Expression Levels Of miR-200a Research Articles

Reduction of cryopreservation-induced structural, functional and molecular damages in ram sperm by hydrated C60 fullerene.

This study aimed to investigate the morphological, functional and molecular changes in frozen-thawed ram sperm using an extender containing different concentrations of hydrated carbon 60 fullerene (C60 HyFn), a nanotechnological product. Semen taken from each of the seven Akkaraman rams were pooled. Semen collection was done twice a week and it continued for 3 weeks. Each pooled semen sample was divided into six equal groups and diluted with tris + egg yolk extender including 0 (control), 200, 400, 800 nM, 1 and 5 μM concentrations of C60 HyFn at 37°C. They were then frozen in liquid nitrogen vapour at -140°C, stored in liquid nitrogen container (-196°C) and thawed at 37°C for 25 s before analysis. In comparison with control, C60 HyFn addition prior to freezing procedure provided significant increases in total and progressive motility rates, glutathione peroxidase, catalase activities and percentage of highly active mitochondria, and significant decreases in dead and abnormal sperm rates, lipid peroxidation, caspase-3 and DNA fragmentation levels in frozen-thawed ram semen. When compared to control, C60 HyFn supplementation significantly down-regulated the expression levels of miR-200a and KCNJ11, and significantly up-regulated the expression levels of miR-3958-3p (at the concentrations of 200, 400, 800 nM and 1 μM), CatSper1 (at the concentrations of 200, 400 nM and 5 μM), CatSper2 (at the concentrations of 1 and 5 μM), CatSper3 (at the concentrations of 200, 400 nM, 1 and 5 μM), CatSper4 (at all concentrations), ANO1 (at the concentrations of 800 nM, 1 and 5 μM) and TRPV5 (at the concentrations of 200, 400 and 800 nM). The addition of C60 HyFn had no effect on global DNA methylation rates. As a result, C60 HyFn supplementation to ram semen extenders may be beneficial in reducing some of the functional, structural and molecular damages in sperm induced by the freeze-thawing procedure.

MicroRNA expression as a diagnostic parameter in early endometrial cancer

ObjectivesMicroRNAs (miRNAs) have emerged as biomarkers that showed strong diagnostic potential in various diseases, including cancer. This study aimed to estimate the expression and diagnostic potential of miRNAs (miR-200a, miR-21,...

High Expression of MicroRNA-200a/b Indicates Potential Diagnostic and Prognostic Biomarkers in Epithelial Ovarian Cancer.

Objective To detect the expression levels of microRNA-200a/b (miR-200a/b) in tumor tissues and serum of patients with epithelial ovarian cancer (EOC) and to explore its clinical significance. Methods A retrospective selection of 30 cases of benign ovarian disease or healthy physical examination (control group) and 55 cases of EOC patients. Real-time quantitative PCR was used to detect the expression level of miR-200a/b in tumor tissues and serum, and the miR-200a/b expresses relevance in the two types of samples were evaluated at the same time. Receiver operating characteristic curve (ROC) and Kaplan-Meier survival analysis were used to evaluate the diagnostic value of miR-200a/b expression and its influence on prognosis, respectively. Results The serum and tissue miR-200a/b expression levels in EOC patients were higher than those in the control group (P < 0.001), and there was a significant positive correlation between serum and tissue miR-200a/b expression (R2 = 0.9419, P < 0.001 and R2 = 0.9605, P < 0.001). ROC analysis showed that the expression of serum miR-200a/b can distinguish EOC patients from the control group. In addition, there were significant differences in the TNM stage, tumor differentiation, and lymph node metastasis between the miR-200a/b high- and low-expression groups (P < 0.05). Kaplan-Meier survival analysis found that the overall survival and disease-free survival of patients with high miR-200a/b expression were shorter than those of patients with low miR-200a/b expression (P < 0.05). Conclusion Upregulation of miR-200a/b expression is a common molecular event in EOC patients, and miR-200a/b can be used as a noninvasive biomarker for the diagnosis and prognosis of EOC.

Mir200a and miR4695-5p profiling as potential biomarkers for preterm birth

BackgroundPreterm labor is one of the main health problems and after birth congenital anomalies, the main cause of infant mortality. Epigenetic dysregulation and miRNA modulation have been indicated in preterm birth. Therefore, this study aimed to evaluate the miR200a and miR4695-5p profiling as potential biomarkers for preterm birth. Materials and methodsSerum samples were collected from a total of 90 patients (49 women with preterm labor and 41 women with term labor). After miRNA extraction, expression levels of miR200a and miR4695-5p were evaluated using RT-PCR. ResultsAccording to the results, the expression levels of miR200a and miR4695-5p were significantly different between the two groups (p-value = 0.015 and p-value = 0.031, respectively). Moreover, the expression of miR4695-5p was positively correlated with the expression of miR200a. However, considering the sex of the fetus, no significant difference was observed in the expression levels of miR200a and miR4695-5p in women with preterm or term labor carrying a female infant (p-value = 0.925). AUC-ROC analysis showed an acceptable sensitivity (60%) and specificity (75%) of miR4695-5p as a biomarker for preterm birth outcome (p-value = 0.031), suggesting this gene as a suitable biomarker for preterm birth. On the other hand, according to the sensitivity (73%) and specificity (51%) according to the AUC-ROC analysis for miR200a (p-value = 0.232) further studies are required to assure the efficacy of this microRNA as a biomarker of preterm birth. ConclusionIn overall, these microRNAs may be used as effective biomarkers for identifying women at risk of preterm labor for taking suitable preventive or treatment measures.

MicroRNA 200a inhibits liver fibrosis of schistosoma

ABSTRACT MicroRNA 200a (miR-200a) can inhibit the activation and proliferation of hepatic stellate cells (HSCs) through the transforming growth factor-β (TGF-β) signaling pathway, and improve fibrotic lesions. However, to date, there is no study exploring the role of miR-200a in schistosomiasis liver fibrosis (SLF). In this study, 64 healthy female Balb/c mice were selected and randomly divided into four groups: normal control group (non-infected schistosomiasis group), schistosomiasis model group, Lenti-NC group (lentivirus-negative control group), and Lenti-miR-200a group (lentivirus experimental group). Fluorescence quantitative PCR detection was used to measure the expression level of RNA. HE and Masson staining were used to observe the pathological changes of mouse liver tissue. Furthermore, ELISA was used to detect the serum concentrations of inflammation factors. We found that the expression level of miR-200a in liver tissues gradually decreased with the development of SLF. However, fibrosis factors (α-SMA and TGF-β2) and inflammatory cytokines (IL-4 and IFN-γ) in liver tissues and serum increased and the expression level of Colla I reached its peak in the 6th week of infection. Besides, compared with the schistosomiasis group and Lenti-NC group, the Lenti-NC group had lower levels of α-SMA, TGF-β2 and Colla I (P > 0.05). Furthermore, inflammatory cells and blue collagen fibers appeared and they increased with the development of infection in the schistosomiasis group and Lenti-NC group, but these changes reduced significantly in Lenti-miR-200a group. Our study demonstrated that upregulation of miR-200a might contribute to inhibiting schistosomiasis liver fibrosis.

Functional Link between miR-200a and ELK3 Regulates the Metastatic Nature of Breast Cancer.

Triple-negative breast cancer (TNBC) refers to breast cancer that does not have receptors for estrogen, progesterone, and HER2 protein. TNBC accounts for 10–20% of all cases of breast cancers and is characterized by its metastatic aggressiveness, poor prognosis, and limited treatment options. Here, we show that the metastatic nature of TNBC is critically regulated by a functional link between miR-200a and the transcription factor ELK3. We found that the expression levels of miR-200a and the ELK3 mRNA were negatively correlated in the luminal and TNBC subtypes of breast cancer cells. In vitro experiments revealed that miR-200a directly targets the 3’ untranslated region (UTR) of the ELK3 mRNA to destabilize the transcripts. Furthermore, ectopic expression of miR-200a impaired the migration and invasion of TNBC cells by reducing the expression level of the ELK3 mRNA. In in vivo studies, transfection of MDA-MB 231 cells (a claudin-low TNBC cell type) with exogenous miR-200a reduced their extravasation into the lung during 48 h after tail vein injection, and co-transfection of the cells with an expression plasmid harboring ELK3 that lacked an intact 3’UTR recovered their extravasation ability. Overall, our findings provide evidences that miR-200a and ELK3 is functionally linked to regulate invasive characteristics of breast cancers.

Enteropathogenic Escherichia coli (EPEC) infection inhibits intestinal vitamin C transporter function and expression: in vitro and in vivo studies

Enteropathogenic Escherichia coli (EPEC) is a food‐borne pathogen and causes enteric illnesses in children. EPEC infection is often associated with malnutrition of the host, especially in individuals experiencing severe and prolonged infection, and/or in those who are nutritionally compromised. While EPEC infection impacts nutrient transporter function and expression in intestinal epithelial cells, to date the consequences of EPEC infection on intestinal absorption of ascorbic acid (AA) are not known. Therefore, we investigated this issue using human intestinal epithelial Caco‐2 cells (in vitro) and mouse (in vivo) models. Infecting Caco‐2 cells with EPEC (WT) markedly inhibited AA uptake. EPEC mutants (ΔescN, ΔespA, ΔespB and ΔespD) did not affect AA uptake, whereas ΔespF and ΔespG/G2 mutants significantly inhibited AA uptake in Caco‐2 cells. Further, we observed significantly reduced expression of both hSVCT1 and hSVCT2 protein and mRNA levels upon infection. Similarly, infecting mice with EPEC (WT) significantly inhibited AA uptake and also reduced the expression of both mSVCT1 and mSVCT2 protein and mRNA levels in jejunum and colon. Studies have shown that epigenetic mechanisms (microRNA) play a role in mediating responses to bacterial infection in the intestine. Therefore, we determined miR103a, miR141 and miR200a expression levels, which were recently shown to regulate SVCT1 and SVCT2. Data from in vitro and in vivo models showed that all three microRNAs were up‐regulated significantly upon EPEC infection. In addition, expression of the accessory protein human glyoxalate reductase/hydroxypyruvate reductase (hGRHPR), which regulates the SVCT1 function, was also markedly decreased by EPEC in both models. These findings suggest that in intestinal epithelial cells, EPEC infection markedly inhibits AA uptake via dysregulation of both SVCT1 and SVCT2 expression.Support or Funding InformationSupported by NIH grants DK 107474, DK 58057, DK 56057, MH108154, GM088790, and a grant from the DVA

Gli3 is a novel downstream target of miR‑200a with an anti‑fibrotic role for progression of liver fibrosis invivo and invitro.

GLI family zinc finger 3 (Gli3), as the upstream transcriptional activator of hedgehog signaling, has previously been demonstrated to participate in the process of liver fibrosis. The present study aimed to investigate the potential functions of microRNA (miR)-200a and Gli3 in the progression of liver fibrosis. The expression levels of miR-200a and Gli3 in cells and tissues were determined by PCR and western blotting; the interaction of Gli3 and miR-200a was evaluated by bioinformatics analysis and dual-luciferase reporter assay. miR-200a was significantly reduced in serum samples from clinical patients, liver tissues of a carbon tetrachloride (CCl4)-induced rat model and activated LX2 cells. The expression of α-smooth muscle actin (α-SMA) and albumin at the mRNA and protein levels was increased and decreased in LX2 cells, respectively. However, the expression levels of α-SMA and albumin were reversed and Gli3 expression was markedly decreased in LX2 cells when transfected with miR-200a mimics. In addition, the dual-luciferase reporter assay confirmed the target interaction between miR-200a and Gli3. Finally, following the administration of miR-200a mimics to CCl4-induced rats, it was revealed that the alterations of α-SMA, albumin and Gli3 presented a similar trend to that in LX2 cells with miR-200a mimics transfection. Taken together, these results indicated that downregulation of miR-200a might enhance the formation of liver fibrosis, probably by targeting Gli3, and elevated miR-200a may attenuate the progression of liver fibrosis by suppressing Gli3. These findings suggested that miR-200a may function as a novel anti-fibrotic agent in liver fibrosis via inhibition of the expression of Gli3.

Helicobacter pylori Eradication Can Reverse the Methylation-Associated Regulation of miR-200a/b in Gastric Carcinogenesis.

Background/AimsEpigenetic change is one of the mechanisms that regulates the expression of microRNAs (miRNAs) and is known to play a role in Helicobacter pylori-associated gastric carcinogenesis. We aimed to evaluate the epigenetic changes of miR-200a/b in H. pylori-associated gastric carcinogenesis and restoration after eradication.MethodsThe expression and methylation levels of miR-200a/b were evaluated in gastric cancer (GC) cell lines, human gastric mucosa of H. pylori-negative and -positive controls, and H. pylori-positive GC patients. Next, the changes in the expression and methylation levels of miR-200a/b were compared between H. pylori-eradication and H. pylori-persistence groups at 6 months. Real-time reverse transcription-polymerase chain reaction was conducted to investigate the miRNA expression levels, and MethyLight was performed to assess the methylation levels.ResultsIn the GC cell lines, the level of miR-200a/b methylation decreased and the level of expression increased after demethylation. In the human gastric mucosa, the miR-200a/b methylation levels increased in the following group order: H. pylori-negative control group, H. pylori-positive control group, and H. pylori-positive GC group. Conversely, the miR-200a/b expression levels decreased in the same order. In the H. pylori-persistence group, no significant changes were observed in the methylation and expression levels of miR-200a/b after 6 months, whereas the level of methylation decreased and the level of expression of miR-200a/b increased significantly 6 months in the H. pylori-eradication group.ConclusionsEpigenetic alterations of miR-200a/b may be implicated in H. pylori-induced gastric carcinogenesis. This field defect for cancerization is suggested to be improved by H. pylori eradication.

MiR-200a improves respiratory distress syndrome in newborn rabbits via the Wnt/β-catenin signaling pathway.

The aim of this study was to investigate the effect of micro-ribonucleic acid (miR)-200a on respiratory distress syndrome (RDS) in newborn rabbits by regulating the Wnt/β-catenin signaling pathway. In this work, newborn rabbits aged three days were selected from our laboratory as research objects. The messenger RNA (mRNA) and protein expression levels of miR-200a, β-catenin and interleukin-10 (IL-10) in blood samples of healthy newborn rabbits and newborn rabbits with RDS were determined by fluorescence quantitative Polymerase Chain Reaction (PCR) and Western blotting, respectively. Lentivirus-packaged plasmids containing miR-200a were then injected into newborn rabbits suffering from RDS. After 2 d, the mRNA and protein expression levels of miR-200a, β-catenin and IL-10 in blood samples of newborn rabbits in different treatment groups were measured. Meanwhile, lung sections were collected from newborn rabbits in different treatment groups. After that, the sections were observed via hematoxylin and eosin (H&E) staining. At the same time, lung coefficient of newborn rabbits in different treatment groups was also measured. Compared with healthy newborn rabbits, the mRNA and protein expression levels of miR-200a and IL-10 in the blood of newborn rabbits with RDS decreased significantly (p<0.05), while β-catenin increased markedly (p<0.05). The mRNA and protein expression levels of β-catenin and IL-10 in newborn RDS rabbits with miR-200a over-expression and knockout were detected as well. The results revealed that lowly expressed miR-200a could remarkably promote the expression level of β-catenin, whereas inhibiting the expression of IL-10. However, highly expressed miR-200a could significantly inhibit the expression level of β-catenin and promote the expression level of IL-10. H&E staining results manifested that miR-200a knockout markedly promoted the increase of pulmonary alveoli with increased lung coefficients. However, the up-regulation of miR-200a could reduce lung coefficients and remarkably improve RDS. MiR-200a regulates RDS in newborn rabbits by regulating the Wnt/β-catenin signaling pathway.

Circulating epithelial-mesenchymal transition-associated miRNAs are promising biomarkers in ovarian cancer

Ovarian cancer is the fifth most common cause of cancer death among women that is mostly due to the difficulty of early diagnosis. Circulating miRNAs proved to be reliable biomarkers in various cancers. We screened 9 miRNAs, which are involved in epithelial-mesenchymal transition, in the plasma samples of patients with malignant (n = 28) or non-malignant (n = 12) ovarian tumors and disease-free healthy volunteers (n = 60) by qRT-PCR. The expression levels of miR200a, miR200b, miR200c, miR141, miR429, miR203a, miR34b (p < 0.001) and miR34a (p < 0.01) were significantly higher in the malignant samples than in healthy controls. MiR203a, miR141 (p < 0.01), miR200a and miR429 (p < 0.05) levels were also higher in malignant compared to non-malignant samples. ROC-AUC was the highest in the case of miR200c: 0.861 (95%CI = 0.776-0.947). Spearman's rank correlation analysis revealed positive correlation between the plasma levels of the studied miRNAs that was the highest between miR200b and miR200c (rs = 0.774; p < 0.001). Target analysis also suggested tight interaction between these miRNAs in the regulation of cancer development. The agreement of diagnostic tests based on miRNA levels and the standard CA125 or HE4 was weak according to Cohen's kappa values. We conclude that miR200 family members, miR34b and miR203a might be promising complementary biomarkers in ovarian cancer.

TCTP promotes epithelial-mesenchymal transition in lung adenocarcinoma.

BackgroundLung cancer is the most common and lethal malignancy worldwide. TCTP is highly expressed in various cancers including lung cancer. Epithelial–mesenchymal transition (EMT) could increase cancer cell invasion. Whether TCTP’s expression is associated with EMT in lung adenocarcinoma is largely unknown.MethodsSeveral Gene Expression Omnibus datasets were used to analyze the correlation between TCTP expression and overall survival of lung adenocarcinoma patients by Kaplan–Meier survival analysis. Then, 24 surgically removed fresh lung adenocarcinoma tissue samples and paired paracancer tissue samples were used to analyze the correlation between TCTP expression and tumor stage by immunohistochemical analysis. Furthermore, stable cell lines were generated using lentiviral transduction systems to knock down or overexpress TCTP in A549 cells. Cell migration and invasion were measured by scratch and transwell assays, and EMT marker proteins such as α-SMA, ZEB1, and E-cadherin were quantitated by Western blot. The expression levels of miR-200a, miR-141, and miR-429 were determined by real-time quantitative PCR, and their target genes were predicted by an online database miRTarBase. The interaction between TCTP and these genes was analyzed by String database and visualized by Cytoscape.ResultsTCTP was highly expressed in tumor tissues compared to paracancer tissues. The expression of TCTP was associated with shorter overall survival. TCTP knockdown experiment in A549 cells suggested that TCTP knockdown could decrease the migration and invasion of lung cancer cells, and the expression level of ZEB1 and α-SMA, but increase the expression of E-cadherin and p53. Vice versa, overexpression of TCTP could increase the migration and invasion of cancer cells, and the expression level of ZEB1 and α-SMA, but decrease the expression of E-cadherin and p53. Furthermore, we found the expression of miR-200a, miR-141, and miR-429 was associated with TCTP expression.ConclusionTCTP promotes EMT in lung adenocarcinoma, and this effect may be associated with miR-200 family members like miR-200a, miR-141, and miR-429.

MicroRNA-200 and microRNA-30 family as prognostic molecular signatures in ovarian cancer: A meta-analysis.

MicroRNAs (miRs) play a vital role in the occurrence, development, and progression of human cancers, but its role in the prognosis of ovarian cancer is unclear. We performed a meta-analysis by searching PubMed, Embase, and Web of Science databases for eligible studies. The pooled hazard ratios (HRs) and 95% confidence intervals (CIs) were used to explore the association between miRs expression and overall survival (OS) and progression-free survival (PFS) on ovarian cancer patients. We also used Kaplan-Meier to analyze the relationship between miRs and OS in OncoLnc dataset. A total of 15 records were included into the meta-analysis. The expression level of miR-200 family showed significant association with OS (HR = 0.78, 95% CI: 0.64-0.94) and insignificant association with PFS (HR = 0.72, 95% CI: 0.50-1.03). Subgroup analysis revealed that an increased expression level of miR-200c was associated with better OS (HR = 0.59, 95% CI: 0.45-0.74). An increased expression level of miR-200a, miR-200c, and miR-141 was associated with better PFS (miR-200a, HR = 0.59, 95% CI: 0.42-0.75; miR-200c, HR = 0.50, 95% CI: 0.14-0.87, miR-141, HR = 0.38, 95% CI: 0.12-0.63). Similarly, higher expression of miR-30 family was associated with elevated OS/PFS for ovarian cancer (OS, HR = 0.43, 95% CI: 0.13-0.74; PFS, HR = 0.76, 95% CI: 0.64-0.87). The OncoLnc dataset presented that elevated expression level of miR-30d-5p was associated with better OS (n = 470, P = .0197). The meta-analysis reveals that miR-200 family and miR-30 family could be promising prognostic biomarkers of ovarian cancer.

Expression of miR-200a and chemotherapeutic treatment efficacy of glioma.

The correlation between miR-200a expression and chemotherapeutic treatment efficacy of glioma was investigated. There were 45 patients with glioma in observation group whose cancer tissues, paracancerous tissues and serum samples were harvested. Additionally, there were 23 healthy subjects in the control group whose serum samples were also collected. The expression levels of miR-200a in cancer tissues, paracancerous tissues and serum samples were measured by real-time fluorescence-based quantitative polymerase chain reaction (qRT-PCR). The t-test and one-way ANOVA were used to compare the serum levels of miR-200a in patients with different clinical features involving age, sex, tumor location, pathological grade and tumor size. All patients in the observation group received temozolomide-based chemotherapy. The serum levels of miR-200a before chemotherapy were compared between patients who were responsive to chemotherapy [complete response (CR) and partial response (PR)] and patients who were not responsive to chemotherapy [stable disease (SD) and progressive disease (PD)]. The expression level of miR-200a in cancer tissue was significantly lower than that in paracancerous tissue (P<0.05). The serum level of miR-200a in patients in the observation group was lower than that in healthy subjects in the control group (P<0.05). No correlations were found between miR-200a expression and patient age, sex and tumor location (P>0.05), but miR-200a expression was found to correlate with pathological grade and tumor size (P<0.05). The expression levels of miR-200a in serum and cancer tissue in chemotherapy-non-responsive patients (SD and PD) were lower than those in chemotherapy-responsive patients (CR and PR, P<0.05). The serum levels of miR-200a in chemotherapy-responsive patients were lower than those in healthy subjects in the control group (P<0.05). Downregulation of miR-200a was associated with onset and progression of glioma, and changes of miR-200a expression levels in patients were correlated with chemotherapeutic treatment efficacy.

MicroRNA-200a regulates adipocyte differentiation in the domestic yak Bos grunniens

The domestic yak (Bos grunniens) is a culturally important animal that lives at high altitude and is farmed by Tibetan herders for its meat, milk, and other animal by-products. Within the animal, adipose tissue is an important store and source of energy and is used to maintain adequate body temperature during the extended cold seasons. Exploring the biomolecular role of microRNAs (miRNAs) in the regulation of growth, development, and metabolism of yak adipocytes may provide valuable insights into the physiology of adipogenesis in the yak. This study investigated whether and how miR-200a (a miRNA recently reported to promote adipogenesis in ST2 bone marrow stromal cells) regulates adipocyte differentiation in the yak. Expression levels of miR-200a gradually increased during day 0 to day 8 of adipocyte differentiation, and transfection of adipocytes with miR-200a enhanced lipid accumulation and triglyceride content compared to control (un-transfected) adipocytes. We additionally verified (using qRT-PCR analysis) that miR-200a increased the expression of adipocyte-specific genes involved in lipogenic transcription (PPARγ, ELVOL, and C/EBPα), fatty acid synthesis (ACC, ACS, SCD, and FAS), and fatty acid transport (DGAT, LPL, and FABP4). We also found that transfection of adipocytes with miR-200a resulted in suppression of the levels of noncanonical Wnt signaling transcription factors (Wnt5a, TAK1, and NLK). These results indicate that miRNA-200a plays an important role in promoting yak adipocyte differentiation that may operate via the suppression of noncanonical Wnt signaling.

The role of miRNA-200a in the early stage of the mandibular development

BackgroundMicroRNAs are non-coding small RNA molecules that regulate various cellular functions. In this study, we analyzed the function of microRNA-200a (miR-200a) during the development of primary and secondary cartilages in the mandible during the embryonic stage. Materials and methodsMandibular processes were excised from ICR mouse embryos. Meckel’s cartilage and the complex of condylar and angular cartilages were examined as primary and secondary cartilages, respectively. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization were performed to examine the expression of miR-200a during normal mandibular development. Transfection of a miR-200a-mimic and -inhibitor were performed to analyze the effect of miR-200a on mandibular cartilage formation. Prior to transfection, DiI labeling was performed to detect the transfection sites and examine the effect of miR-200a on Meckel’s and condylar cartilage formation. Nine days after transfection, Alcian blue staining and quantification were performed to analyze the formation of the cartilage. ResultsThe expression levels of miR-200a gradually increased from embryonic day 9–14 in mandibular processes. However, the expression levels of miR-200a in Meckel’s cartilage were nearly identical from embryonic day 12–16. The positive hybridization signals were observed in Meckel’s cartilage and mesenchymal condensation of mandibular condylar cartilage. Alcian blue analysis showed a significant decrease in the complex of condylar and angular cartilage formation in the lateral-anterior-miR-200a-mimic transfected samples compared to miR-200a-inhibitor-transfected and control samples. ConclusionmiR-200a negatively regulates the formation of condylar cartilage in early mandibular development.

MicroRNA-200a inhibits cell growth and metastasis by targeting Foxa2 in hepatocellular carcinoma

Background: MicroRNAs (miRNAs) are a class of endogenous, small non-coding RNAs which function as essential posttranscriptional modulators of gene expression tightly involved in a wide range of diseases, including the hepatocellular carcinoma (HCC). Here, the present study was designed to investigate the expression levels and cellular roles of miR-200a in HCC.Methods: Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-200a in serums and cell lines. Bioinformation analysis, the luciferase reporter assay, qRT-PCR and western blotting were employed to validate Foxa2 as a direct target gene of miR-200a. Cell proliferation, migration and invasion were assessed to identify whether miR-200a could regulate the biological behaviors of HCC cells by targeting Foxa2.Results: In this study, a low level of miR-200a was observed in patients' serums and HCC cell lines. Overexpression of miR-200a in HCC cell lines reduced cell proliferation, migration and invasion. In addition, transcription factor forkhead box A2 (Foxa2) was identified as a novel target of miR-200a and downregulated at mRNA and protein levels in miR-200a overexpressed cells. Meanwhile, restoration of Foxa2 significantly reversed the tumor suppressive effects of miR-200a.Conclusions: These findings indicate that miR-200a regulates the proliferation, migration and invasion of HCC cells by targeting Foxa2, suggesting that miR-200a may function as a potential therapeutic molecular for the diagnosis and treatment of the liver cancer.

Expression and Clinical Pathological Significance of miR-200a in Concurrent Cholangiocarcinoma Associated with Hepatolithiasis.

BackgroundApproximately 2–10% of the patients with hepatolithiasis may develop cholangiocarcinoma (CCA). Despite recent advances in the treatment of cancers, the 5-year survival rate for CCA patients currently remains poor, primarily due to early local invasion and distant metastasis of the cancer. This study aimed to investigate miR-200a expression in combined hepatolithiasis and CCA as well as its correlation with the clinical features of CCA.Material/MethodsmiR-200a expression in combined hepatolithiasis and CCA was detected by real-time reverse transcription PCR (qRT-PCR). Its correlation with the clinicopathology of CCA was analyzed by t-tests. The effect of miR-200a on the proliferation CCA cells was determined by MTT assay. The effect of miR-200a on the invasive ability of CCA cells was assessed by Boyden chamber test.ResultsThe expression level of MiR-200a in patients with combined hepatolithiasis and CCA was significantly decreased compared with patients with only hepatolithiasis (P<0.01). Furthermore, miR-200a expression in hepatic duct cancer RBE cells was substantially reduced compared with hepatolithiasis group (P<0.01). Correlation analysis showed that abnormal expression of miR-200a was only associated with the differentiation degree and metastasis of CCA. MiR-200a transfection significantly inhibited the proliferation and invasion of REB cells (P<0.01).ConclusionsMiR-200a may suppress the proliferative and invasive ability of REB cells. The reduced miR-200a expression might be correlated with the development and progression of CCA.

Decreased expression of MicroRNA-200 family in human breast cancer is associated with lymph node metastasis.

MicroRNA-200 family (miR-200f) has been consistently reported to be deregulated and modulate the metastatic process in multiple cancers. In the present study, we detected the expression of miR-200f in breast cancer (BC) tissue and explored its relationships with clinicopathological characteristics, especially with lymph node metastasis. Expression levels of miR-200a, miR-200b, miR-200c, miR-141, and miR-429 in 99 pairs of BC tissues and adjacent normal tissues were measured by real-time quantitative PCR. The correlation between miR-200f level and multiple clinicopathological factors was then examined by Mann-Whitney test, ANOVA, and operating characteristic (ROC) analysis. All members of the miR-200f were down-regulated in BC tissue compared with that in normal adjacent tissue; miR-200a, miR-200b, and miR-200c were highly decreased (p < 0.05), while the differences of miR-141 and miR-429 between patients and the control group were not statistically significant. Furthermore, all five members were found to be distinctly decreased with the incidence of lymph node metastasis (p < 0.05); When the patients were divided into three groups according to the number of lymph node metastasis (0; 1-3; ≥4), a gradual decrease of miR-200f expression was observed with the increasing number of lymph node metastasis; ROC revealed that miR-200b can differentiate patients with lymph node metastasis from those without lymph node metastasis. These observations imply that the down-regulation of miR-200f in human BC is associated with an invasive phenotype, and miR-200b may be useful to estimate the likelihood of the presence of pathologically positive lymph nodes.

Expression of serum miR-200a, miR-200b, and miR-200c as candidate biomarkers in epithelial ovarian cancer and their association with clinicopathological features.

MicroRNAs (miRs) have been implicated in the etiology of various human cancers. The aim of this study was to investigate the association of the expression of three members--miR 200a, miR 200b, and miR 200c belonging to the miR-200 family with clinicopathological characteristics and their impact on the progression of epithelial ovarian cancer (EOC). Total RNA from serum was isolated by Trizol method, polyadenylated, and reverse transcribed into cDNA. Expression levels of miR-200a, miR-200b, and miR-200c were detected by using miRNA qRT-PCR. We measured miR expression in 70 serum samples of EOC patients with matched controls using U6 snRNA as a reference. Levels of miR expression was compared with distinct clinicopathological features. Expression of miR-200a was found to be greater than six-fold (p = 0.01), miR-200b and miR-200c greater than three-fold (p = 0.01) in comparison with matched normal controls. Association of miRNA expression with clinicopathological factors and progression was statistically evaluated. The expression levels of miR-200a and miR-200c were found to be significantly associated with disease progression (p = 0.04 and p < 0.001, respectively). miR-200a overexpression was found be associated with tumor histology and stage. Patients with lymph node metastasis showed significant elevation of miR-200c (p = 0.006). The AUC in ROC curve also indicated that serum levels of miR-200a and miR-200c might be worthwhile as a diagnostic tool in the near future. Our findings suggest that miR-200a, miR-200b, and miR-200c overexpressions are associated with the aggressive tumor progression and be recognized as reliable markers to predict the prognosis and survival in EOC patients.