lncRNAs) play crucial roles in various biological processes, including glioma. However, the underlying mechanism of lncRNAs in gliomagenesis is still ambiguous. In this study, we aim to investigate the role of long intergenic noncoding RNA 00958 (LINC00958) in the tumorigenesis of glioma. Results revealed that LINC00958 was significantly upregulated in glioma tissues and cell lines compared with that of adjacent normal brain tissues and normal human astrocytes. Moreover, the ectopic overexpression of LINC00958 was correlated with poor prognosis of glioma patients. Loss-of-function experiments indicated that LINC00958 knockdown suppressed glioma cell proliferation, invasion, and induced cycle arrest at G0/G1 phase in vitro, and inhibited tumor growth in vivo. Bioinformatics programs and luciferase reporter assay revealed that miR-203 shared complementary binding sites with both 3′-untranslated region of LINC00958 and CDK2. In summary, our study concludes that LINC00958 acts as an oncogenic gene in the gliomagenesis through miR-203-CDK2 regulation, providing a novel insight into glioma tumorigenesis. [ABSTRACT FROM AUTHOR]

Introduction: MicroRNAs (miRNAs) are short non-coding single-stranded RNAs. Involving in post-transcriptional gene silencing, miRNAs are thought to play important roles in many cancers such as breast cancer. Paclitaxel is used widely in the treatment of breast cancer. In this study, we investigated the effect of paclitaxel treatment on the expression levels of two oncomirs (oncomiRs), miR-21 and miR-203, in breast cancer cell lines. Materials and methods: MTT assay was performed to determine IC50 of paclitaxel for human breast cancer cell lines including MCF-7, MDA-MB-231, SKBR3 and BT-474. After RNA extraction and cDNA synthesis, the expression levels of miRNAs were then quantitatively evaluated using real-time PCR. Results: Our results showed that after treatment, the expression levels of both miR-21 and miR-203 were significantly increased in HER2-positive cell lines, BT-474 and SKBR3. HER2-negative cell lines, MCF-7 and MDA-MB-231, in contrast had significantly decreased expression of both assessed oncomiRs. Conclusion: Our results showed that the expression levels of oncomiRs were increased in HER-2 positive breast cancer cells and this finding is in line with previous studies. Our findings present a probable mechanism of resistance against paclitaxel chemotherapy in HER2-positive breast cancers. [ABSTRACT FROM AUTHOR]

Alcoholic liver disease (ALD) is a global liver disease which characterized by liver inflammation, fatty liver, alcoholic hepatitis, or liver cirrhosis. Alcohol abuse is one of the main reasons for liver disease. Alcoholic fatty liver (AFL) disease is the early stage of ALD and associated with the excessive lipids accumulation in hepatocytes as well as oxidative stress. MicroRNA-203 (miR-203) is known to suppress the proliferation and metastasis of hepatocellular carcinoma, but the role in the progression of alcoholic liver disease is not clear and is warranted for further investigation. In the present study, we have found the expression of miR-203 is down-regulated in Gao-Binge alcoholic mice model and ethanol-induced AML-12 cell lines in vitro. Furthermore, over-expression of miR-203 decrease the lipids accumulation in liver and ethanol-induced AML-12 cells. Mechanistically, we identified that Lipin1 is a key regulator of hepatic lipid metabolism, and acts as a downstream target for miR-203. In summary, our results suggested that over-expression of miR-203 inhibited the liver lipids accumulation and the progression of AFL by targeting Lipin1. [ABSTRACT FROM AUTHOR]

Angiogenesis during placentation is of great significance in maintaining normal pregnancy. However, the molecular mechanisms of this process are not clear. It has been reported that miR-203 plays a critical role in the development and progression of many tumors but not focused on the relationship between miR-203 and placental angiogenesis. The present study aims to illustrate the correlation between miR-203 and vascular endothelial growth factor (VEGFA)/vascular endothelial growth factor receptors 2 (VEGFR2) in human placenta and human umbilical vein endothelial cells (HUVECs) obtained from 40 samples. Samples of human placenta were collected based on gestation age, which was divided into early preterm (n = 10), late preterm (n = 12), and term (n = 18). In this work, we demonstrated that the expression of miR-203 decreased significantly in the placenta according to the gestation age, in contrast, the expression of VEGFA and VEGFR2 increased accordingly. In vitro experiments revealed that overexpression of miR-203 not only suppressed the proliferation, migration, invasion, and tube formation of HUVECs but also affected the expression of VEGFA and VEGFR2. Furthermore, inhibition of miR-203 expression showed equally apparent positive effects on HUVECs. In conclusion, our study suggests that miR-203 plays an important role in regulating placental angiogenesis through inhibiting the expression of VEGFA and VEGFR2, thus miR-203 may represent a potential therapeutic target for patients with abnormal formation of blood vessels in the placenta. [ABSTRACT FROM AUTHOR]

Background The clinical significance of miR-203 and its prognostic value have not been investigated in gastric cancer. Methods We assessed miR-203 expression in 141 gastric cancer samples and 141 paired non-cancerous samples by real-time PCR and calculated using the 2 −ΔΔCt method. Differences between groups were examined for statistical significance by Student’s t -test. Survival curves were computed by the Kaplan-Meier method, and differences between survival curves were compared by the log rank test. Results The expression of miR-203 was significantly lower in gastric cancer samples compared to non-cancerous samples (P < 0.0001). Low miR-203 expression was found to be closely correlated with advanced stage (p = 0.005), and lymph node involvement (p = 0.009). Kaplan-Meier analysis with the log-rank test indicated that low miR-203 expression had a significant impact on overall survival (39.4% vs. 62.5%; P = 0.043) and progression-free survival (32.5% vs. 58.6%; P = 0.023). Furthermore, multivariate analysis revealed that miR-203 expression level was independent prognostic factors for overall survival (HR = 2.73, 95% CI: 1.69–8.91; P = 0.01), as well as progression-free survival (HR = 4.19, 95% CI: 2.91–10.12; P = 0.005). Conclusion Our data validate an important clinical significance of miR-203 in gastric cancer, and reveal that it might be a potential prognostic factor for gastric cancer. Large- scale and long-term follow-up studies are needed to confirm the significance of miR-203 in gastric cancer. [ABSTRACT FROM AUTHOR]

Involvement of the RGS17 oncogene in the promotion of non-small-cell lung cancer ( NSCLC) has been reported, but the regulation mechanism in NSCLC remains unclear. Micro RNAs (mi RNAs) negatively regulate gene expression, and their dysregulation has been implicated in tumorigenesis. To understand the role of mi RNAs in Regulator of G Protein Signaling 17 ( RGS17)-induced NSCLC, we showed that miR-203 was downregulated during tumorigenesis, and inhibited the proliferation and invasion of lung cancer cells. We then determined whether miR-203 regulated NSCLC by targeting RGS17. To characterize the regulatory effect of miR-203 on RGS17, we used lung cancer cell lines, A549 and Calu-1, and the constructed miR-203 and RGS17 overexpression vectors. The CCK8 kit was used to determine cell proliferation, and the Transwell® assay was used to measure cell invasion and migration. RT- PCR, western blots, and immunofluorescence were used to analyze expression of miR-203 and RGS17, and the luciferase reporter assay was used to examine the interaction between miR-203 and RGS17. Nude mice were used to characterize in vivo tumor growth regulation. Expression of miR-203 inhibited proliferation, invasion, and migration of lung cancer cell lines A549 and Calu-1 by targeting RGS17. The regulatory effect of miR-203 was inhibited after overexpression of RGS17. The luciferase reporter assay showed that miR-203 downregulated RGS17 by direct integration into the 3′- UTR of RGS17 mRNA. In vivo studies showed that expression of miR-203 significantly inhibited growth of tumors. Taken together, the results suggested that expression of miR-203 inhibited tumor growth and metastasis by targeting RGS17. [ABSTRACT FROM AUTHOR]

OBJECTIVE: MiR-181a plays a critical role in modulating T cell and B cell differentiation, as well as immune response. Its abnormal expression probably participates in the pathogenesis of systemic lupus erythematosus (SLE). MiR-203 is involved in regulating Toll-like receptor and inducing immune tolerance. Abnormal expression or function of miR-203 is related to multiple auto-immune diseases but its role in SLE remains unclear. This study, thus, investigated the serum level of miR-181a and miR-203, to analyze their roles in diagnosing and evaluating SLE. PATIENTS AND METHODS: SLE patients were recruited from our hospital, and divided into non-active and active SLE based on disease activity index, along with healthy individuals. qRT-PCR was used to quantify the serum miR-181a and miR-203 expression, and their correlation with clinical features. ROC was used to evaluate the diagnostic value on SLE, while survival curves were compared to show progression- free survival (PFS) between populations with high and low expression. RESULTS: SLE patients had significantly higher serum levels of miR-181a and lower miR-203, both of which were correlated with SLE activity. Expression levels of miR-181a and miR-203 were correlated with erythrocyte sedimentation rate, C reactive protein, anti-dsDNA antibody, complements, and SLEDAI score. Their expression levels had certain values in the differential diagnosis for active SLE (AUC=0.885 and 0.843). PFS in miR-181a high-expression individuals was lower than that in the low-miR-181 group (Χ²=7.474, p=0.029). Whilst, miR-203 high-expression SLE patients had higher PFS than low-expression group (Χ²=4.367, p=0.037). CONCLUSIONS: SLE patients had higher miR-181a and lower miR-203 expression, which thus may have critical implications in disease diagnosis and evaluation. [ABSTRACT FROM AUTHOR]

Background: Endometrial cancer (EC) is currently considered the fourth most frequent female cancer in Europe. In an attempt to achieve an early diagnosis, many studies have identified some putative biomarkers for gynecologic cancers, including circulating microRNAs (miRs) and aberrant promoter methylation status. Previous studies which have investigated miR-203 expression profiles in EC tissues and normal endometrial tissues concluded that miR-203 is regulated by methylation promoter. The aim of this study was to investigate the expression of miR-203 and promoter methylation levels in serum of EC patients and healthy controls (HC). Methods: Forty-five EC patients (64 ± 12 years) and 30 HC (63 ± 13 years) were enrolled before undergoing therapeutic procedures. RNA extraction from serum was performed with mirVana PARIS Isolation Kit (Thermo Scientific). miR expression was assessed by quantitative RT-PCR (Applied Biosystems). The expression levels of miR were normalized to miR-16 and calculated using the 2-ΔCt method. A quantitative methylation-specific PCR (MSP) technique was used to analyze miR-203 promoter methylation status. Differences between groups were assessed by Mann-Whitney test (for continuous variables) and chi-squared test (for categorical variables), whereas the correlation was calculated using Spearman's test. The diagnostic performance of miR-203 was defined using receiver operator characteristic (ROC) curves. Results: Serum expression levels of miR-203 were higher in EC patients compared to HC (p = 0.002). Aberrant miR-203 methylation was detected in 11/45 (24.4%) EC patients and in 2/30 (6.6%) HC (p = 0.046). The expression levels of miR-203 were not significantly correlated with promoter methylation status. The area under the curve of miR-203 expression was 0.71 (p = 0.002). Conclusions: The high circulating miR-203 expression levels in EC patients compared to HC confirm the role of this miR as a potential biomarker for diagnosis of EC. Aberrant miR-203 methylation assessed in the peripheral blood does not apparently reflect cancer biology. [ABSTRACT FROM AUTHOR]

Dmrt2b (doublesex and mab-3 related transcription factor 2b) has been revealed to be involved in zebrafish slow muscle development. However, the function of dmrt2a , a paralogue gene of dmrt2b , remains unclear during zebrafish muscle development. Here, we demonstrated that knockdown of dmrt2a resulted in severe developmental defects, and caused downregulation of fast muscle marker myhz-2 and upregulation of slow muscle marker myhz-5 , respectively. It is known that microRNAs (miRNAs) control many biological events including muscle development. Dmrt2a was predicted to be a target gene of miR-203, which was further verified by luciferase reporter assay, since miR-203a was found to directly reduce the expression of dmrt2a by binding to the seed sequence of its 3′UTR. After miR-203a injection into zebrafish embryos, the expression of dmrt2a was significantly inhibited. Similar to the effect of dmrt2a knockdown, miR-203a overexpression led to downregulation of myhz-2 and upregulation of myhz-5 . Our studies indicated that miR-203a directly regulated dmrt2a expression to control fast and slow muscle differentiation, while overexpression of miR-203a or knockdown of dmrt2a will impair fast muscle development and promote slow muscle development. [ABSTRACT FROM AUTHOR]

BACKGROUND: MicroRNAs (miRNAs) have been demonstrated to play an important role in the development and progression of various types of cancer including glioblastoma (GBM). OBJECTIVE: The aim of this study was to investigate the expression pattern and prognostic significance of serum miR-203 in patients with GBM. METHODS: miR-203 extracted from cell culture medium and serum samples was detected by real-time PCR. The correlation between serum miR-203 expression as well as clinicopathological characteristics and patient survival was determined. RESULTS: The expression level of miR-203 was remarkably reduced in the GBM cells and their culture medium. Serum miR-203 expression was significantly decreased in GBM patients compared with low grade glioma (LGG) patients and healthy controls. In addition, serum miR-203 discriminated GBM patients from LGG patients and healthy subjects. Chi-squared analysis showed that a significant correlation was found between low serum miR-203 expression and larger tumor size as well as lower Karnofsky Performance Scale scores. Patients with lower serum miR-203 suffered poorer overall survival (OS) and progression free survival (PFS). Multivariate analysis indicated that low miR-203 expression is an independent prognostic factor for poor OS in GBM patients. CONCLUSIONS: These data demonstrate that serum miR-203 expression might serve as a potential prognostic indicator of GBM. [ABSTRACT FROM AUTHOR]

Activation of hepatic stellate cells (HSCs) is a pivotal event during hepatic fibrogenesis. Activated HSCs are the main source of collagen and other extracellular matrix (ECM) components, and emerging antifibrotic therapies are aimed at preventing ECM synthesis and deposition. MicroRNAs (miRNAs) have been demonstrated to exert regulatory effects on HSC activation and ECM synthesis. In the present study, the HSC-T6 rat hepatic stellate cell line was transiently transfected with a miRNA (miR)-203 mimic, which is an artificial miRNA that enhances the function of miR-203, with a miR-203 inhibitor or with a scramble miRNA negative control. mRNA and protein expression levels of collagen (COL) 1A1, COL3A1, a-smooth muscle actin (a-SMA) and mothers against decapentaplegic homolog 3 (SMAD3) were assessed using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. The interaction between miR-203 and the 3'-untranslated region (UTR) of SMAD3 mRNA was examined using a dual-luciferase reporter assay. The proliferative capabilities of activated HSCs were measured using an MTT assay. The present results demonstrated that the mRNA and protein expression levels of COL1A1, COL3A1, a-SMA and SMAD3 were significantly upregulated following transfection of HSC-T6 cells with the miR-203 inhibitor. Conversely, COL1A1, COL3A1, a-SMA, and SMAD3 mRNA and protein expression appeared to be downregulated in rat HSCs transfected with miR-203 mimics. Notably, the inhibition of miR-203 expression was revealed to promote HSC proliferation, whereas increased miR-203 expression suppressed the proliferative capabilities of HSC-T6 cells. Furthermore, SMAD3 was revealed to be a direct target of miR-203. The present study suggested that miR-203 may function to prevent the synthesis and deposition of ECM components, including COL1A1, COL3A1 and a-SMA, and to inhibit the proliferation of HSCs through a SMAD3-dependent mechanism. Therefore, it may be hypothesized that miR-203 has potential as a novel target for the development of alternative therapeutic strategies for the treatment of patients with hepatic fibrosis in clinical practice. [ABSTRACT FROM AUTHOR]

Dysregulation of miR-203 by promoter methylation is associated with the development of various cancers. We aimed to explore the underlying link between promoter methylation and miR-203 expression in Kazakh esophageal squamous cell carcinoma (ESCC). MassARRAY® System spectrometry was used to quantitatively analyze the DNA methylation of 32 CpG sites within miR-203 in 99 Kazakh ESCC and 46 normal esophageal tissues (NETs) with similar population characteristics. We conducted real-time PCR to detect miR-203 expression levels and evaluated their association with methylation. Eleven CpG units within miR-203 promoter were frequently hypermethylated in ESCC compared with NETs(P < 0.05). The hypermethylation of several CpG units positively correlated with age, lower esophagus, constrictive type of ESCC, and moderately differentiated ESCC. Given the involvement of human papillomavirus (HPV) in etiology of ESCC was confirmed from our previous reports, herein we found that CpG units within miR-203 in HPV16-positive ESCC are more heavily methylated. Furthermore, miR-203 expression showed a nearly 4.5-fold decrease in ESCC than NETs (0.206 ± 0.336 vs. 0.908 ± 1.424,P< 0.001) and was significantly associated with lymph node metastasis(P= 0.012). The expression of miR-203 with 11 completely hypermethylated CpG units was approximately 6.5-fold lower than that with at least 1 unmethylated CpG unit(P< 0.001) and especially the CpG_15.16 and CpG_31.32 with higher methylation levels in ESCC tissues exhibited lower expression levels of miR-203, which indicated a reverse association between miR-203 methylation and expression. Hypermethylated miR-203 is a potential biomarker and targeted delivery of miR-203 could therefore serve as a preventive or therapeutic strategy for Kazakh ESCC. [ABSTRACT FROM PUBLISHER]

We sought to systematically evaluate the diagnostic and prognostic value of miR-203 in patients with colorectal cancer. To explore the diagnostic performance of miR- 203, eligible studies were identified from biomedical databases. Based on these results, 11 studies were pooled and included in this meta-analysis. The pooled sensitivity, specificity, and diagnostic odds ratios of miR-203 were 0.83 (95% confidence interval, CI: 0.78-0.86), 0.80 (95% CI: 0.77-0.83), and 19.27 (95% CI: 7.23-51.36) for the diagnosis of colorectal cancer. The area under the curve for miR-203 for diagnosing colorectal cancer was 0.89. Patients with higher expression of tissue miR-203 had poor overall survival (pooled hazard ratio: 1.63; 95% CI: 1.03-2.57, P=0.04), but serum miR-203 was not predictive (pooled hazard ratio: 1.59; 95% CI: 0.31-8.12, P=0.58). The miR-203 values of tissue and serum merged together may perhaps predict superior overall survival (pooled hazard ratio: 1.62; 95% CI: 0.93-2.82), but the effect was not significant (P=0.09). [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) have emerged as critical regulators of the progression of human cancers, including colorectal cancer (CRC). The study of genome-wide lncRNA expression patterns in metastatic CRC could provide novel mechanism underlying CRC carcinogenesis. In here, we determined the lncRNA expression profiles correlating to CRC with or without lymph node metastasis (LNM) based on microarray analysis. We found that 2439 lncRNAs and 1654 mRNAs were differentially expressed in metastatic CRC relative to primary CRC. Among these dysregulated lncRNAs, FBXL19-AS1 was the most significantly upregulated lncRNA in metastatic tumors. Functionally, knockdown of FBXL19-AS1 played tumor-suppressive effects by inhibiting cell proliferation, migration and invasion in vitro and tumor growth and metastasis in vivo . Overexpression of FBXL19-AS1 was markedly correlated with TNM stage and LNM in CRC. Bioinformatics analysis predicted that miR-203 was potentially targeted by FBXL19-AS1, which was confirmed by dual-luciferase reporter assay. Pearson's correlation analysis showed that miR-203 expression was negatively related to FBXL19-AS1 in tumor tissues. Finally, miR-203 inhibition abrogated the effect of FBXL19-AS1 knockdown on the proliferation and invasion of LoVo cells. Our results reveal the cancer-promoting effect of FBXL19-AS1, acting as a molecular sponge in negatively modulating miR-203, which might provide a new insight for understanding of CRC development. [ABSTRACT FROM AUTHOR]

Purpose: Long non-coding RNA (LncRNA) urothelial carcinoma-associated 1 (UCA1) is reported to be dysregulated in hepatocellular carcinoma (HCC) progression. However, the functions of UCA1 in HCC still need further study. The aim is to detect the role of UCA1 involving in HCC cells proliferation and invasion, and epithelial-mesenchymal transition (EMT). Methods: The quantitative real-time PCR was used to detect the UCA1 and miR-203 expression levels in 60 cases' HCC tissues and adjacent normal tissues. Western blotting analysis was performed to detect the EMT markers E-cadherin, Vimentin and transcription factor Snail1, Snail2 expression. Luciferase reporter assay, RNA immunoprecipitation (RIP) and pull-down assays were used to evaluate whether miR-203 was a target of UCA1. Results: Our results showed that UCA1 was markedly upregulated in HCC tissues and higher UCA1 expression in HCC was positively associated with tumor size, vascular invasion and American Joint Committee on Cancer (AJCC) stage ( P < 0.05). Furthermore, gain-of-function and loss-of-function analysis showed that UCA1 knockdown inhibited HCC cells proliferation and invasion in vitro and xenograft tumour growth in vivo. Moreover, UCA1 overexpression promoted cell epithelial-mesenchymal transition (EMT) in HCC via effectively sponging to miR-203 and thereby activating the expression of transcription factor Snail2. Conclusions: Our results identified that UCA1/miR-203/Snail2 pathway might involve in HCC progression. Inhibition of UCA1 acted as a promising therapeutic target for HCC patients. [ABSTRACT FROM AUTHOR]

Background: Recently experimental validation of the networks revealed bach1, a basic leucine zipper transcription factor, as the common regulator of several functional invasive genes. The expression of bach1 and its target genes was linked to the higher risk of breast cancer recurrence in patients. The aim of this study was to investigate the effect of specific bach1 small interfering RNAs, on the invasive and expression level of miR-203, miR-145, matrix metalloproteinase-9, and CXCR4 receptor which play a role in cancer metastasis, in MDA-MB-468 cell lines. Methods: Small interfering RNA transfection was performed with transfection regent. The survival effects of small interfering RNA were determined using trypan blue assay cells. The expression level of messenger RNA and matrix metalloproteinase-9 to assess cell invasion and the expression level of miR-203, miR-145, and CXCR4 receptor were measured by quantitative real-time polymerase chain reaction analysis on the MDA-MB-468 cell lines. Results: Transfection with small interfering RNA significantly suppressed the expression of bach1 gene in dose-dependent manner after 48 h (p < 0.0001). A significant reduction in cell invasion and CXCR4 receptor, matrix metalloproteinase-9 expression were observed (p < 0.0001). It was also a dramatic increase in the expression level of miR-203 and miR-145 (p < 0.0001). Conclusions: Our results suggest that the bach1-specific small interfering RNA effectively decrease CXCR4 receptor, matrix metalloproteinase-9 expression and breast adenocarcinoma cells invasive, also increased the expression of tumor-suppressive microRNA-203 and miR-145. Thus, these microRNAs may play a role in invasive/metastasis of carcinogenic breast cancer cells. Therefore, bach1 knockdown can be considered as a potent adjuvant in breast cancer therapy. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Cardiovascular disease is one of the diseases threatening human health. Myocardial fibrosis is a major cause of cardiovascular diseases. Studies have shown that over expression of miR-203 can inhibit the fibrosis. Therefore, in this study, the effect of differential expression of miR-203 on fibrosis of cultured mouse cardiomyocytes was investigated. MATERIALS AND METHODS: Activators and inhibitors of miR-203 were designed according to the sequence of miR-203, synthesized, and transfected into mouse cardiomyocytes to establish activator group, inhibitor group, and control group. The expression levels of fibrosis-related factors including FN, CTGF, and TGF-β1 were measured by Western blot and RT-PCR 24 h and 36 h after transfection. RESULTS: Over-expression of miR-203 in mouse cardiomyocytes significantly decreased the expression levels of TGF-β1, CTGF, and FN in a time-dependent manner, compared with that in the control group (p <0.05). Inhibition of miR-203 expression in mouse cardiomyocytes significantly increased the expression levels of TGF-β1, CTGF, and FN 36 h after transfection, compared with that in the control group (p < 0.05). No significant differences were seen in the expression levels of TGF-β1, CTGF, and FN 24 h after transfection, compared with that in the control group (p >0.05). CONCLUSIONS: Over-expression of miR-203 in mouse cardiomyocytes significantly decreased the expression levels of TGF-β1, CTGF, and FN, which might be used as a detection index for prediction of fibrosis. [ABSTRACT FROM AUTHOR]

Background/Aims: MicroRNA (miRNA) plays important roles in the development of different cancers. In this study, we investigated the roles and mechanisms of miR-203 in human cervical cancer. Methods: miR-203 expression was detected in cervical cancer tumors and cell lines by qRT-PCR. The methylation status in the promoter region of miR-203 was examined by methylation-specific PCR. The functional effect of miR-203 was determined by both in vitro and in vivo assays. Results: miR-203 was frequently down-regulated in cervical cancer tumors and cell lines. This down-regulation of miR-203 was associated with methylation of the miR-203 promoter. Furthermore, miR-203 down-regulated vascular endothelial growth factor alpha (VEGFA) expression by directly targeting its 3'-untranslated region. Functional assays revealed that miR-203 suppressed cervical cancer cell proliferation, tumor growth, and angiogenesis in nude mice, whereas forced expression of VEGFA rescued this inhibitory effect. Conclusion: Our collective findings indicate that miR-203 functions as a tumor suppressor by targeting VEGFA, resulting in the inhibition of tumor growth and angiogenesis. Thus, miR-203 may be a potential therapeutic target and prognostic marker in cervical cancer.

Problem To determine whether miR-203 mediates endothelial inflammatory response in preeclampsia. Method of study Maternal vessel miR-203 expression was assessed by in situ hybridization. Suppressor of cytokine signaling-3 ( SOCS-3) and ICAM expression was determined by immunostaining. Subcutaneous fat tissue sections from normal and preeclamptic pregnant women were used. miR-203-induced inflammatory response was evaluated by the measurements of IL-6, IL-8, ICAM, and VCAM expression and production and neutrophil adhesion in the endothelial cells ( EC) transfected with miR-203 precursor, pre-miR-203. SOCS3 expression was also determined. Results Up-regulation of miR-203 and ICAM expression and down-regulation of SOCS-3 expression were demonstrated in maternal vessel endothelium in preeclampsia. Overexpression of miR-203 resulted in down-regulation of SOCS-3 expression and increases in the production of IL-6, IL-8, ICAM, and VCAM and neutrophil adhesion in ECs. Conclusion As miR-203 is an inflammatory micro RNA, increased miR-203 production/expression in ECs could diminish an anti-inflammatory activity and increase the endothelial inflammatory response in preeclampsia. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) play critical roles in tumorigenesis and cancer metastasis. Recently, miR-203 was reported as a tumor suppressor microRNA silenced in different malignancies including hepatocellular carcinoma, prostate cancer, oral cancer, breast cancer, and hematopoietic malignancy, whereas its role in the carcinogenesis of gastric carcinoma (GC) has not been evaluated. Here, we analyzed the levels of miR-203 and Slug in the GC specimen and studied their correlation. We analyzed the binding of miR-203 to the 3′-UTR of Slug messenger RNA (mRNA) and its effects on Slug translation by bioinformatics analysis and by luciferase-reporter assay, respectively. We modified miR-203 levels in GC cells and studied their effects on the cell invasiveness in transwell cell migration assay. We found that in GC, miR-203 levels were significantly decreased and Slug levels were significantly increased. miR-203 and Slug inversely correlated in patients' specimen. Bioinformatic analysis predicted that miR-203 may target the 3′-UTR of Slug mRNA to inhibit its translation, which was confirmed by luciferase-reporter assay. Overexpression of miR-203 inhibited Slug and cell invasiveness, while depletion of miR-203 increased Slug and cell invasiveness. These data suggest that miR-203 suppression in GC promotes Slug-mediated cancer metastasis. [ABSTRACT FROM AUTHOR]

The purpose of this study was to explore the role of miR-203 in colorectal cancer (CRC) and evaluate the correlation between miR-203 and BIRC5. The expressions of miR-203 in the tissues of 122 CRC patients (with non-tumor tissues as controls) and those from 30 healthy donors were detected by TaqMan® MicroRNA assay. BIRC5s expressions in CRC and non-tumor tissues were detected by immunohistochemistry. Significantly less miR-203 was expressed in CRC tissues ( P < 0.05) than in non-tumor tissues. Furthermore, low expression level of miR-203 was correlated with distant metastasis (DM), lymph node metastasis (LNM), and TNM stage ( P < 0.05), but there were no significant differences between tumor size or gender. The positive expression rates of BIRC5 in CRC and non-tumor tissues were 73.77 % (90/122) and 30.32 % (37/122), respectively. The expression intensity of BIRC5 in CRC was significantly higher than that of non-tumor tissues ( P < 0.05). It was significantly correlated with DM, LNM, and TNM stage ( P < 0.05). Finally, miR-203 expression was negatively associated with that of BIRC5 ( r = −0.8150, P < 0.05). In conclusion, miR-203 was down-regulated in CRC tissues and involved in the onset and progression of CRC. The expressions of miR-203 and BIRC5 in CRC were significantly negatively correlated, suggesting that BIRC5 may be regulated by miR-203. miR-203 is a potential suppressor and predictive biomarker for CRC. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) play an important role in the tumorigenesis of ovarian cancer. Previously, we have reported the dysregulation of miR-203 in the ovarian cancer tissues. However, the biological functions and molecular mechanisms of miR-203 in ovarian cancer remain unknown. Here, we showed that the expression of miR-203 was increased in ovarian cancer tissues compared with the adjacent non-cancerous tissues and the transcription of miR-203 was inhibited by P53. Forced expression of miR-203 in ovarian cancer promoted cell growth and migration, while depletion of miR-203 inhibited the growth and migration of ovarian cancer cells. In addition, miR-203 promoted the metastasis of ovarian cancer cells in vivo and shorted the survival of the nude mice. Mechanically, miR-203 targeted the 3′-UTR of pyruvate dehydrogenase B (PDHB) and increased the consumption of glucose and the production of lactate. Overexpression of PDHB abolished the oncogenic effects of miR-203 on the growth of ovarian cancer cells. Together, our data suggested the oncogenic roles of miR-203 in ovarian cancer by promoting glycolysis, and miR-203 might be a therapeutic target for ovarian cancer. [ABSTRACT FROM AUTHOR]

Background: As the end product of the bacterial fermentation of dietary fiber in the colonic lumen, sodium butyrate (NaBt) has been reported to exert antitumor effects on colorectal cancer (CRC). In addition to functioning as a histone deacetylase (HDAC) inhibitor, NaBt also regulates the expression of microRNAs (miRNAs) to inhibit CRC cell proliferation. Yet, the mechanisms involved are not completely understood. Here we investigate whether NaBt regulates miR-203 to inhibit CRC growth and explore the promising target gene of miR-203 in CRC cells. Methods: We conducted qRT-PCR and Western blotting assays to evaluate the effects of NaBt on the expression of miR-203 and NEDD9 in HT-29 and Caco-2 cell lines. The promising target gene of miR-203 was predicted by miRNA target prediction and dual luciferase reporter assay. CRC Cell proliferation, colony formation, cell apoptosis and cell invasion assays were performed to explore the effect of NaBt, miR-203 and NEDD9 on HT-29 and Caco-2 cell lines. Results: The results showed that NaBt increased the expression of miR-203 to induce CRC cell apoptosis as well as inhibit cell proliferation, colony formation and cell invasion. Moreover, we determined that the NEDD9 was a target gene of miR-203. NEDD9 partially overcame the inhibitory effects of miR-203 on CRC cell colony formation and invasion. Conclusions: NaBt could induce CRC cell apoptosis, inhibit CRC cell proliferation, colony formation and invasion through miR-203/NEDD9 cascade. The present study may enrich the mechanisms underlying the process that NaBt exerts anti-tumor effects on CRC cells. [ABSTRACT FROM AUTHOR]

Breast cancer is the second leading cause of cancer mortality in women worldwide. Molecular therapy is needed to improve the outcome in patients with breast cancer. miR-203 participates in cancer cell proliferation, transformation, and apoptosis. This study showed that miR-203 was upregulated in breast cancer tissues and the MCF-7 cell line. miR-203 knockdown suppressed colony formation and transformation and also limited migration in MCF-7 cells. Fibroblast growth factor 2 (FGF2) was confirmed as a novel target of miR-203, as miR-203 knockdown induced an enhanced expression of FGF2 in MCF-7 cells. Moreover, FGF2 can reverse transforming growth factor-ß signal pathway to suppress breast cancer. These findings provide new insights with potential therapeutic applications for the treatment of breast cancer. [ABSTRACT FROM AUTHOR]

Background Atherosclerosis is a chronic immune-inflammatory disorder and one of the leading causes responsible for cardiovascular morbidity and mortality. Traditional Chinese medicine treatment with multi-targets has shown prospects for the therapeutic effect on atherosclerosis. Thus, this study aims to investigate whether xiaoxianggou has benefit for reducing the atherosclerotic plaque area in endogenous high Ang II ApoE −/− mice and investigated the underlying mechanisms. Methods Endogenous high Ang II ApoE −/− mice model was generated by using two kidney one clip (2K1C). All mice were treated by intragastric administration with xiaoxianggou two times a week for 16 weeks. En face plaque area was analyzed by oil-red O staining. Serum anti-OxLDL antibodies were measured by ELISA assay. Expression of miR-203 and Ets-2 were evaluated using qRT-RCR and western blotting analysis, respectively. Results This study revealed that xiaoxianggou treatment dose-dependently reduced the atherosclerotic plaque area and serum autoantibodies against oxLDL, elevated miR-203 expression and reduced Ets-2 expression in endogenous high Ang II ApoE −/− mice. In primary arterial ECs, Xiaoxianggou reverses the reduced miR-203 expression and the elevated Ets-2 expression induced by AngII, which was further recovered by miR-203 inhibitor. Additionally, miR-203 regulated the expression of Ets-2 by targeting Ets-2-3′ UTR. Moreover, miR-203 inhibitor reversed the reduction of atherosclerotic lesion area induced by Xiaoxianggou. Conclusions These findings present that xiaoxianggou plays an anti-atherosclerotic role in endogenous high Ang II ApoE −/− mice model, which is partly due to its antioxidant actions against atherosclerosis and the inhibition of miR-203 on the expression of Ets-2 in endothelial cells. [ABSTRACT FROM AUTHOR]

Background: Oral lichen planus (OLP) is a T cell-mediated autoimmune disease involving oral mucosa. Interleukin-22 (IL-22) as the signature cytokine of T helper 22 cells is increasingly recognized as a key regulator in various autoimmune diseases. Our previous study reported that IL-22 immunoexpression in OLP was significantly increased compared with the normal controls.Methods: The objective of this preliminary study was to compare the IL-22 expression levels in oral biopsies from patients with OLP (n = 50) against normal oral mucosa (n = 19) using RT-qPCR and Western blot, identify the potential targeting miRNAs of IL-22, and examine the miRNA expression levels in OLP.Results: Interleukin-22 expression level in OLP was significantly increased compared with the normal controls. The Dual-Luciferase reporter assay system in human embryonic kidney 293 (HEK293) cells demonstrated that miR-562 and miR-203 were the target miRNAs of IL-22, which was consistent with predictions from bioinformatics software analyses. Interestingly, miR-562 expression in OLP was significantly decreased, but miR-203 expression in OLP was significantly increased compared with the normal controls.Conclusion: This preliminary study for the first time reported that aberrant expression levels of miR-562 and miR-203 were associated with high expression of IL-22 and demonstrated the target relationship between miRNAs and IL-22 in HEK293 cells. Our data indicated that IL-22 and its targeting miRNAs contribute to the pathogenesis of OLP. Further studies are required to investigate the regulatory pathways of IL-22 and miR-562 and miR-203 in OLP. [ABSTRACT FROM AUTHOR]

Background: Metastasis is a major cause of death in patients with gastric cancer (GC). MicroRNAs (miRNAs) relating to the epithelial-mesenchymal transition (EMT) control GC progression and metastasis. The aim of this study was to evaluate serum EMT-associated miRNAs for metastatic and prognostic noninvasive biomarkers in GC. Methods: In the first step of this study (preliminary experiments), we selected candidate miRNAs associated with metastasis by analyzing the expression of the miR-200 family (miR-200a, miR-200b, miR-200c, miR-141, and miR-429) and miR-203 in serum samples from stage I ( n = 12) and stage IV ( n = 12) GC patients. The second phase involved the independent validation of candidate miRNAs in serum specimens from 130 patients with GC and 22 controls. Results: Based on the preliminary experiments, miR-203 was selected as the candidate serum miRNA that was most closely associated with metastasis. Validation analysis revealed that serum miR-203 levels were significantly lower in stage IV than stage I-III GC patients. Serum miR-203 expression was significantly lower in GC patients with a higher T stage, vessel invasion, and lymph node, peritoneal, and distant metastases. Low expression of serum miR-203 was significantly associated with poor disease-free and overall survival. Multivariate analysis revealed that low serum miR-203 expression was an independent predictive marker for lymph node, peritoneal, and distant metastases and a poor prognosis in patients with GC. Conclusions: Serum miR-203 has the potential to serve as a noninvasive biomarker for prognosis and to predict metastasis in patients with GC. [ABSTRACT FROM AUTHOR]