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A sensitive biosensor for the detection of miR-141 has been constructed. The DNA-biosensor is prepared by first immobilizing the thiolated methylene blue-labeled hairpin capture probe (MB-HCP) on two-layer nanocomposite film graphene oxide-chitosan@ polyvinylpyrrolidone-gold nanourchin modified glassy carbon electrode. We used the hematoxylin as an electrochemical auxiliary indicator in the second stage to recognize DNA hybridization via the square wave voltammetry (SWV) responses that record the accumulated hematoxylin on electrode surfaces. The morphology and chemical composition of nanocomposite was characterized using TEM, FE-SEM, and FT-IR techniques. The preparation stages of the DNA-biosensor were screened by electrochemical impedance spectroscopy and cyclic voltammetry. The proposed DNA-biosensor can distinguish miR-141 from a non-complementary and mismatch sequence. A detection limit of 0.94 fM and a linear range of 2.0 –5.0 × 105 fM were obtained using SWV for miR-141 detection. The working potential for methylene blue and hematoxylin was -0.28 and + 0.15 V vs. Ag/AgCl, respectively. The developed biosensor can be successfully used in the early detection of non-small cell lung cancer (NSCLC) by directly measuring miR-141 in human plasma samples. This novel DNA-biosensor is of promise in early sensitive clinical diagnosis of cancers with miR-141 as its biomarker. [ABSTRACT FROM AUTHOR]

Parkinson's disease is known as the second most common neurodegenerative disorder that commonly affects the older population. MicroRNAs are categorized as short non-coding RNAs (19–24 nt) involved in the post-transcriptional regulation of gene expression in multicellular organisms. They can influence both stability and translation of mRNAs. Recent studies suggested that miR-141 which is well-known in oxidative-stress pathway is up-regulated in some neurodegenerative diseases including the Alzheimer disease. This study was intended to evaluate the miR-141 expression and its predicted target gene in the MPP+ treated differentiated SH-SY5Y cells. In silico studies were also considered by using miRWalk and TargetScan databases to determinate any possible target of miR-141. Moreover, SHSY-5Y cells were allowed to be exposed to MPP+ as a parkinsonian neurotoxin. The quantification of the expression levels of miR-141 and the predicted target mRNA was done by using RT-qPCR to show their expression pattern. Results showed the significant up-regulation of miR-141 during the treatment of human SH-SY5Y cells with MPP+, inducing oxidative stress and apoptosis. Furthermore, transcript level of SIRT1 in these cells exhibited significant down-regulation under MPP+ treatment. In addition, exposure to MPP+ raised the apoptotic rate, leading to ROS accumulation and oxidative stress. To summarize, it was found that miR-141 expression could trigger the PD-related pathogenic processes and decreased SIRT1 expression is associated with increased miR-141 expression in MPP+ treated SHSY-5Y cells. [ABSTRACT FROM AUTHOR]

Purpose: Recent studies have demonstrated the differential expression of micro(mi)RNAs in endometriosis. Previously, we reported the low expression of miR-141 in patients with this disease. Epithelial-to-mesenchymal transition (EMT) and the transforming growth factor-beta1 (TGF-β1)-induced SMAD2 signalling pathway are central to tumour proliferation and invasion. However, the role of miR-141 in regulating the TGF-β1/SMAD2 signalling pathway and the associated EMT to be elucidated.Methods: The levels of TGF-β1/SMAD2 signalling and EMT markers expression in eutopic and ectopic endometria of endometriosis were determined by immunohistochemistry and western blot analyses. MiR-141 expression was analysed by quantitative reverse-transcription polymerase chain reaction. Cellular invasion and proliferation were determined by transwell and CCK-8 assays, respectively. Functional assay of miR-141 was performed using plasmid and shRNA transfection methods.Result: The presence of miR-141, EMT, and TGF-β1/SMAD2 signalling markers were detected in eutopic and ectopic endometria of endometriosis. TGF-β1-induced EMT in Ishikawa (ISK) cells by activating the SMAD2 signalling pathway, whereas miR-141 inhibited the TGF-β1-induced EMT, proliferation and invasion abilities of these cells.Conclusion: These data identify miR-141 as a novel driver of EMT in endometriosis, implicates the link between miR-141 and TGF-β1/SMAD2 signalling pathway in the context of endometriosis, and underscore the role of EMT in the development of endometriosis. [ABSTRACT FROM AUTHOR]

In the present study, we intend to determine whether Sestrin proteins 1, 2, and 3 (SESN1-3) are targets of microRNA-200 family (miR-200) in endometrial cancer (EC) Ishikawa, AN3CA, KLE, and RL 95-2 cell lines and to investigate how these potential interactions influence anoikis resistance of EC cell lines. The luciferase reporter assay, qRT-PCR, and western blotting assays were used to verify whether SESN1-3 are direct targets of miR-200. Moreover, the anoikis assay and transient transfections of miR-200 mimics or inhibitors into EC cell lines were performed to evaluate the modulatory role of miR-200 and SESN proteins on anoikis resistance. We demonstrated that SESN2 protein is a direct target of mir-141 in KLE and RL-95-2 EC cell lines and the functional interaction of miR-141 and SESN2 protein has a downstream effect on anoikis resistance and SESN2 expression level in Ishikawa and AN3CA cell lines. Moreover, we have shown that SESN3 protein is a direct target of miR-200b, miR-200c, and miR-429 in Ishikawa, AN3CA, and KLE cell lines. Our results show that manipulation of miR-200b, miR-200c, and miR-429 expression patterns also has an influence on anoikis resistance in EC cell lines. In conclusion, we identified new interactions between miR-200 and the oxidative stress response SESN proteins that affect anoikis resistance in human EC cells. [ABSTRACT FROM AUTHOR]

To understand the role of microRNA-141 (miR-141) in hypoxia/reoxygenation (H/R)-induced PC12 cell injury via modulation of Keap1/Nrf2 signaling pathway. PC12 cells were divided into Control, H/R, H/R + miR-141 mimics, H/R + NC, H/R + miR-141 inhibitor, H/R + siKeap1 and H/R + miR-141 inhibitors+siKeap1 groups. The expression of miR-141 and Keap1/Nrf2 pathway was measured by qRT-PCR and western blotting, cell viability evaluated by MTT assay while cell apoptosis tested by flow cytometry. Besides, MDA (malondialdehyde), SOD (Super Oxide Dismutase) and LDH (lactate dehydrogenase) levels were determined. DCFH-DA and JC-1 staining were used to measure ROS and mitochondrial membrane potential (MMP) respectively. Compared with Controls, PC12 cells induced by H/R exhibited decreased cell viability and increased cell apoptosis rate, with elevated MDA, LDH and ROS and reduced SOD levels; and meanwhile, MMP and miR-141 expression were declined, whereas cytoplasmic Nrf2 levels were enhanced with the downregulated nuclear Nrf2 level (all P < 0.05). However, these cells treated with miR-141 mimics and siKeap1 showed obvious improvement in H/R-induced cell injury, while miR-141 inhibitors presented significantly aggravated cell injury (both P < 0.05). Besides, siKeap1 can reverse the effect of miRNA-141 inhibitors on aggravating H/R-induced PC12 cell injury. miR-141-mediated Keap1/Nrf2 signaling pathway to promote cell viability, inhibit cell apoptosis and reduce oxidative stress of PC12 cells, thereby alleviating H/R-induced cell injury. [ABSTRACT FROM AUTHOR]

The miR-200 family, consisting of miR-200a/b/c, miR-141, and miR-429, is well known to inhibit epithelial-to-mesenchymal transition (EMT) in cancer invasion and metastasis. Among the miR-200 family members, miR-200a/b/c and miR-429 have been reported to inhibit angiogenesis. However, the role of miR-141 in angiogenesis remains elusive, as contradicting results have been found in different cancer types and tumor models. Particularly, the effect of miR-141 in vascular endothelial cells has not been defined. In this study, we used several in vitro and in vivo models to demonstrate that miR-141 in endothelial cells inhibits angiogenesis. Additional mechanistic studies showed that miR-141 suppresses angiogenesis through multiple targets, including NRP1, GAB1, CXCL12β, TGFβ2, and GATA6, and bioinformatics analysis indicated that miR-141 and its targets comprise a powerful and precise regulatory network to modulate angiogenesis. Taken together, these data not only demonstrate an anti-angiogenic effect of miR-141, further strengthening the critical role of miR-200 family in the process of angiogenesis, but also provides a valuable cancer therapeutic target to control both angiogenesis and EMT, two essential steps in tumor growth and metastasis. [ABSTRACT FROM AUTHOR]

Background: Prostate cancer (PCa) is a common malignant human tumor and one of the main causes of cancer-related deaths in men. At present, prostate-specific antigen levels are widely used to diagnose PCa in the clinic, but they are not sufficient for an accurate early diagnosis or prognosis.Methods: To identify potential molecular markers for PCa, we used real-time PCR to measure the expression levels of various microRNAs, including miR-1825, miR-484, miR-205, miR-141, and let-7b, in the serum of 72 PCa patients and 34 healthy controls.Results: miR-1825, miR-484, miR-205, miR-141, and let-7b were shown to be highly specific for PCa, suggesting that they could be used as PCa tumor screening biomarkers. miR-205 may also be used as a biomarker for indicating bone metastasis in PCa patients, miR-1825 levels may help indicate tumor-node-metastasis classification, the evaluation of treatment effects, and determining prognosis, while let-7b levels may indicate potential tumor malignancy and the hormone resistance status and could be used as a basis to adjust individual treatments for the high-risk, early diagnosis of refractory PCa.Conclusion: This study identified possible PCa tumor markers to more accurately predict the occurrence, progression, and prognosis of PCa, and which could be used in the development of tumor drug therapy. [ABSTRACT FROM AUTHOR]

Chronic prostatitis was the most common type of prostatitis and oxidative stress was reported to be highly elevated in prostatitis patients. In this study, we determined the effect of N-acetylcysteine (NAC) on prostatitis and the molecular mechanism involved in it. Male Sprague-Dawley rats were divided into three groups: control group (group A, n = 20), carrageenan-induced chronic nonbacterial prostatitis (CNP) model group (group B, n = 20), and carrageenan-induced CNP model group with NAC injection (group C, n = 20). Eye score, locomotion score, inflammatory cell count, cyclooxygenase 2 (COX2) expression, and Evans blue were compared in these three groups. The expression of miR-141 was determined by quantitative real-time PCR (qRT-PCR). Moreover, protein expressions of Kelch-like ECH-associated protein-1 (Keap1) and nuclear factor erythroid-2 related factor 2 (Nrf2) and its target genes were examined by Western blot. Luciferase reporter assay was performed in RWPE-1 cells transfected miR-141 mimic or inhibitor and the plasmid carrying 3′-UTR of Keap1. The value of eye score, locomotion score, inflammatory cell count, and Evans blue were significantly decreased in group C, as well as the expression of COX2, when comparing to that of group B. These results indicated that NAC relieved the carrageenan-induced CNP. Further, we found that NAC increased the expression of miR-141 and activated the Keap1/Nrf2 signaling. Luciferase reporter assay revealed that miR-141 mimic could suppress the activity of Keap1 and stimulate the downstream target genes of Nrf2. In addition, miR-141 inhibitor could reduce the effect of NAC on prostatitis. NAC ameliorates the carrageenan-induced prostatitis and prostate inflammation pain through miR-141 regulating Keap1/Nrf2 signaling. [ABSTRACT FROM AUTHOR]

Background: MiR-141 and long noncoding RNA MEG3 have been independently reported to be tumor suppressor genes in various cancers. However, their expression has never been previously associated with gastric cancer (GC).Aims: To investigate the interaction of miR-141 and MEG3 in GC.Methods: QRT-PCR was used to detect miR-141, MEG3, and E2F3 in gastric tissues and cells. CCK-8 and flow cytometry analysis were used to detect cell functions. Western blot and luciferase activity were used to identify E2F3 as one of the direct targets of miR-141.Results: We found that expression of both miR-141 and MEG3 was significantly reduced in GC compared with levels in matched nonmalignant tissues. Positive correlation between miR-141 and MEG3 was found in both tumor tissues and control tissues. Furthermore, the over-expression of either miR-141 or MEG3 in 7901 and MKN45 cells inhibited cell proliferation and cell cycle progression and promoted cell apoptosis. E2F3 was identified as a target of miR-141, and its inhibition significantly reduced MEG3 expression. E2F3 expression was also found to be negatively associated with both MEG3 and miR-141. E2F3 over-expression partly reversed the changes caused by transfection of miR-141 mimic, and inhibition of miR-141 or MEG3 overrides MEG3- or miR-141-induced modulation of cell growth in GC.Conclusions: These findings together suggested that miR-141 could be interacting with MEG3 and targeting E2F3, and these factors may play important anti-tumor effects in GC pathogenesis and provide therapeutic targets in the clinics. [ABSTRACT FROM AUTHOR]

Ovarian cancer is the fifth leading cause of female death globally due to its low survival rates. Thus, improved approaches for ovarian cancer detection are urgently needed. MicroRNAs as a new class of biomarkers have been explored in recent studies. This study was trying to identify and validate the two kinds of serum microRNA (miR-200c and miR-141) as biomarkers for ovarian cancer. We extracted serum samples from 74 epithelial ovarian cancer patients, 19 borderline ovarian cancer, and 50 healthy controls. Relative expression of these miRNA markers were measured by quantificational real-time polymerase chain reaction assay (qRT-PCR). Receiver operating characteristics (ROC) and area under the ROC curve (AUC) were used to validate the diagnostic value of miR-200c and miR-141. Kaplan-Meier curve and the log-rank test were conducted to detect the prognostic value of miR-200c and miR-141. miR-200c and miR-141 were significantly elevated in the epithelial ovarian cancer patients compared to healthy controls. The relative expression level of miR-200c showed a descending trend from early stages to advanced stages, while the level of miR-141 displayed an escalating trend. Patients with high miR-200c level achieved significantly a higher 2-year survival rate compared with the other group ( P < 0.001), while low miR-141 group showed a significantly higher survival rate. The results of the current study suggested that serum miR-200c and miR-141 were able to discriminate the ovarian cancer patients from healthy controls. In addition, miR-200c and miR-141 may be predictive biomarkers for ovarian cancer prognosis. Further large-scale studies are still needed to confirm our findings. [ABSTRACT FROM AUTHOR]

The aims of this study are to analyze the association of microRNA-141 (miR-141) with the clinicopathological parameters of bladder cancer and evaluate the value of miR-141 in predicting the prognosis of bladder cancer. In this study, 114 patients with bladder cancer were enrolled in the study, and tissue specimens were obtained from the tumor zone and from adjacent normal area. miR-141 expression was determined using SYRB Green quantitative real-time polymerase chain reaction assay and was further correlated with patients' clinicopathological parameters and the follow-up data. The results indicated that miR-141 was upregulated in malignant bladder specimens compared with normal ones ( P < 0.001). miR-141 expression was significantly associated with tumor stage ( P < 0.001), tumor grade ( P < 0.001), and muscle invasion status ( P < 0.001). Log-rank test showed that the higher miR-141 expression was associated with longer disease-specific survival of the patients with bladder cancer ( P < 0.001), which was also proven by univariate and multivariate Cox regression analysis ( P < 0.001 and P = 0.039, respectively). Focusing on patients with non-muscle invasive bladder cancer, univariate analysis using log-rank test and Cox regression analysis found that patients with high miR-141 expression experienced longer disease-free survival ( P = 0.031 and P = 0.040, respectively) and disease-specific survival ( P = 0.028 and P = 0.038, respectively), which was confirmed by multivariate Cox regression analysis ( P = 0.036 and P = 0.042, respectively). In conclusion, this study showed that miR-141 may contribute to the progression of bladder cancer and its upregulation may be independently associated with favorable prognosis of bladder cancer, suggesting that miR-141 might serve as a promising biological marker for further risk stratification in the management of bladder cancer. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) function as essential post-transcriptional modulators of gene expression involved in a wide range of physiologic and pathologic states, including cancer. Numerous miRNAs have been deregulated in hepatocellular carcinoma (HCC). Here, we investigated the role of miR-141 in HCC. Decreased expression of miR-141 was observed in both HCC tissues and cell lines. Ectopic overexpression of miR-141 reduced proliferation, migration, and invasion of HCC cells. E2F transcription factor 3 (E2F3) was confirmed to be a target of miR-141 in HCC cells. Moreover, restoration of E2F3 significantly reversed the tumor suppressive effects of miR-141. Our results suggest a critical role of miR-141 in suppressing metastasis of HCC cells by targeting E2F3. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. Increasing evidence suggests that microRNAs (miRNAs) are associated with HCC tumorigenesis. The present study was designed to define the role of miR-141 in HCC. The expression of miR-141 was significantly decreased in four HCC cell lines. Overexpression of miR-141 suppressed both the growth and the motility of HCC cells. Furthermore, we identified zinc finger E-box binding homeobox 2 (ZEB2) as a target of miR-141 and miR-141 functioned as a tumor suppressor via ZEB2 targeting in HCC. These data provide a novel potential therapeutic target for HCC treatment. [ABSTRACT FROM AUTHOR]

Osteosarcoma is the most common primary malignancy to arise from bone. The pathogenesis of osteosarcoma is unclear, and new therapy molecular target is needed. The miRNAs researched suggested that miRNAs are involved in the pathogenesis of osteosarcoma. MiR-141, which belong to miR-200 family, take a part in tumorigenesis. However, the role of miR-141 in the pathogenesis of osteosarcoma remained unclear. In this study, we focused on the miR-141 in osteosarcoma and found that the expression of miR-141 is lower in osteosarcoma. Overexpression of miR-141 not only inhibits osteosarcoma cell proliferation but also induces cell apoptosis. It is estimated that miR-141 played its role via ZEB1 and ZEB2. In all, miR-141 played a osteosarcoma-suppressing role via ZEB1 and ZEB2. Our finding may elucidate the miRNAs mechanism in osteosarcoma and provide a new molecule target for osteosarcoma therapy. [ABSTRACT FROM AUTHOR]

Purpose: Recent studies indicate that circulating microRNAs in serum/plasma are a novel class of non-invasive biomarkers with diagnostic and prognostic information. So far, circulating microRNAs have not been analyzed in patients with bladder cancer. Methods: We collected serum from patients with non-muscle invasive bladder cancer (NMIBC), muscle invasive bladder cancer (MIBC) and non-malignant urological disease. Total RNA was isolated from 400 μl of serum using the mirVana PARIS Kit; the artificial cel-miR-39 was spiked-in prior to RNA isolation to control different RNA isolation efficiencies. Quantitative real-time PCR was applied to measure the levels of 22 microRNAs upregulated in BCA tissue (miR-15a, miR-18a, miR-21, miR-93, miR-96, miR-103, miR-130b, miR-135b, miR-141, miR-182, miR-183, miR-190, miR-191, miR-200b, miR-422b, miR-425, miR-449b, miR-601, miR-639, miR-644, miR-649 and miR-1233) in the marker identification cohort (NMIBC, n = 11, MIBC, n = 10; controls, n = 10). The most promising serum microRNAs were tested in a validation cohort (NMIBC, n = 65, MIBC, n = 61; controls, n = 105). Results: The RNA recovery was similar in patients with NMIBC, MIBC and control subjects. The analysis of serum microRNA levels in the marker identification cohort indicated that serum miR-141 and miR-639 levels were increased in bladder cancer patients compared to CTRL. The analysis of these miR-141 and miR-639 in the validation cohort demonstrated that microRNA levels were similar in bladder cancer patients and control subjects. Furthermore, microRNA levels were not correlated with clinicopathological parameters (pT-stage, metastasis, grading). Conclusions: The analysis of serum miR-141 and miR-639 levels does not seem to be helpful in the diagnosis or prognosis of BCA. [ABSTRACT FROM AUTHOR]

miR-141 belongs to the miR-200 family, and has been found to be associated with numerous human malignancies; however, its role in gastric cancer (GC) has not been examined in detail. Here, we validated that miR-141 was decreased in GC tissues and cell lines. Forced expression of miR-141 significantly repressed GC cell proliferation and colony formation. Furthermore, miR-141 suppressed in vitro migration and invasion of GC cells. Hepatoma-derived growth factor (HDGF) was confirmed to be a direct target of miR-141 in GC cells. The suppressive effects of miR-141 on GC cell proliferation, colony formation, in vitro migration, and invasion were partially mediated by suppressing HDGF expression. Moreover, the expression of HDGF was negatively correlated with miR-141 in GC tissues. Our data suggest that miR-141 might be associated and plays essential role in GC progression. [ABSTRACT FROM AUTHOR]

Background: Colorectal cancer (CRC) is the third most common cancer in the world. Despite recent advances in diagnostics and treatment, prognosis for patients with advanced disease is still poor. microRNAs (miRNAs) are a class of endogenous, small noncoding RNA molecules which are crucial regulators of gene expression at the posttranscriptional level. miRNAs participate in many biological events and play an important role in various human diseases, including CRC.Aims: This study is to identify the role of miR-141 in migration and invasion of CRC cells.Methods: Expression of miR-141 and Smad interacting protein 1 (SIP1) were detected by real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western blotting. The effect of miR-141 on migration and invasion of CRC cells was investigated using wound healing assay and Matrigel invasion assay in vitro. The regulation effect of miR-141 on SIP1 was evaluated by dual-luciferase reporter assay.Results: We demonstrated that miR-141 levels correlate inversely with SIP1 protein levels as well as cell migration and invasion of CRC cells. SIP1 was identified as a functional target of miR-141.Conclusions: miR-141 regulates SIP1 to inhibit migration and invasion of CRC cells. miR-141 and SIP1 might be candidate therapeutic targets in CRC. [ABSTRACT FROM AUTHOR]

Human microRNA-141 (miR-141), a member of the miR-200 family, has been reported to be associated with various human malignancies. However, it remains unknown whether miR-141 is involved in the pathogenesis of gastric cancer. Therefore, we examined the expression of miR-141 in gastric cancer tissues and the effect of miR-141 overexpression on cancer cell proliferation. The expression level of miR-141 in 35 pair-matched gastric neoplastic and adjacent non-neoplastic tissues, and in 5 gastric cancer cell lines were examined by quantitative real-time PCR. The growth of MGC-803 cells transfected with miRNA precursor was examined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide) assay. MiR-141 was significantly down-regulated in 80% (28/35) of primary gastric cancer tissues compared with pair-matched adjacent non-tumor tissues ( P < 0.01). The expression of miR-141 was also found to be substantially reduced in several human gastric cancer cell lines such as MGC-803, HGC-27, SGC-7901 and BGC-823 cells. Overexpression of miR-141 with its precursors significantly inhibited the proliferation of gastric cancer cells. These results suggest that miR-141 may be involved in the development of gastric cancer through its inhibitory effect on cell proliferation. [ABSTRACT FROM AUTHOR]