Roles of ghrelin, hepcidin and HIF-2α in iron metabolism in iron deficiency anemia.

OBJECTIVES: This study investigates the roles of HIF-2α, hepcidin, and ghrelin in iron deficiency anemia (IDA), the most widespread nutritional disorder globally. MATERIAL AND METHODS: Fifty IDA patients (18-50 years, BMI 19-25) and 40 healthy volunteers were studied. Hemoglobin, ferritin, hepcidin, HIF-2α, and ghrelin levels were analyzed. RESULTS: IDA patients showed lower hemoglobin, ferritin, hepcidin, and ghrelin levels than the control group, but HIF-2α levels were similar. Positive correlations were observed in both groups between hepcidin and HIF-2α (p < 0.001), hepcidin and ghrelin (p < 0.001), and HIF-2α and ghrelin (p < 0.001). Hemoglobin was correlated positively with HIF-2α, and ferritin was correlated positively with HIF-2α in the patient group. CONCLUSION: The study suggests that the low hepcidin levels in IDA patients enhance iron absorption. The lack of significant HIF-2α level differences may be due to the absence of chronic hypoxia in current hemoglobin levels of IDA patients. Moreover, the low ghrelin levels in patients and the correlations between ghrelin, hepcidin, and HIF-2α in both groups indicate their involvement in iron metabolism.

Role of mir-33a, mir-203b, mir361-3p, and mir-424 in hepatocellular carcinoma

Background/aim: Hepatocellular carcinoma (HCC) is one of the most aggressive cancer types. MicroRNAs (miRNAs) are small noncoding regulatory RNAs that function posttranscriptionally. miRNA deregulation was observed in the development and progression of HCC. In this study, we aimed to investigate the expression levels of four miRNAs (mir-33a, mir-203b, mir361-3p, and mir-424) in HCC patients in comparison to healthy individuals. Materials and methods: Venous blood samples were collected from both HCC patients and healthy individuals. In order to determine the relative expression levels of mir-33a, mir-203b, mir361-3p, and hsa-mir-424 in HCC patients, probe-based quantitative real time PCR (qRT-PCR) was performed. The cycle threshold (Ct) results were analyzed according to the 2−∆∆Ct method and statistical analyses were performed by SPSS Statistics version 15 for Windows. Results: qRT-PCR analysis revealed that the expression levels of mir-33a (fold change: 7.3 and P < 0.001), mir-203b (fold change: 4.6 and P < 0.001), and mir361-3p (fold change: 5.1 and P < 0.001)were downregulated compared to healthy individuals and mir-424 did not show any significant change between HCC patients and controls. Conclusion: Our results indicated that mir-33a, mir-203b, and mir-361-3p may significantly contribute to tumor pathogenesis in HCC and have potential to be used as a noninvasive biomarker for cancer therapy.

Tumor suppressor and oncogenic role of long non-coding RNAs in cancer.

Non-coding RNAs are RNA molecules that are not translated into the protein, making up the vast majority of the human genome. Long non-coding RNAs (lncRNA) are in the RNA group that has longer than 200 nucleotides, and non-protein coding transcripts. In recent years, the potential has attracted considerable attention as new important biological regulators. LncRNAs play a critical role in regulating the activity and localization of proteins, processing the production of small RNAs, and processing other RNAs. They are also involved in cell differentiation, cell cycle, proliferation, apoptosis, migration and invasion by modulation of gene expression. Abnormal expression of LncRNAs has an important role in the function of oncogenes and tumor suppressor genes. Recently, there has been an increasing number of studies on the tumorigenic effects of specific lncRNAs in the initiation and progression of cancer. In this review, general information about lncRNAs is provided, including the biological importance of lncRNAs in cancer diseases and their potential development in therapeutic applications.

MicroRNAs as prognostic markers in prostate cancer.

BACKGROUND: Prostate cancer (PCa) is the most commonly diagnosed malignancy in men who are especially over the age of 50 years in the western countries. Currently used therapeutic modalities mostly fail to give positive clinical outcomes and nearly 30% of the PCa patients eventually develop clinical recurrence. Therefore, understanding the underlying mechanisms of PCa progression is of paramount importance to help determining the course of disease. In this study, we aimed at profiling the differentially expressed microRNAs in recurrent PCa samples. METHODS: We profiled the microRNA expression of 20 recurrent and 20 non-recurrent PCa patients with microRNA microarray, and validated the differential expression of significantly deregulated microRNAs in 40 recurrent and 39 non-recurrent PCa specimens using quantitative reverse-transcription PCR (qRT-PCR). Data were statistically analyzed using two-sided Student's t-test, Pearson Correlation test, Receiver operating characteristic (ROC) analysis. RESULTS: Our results demonstrated that a total of 682 probes were significantly deregulated in recurrent versus non-recurrent PCa specimen comparison. Among those, we confirmed the significant downregulation of miR-424 and upregulation of miR-572 with further qRT-PCR analysis in a larger sample set. Further ROC analysis showed that these microRNAs have enough power to distinguish recurrent specimens from non-recurrent ones on their own. CONCLUSIONS: Here, we report that differential expression of miR-424 and miR-572 in recurrent PCa specimens can serve as novel biomarkers for prediction of PCa progression.

Unsuspected Diagnosis of Uterine Leiomyosarcoma after Laparoscopic Myomectomy in an Isolated Bag.

Minimally invasive techniques are generally applied for patients suspected of having benign fibroids if medical treatment is insufficient. On the other hand, sometimes some occult carcinomas of uterus like leiomyosarcomas may be reported for the patients' applied morcellation. This condition is rare but outcomes are clinically significant. Fragmentation of occult sarcoma in the abdominal cavity without isolation bag results in widespread and poor survival. In this article, we report a case of 37-year-old woman suffering from pain due to unexpected leiomyosarcoma. Laparoscopic myomectomy was performed with power morcellation in an isolated bag. Although isolation bag is generally reported to be preventive, recurrence of sarcoma was seen at 5th month of follow-up. Even though morcellation within a bag seems to block wide spreading, dispersion of tumor cannot be stopped and more investigations have to be done.

A Meta-Analysis: Identification of Common Mir-145 Target Genes that have Similar Behavior in Different GEO Datasets.

BACKGROUND: MicroRNAs, which are small regulatory RNAs, post-transcriptionally regulate gene expression by binding 3'-UTR of their mRNA targets. Their deregulation has been shown to cause increased proliferation, migration, invasion, and apoptosis. miR-145, an important tumor supressor microRNA, has shown to be downregulated in many cancer types and has crucial roles in tumor initiation, progression, metastasis, invasion, recurrence, and chemo-radioresistance. Our aim is to investigate potential common target genes of miR-145, and to help understanding the underlying molecular pathways of tumor pathogenesis in association with those common target genes. METHODS: Eight published microarray datasets, where targets of mir-145 were investigated in cell lines upon mir-145 over expression, were included into this study for meta-analysis. Inter group variabilities were assessed by box-plot analysis. Microarray datasets were analyzed using GEOquery package in Bioconducter 3.2 with R version 3.2.2 and two-way Hierarchical Clustering was used for gene expression data analysis. RESULTS: Meta-analysis of different GEO datasets showed that UNG, FUCA2, DERA, GMFB, TF, and SNX2 were commonly downregulated genes, whereas MYL9 and TAGLN were found to be commonly upregulated upon mir-145 over expression in prostate, breast, esophageal, bladder cancer, and head and neck squamous cell carcinoma. Biological process, molecular function, and pathway analysis of these potential targets of mir-145 through functional enrichments in PPI network demonstrated that those genes are significantly involved in telomere maintenance, DNA binding and repair mechanisms. CONCLUSION: As a conclusion, our results indicated that mir-145, through targeting its common potential targets, may significantly contribute to tumor pathogenesis in distinct cancer types and might serve as an important target for cancer therapy.

Identification of miR-139-5p as a saliva biomarker for tongue squamous cell carcinoma: a pilot study.

BACKGROUND: Of all human oral carcinomas, 41 % are localized to the tongue. Despite considerable improvements in both diagnosis and treatment, tongue squamous cell carcinoma (TSCC) has remained one of the most lethal types of cancer. Here, we aimed at identifying a salivary microRNA (miRNA) expression signature specific for TSCC patients. METHODS: To identify putative diagnostic biomarkers, we compared the miRNA expression profiles of saliva samples from three TSCC patients and four healthy control individuals using an Agilent miRNA microarray platform (V19). Three of the differentially expressed miRNAs identified were selected for further validation using quantitative reverse-transcription PCR (qRT-PCR) in saliva samples from 25 TSCC patients and 25 healthy control individuals. RESULTS: Through microarray-based expression profiling, we found that 419 miRNAs were deregulated in the saliva samples from the TSCC patients compared to those from the healthy control individuals tested. Subsequent qRT-PCR analysis revealed that the expression level of miR-139-5p was significantly reduced in the TSCC validation samples compared to the controls. Further analysis of post-operative saliva samples derived from TSCC patients revealed that the miR-139-5p expression levels had turned back to normal again. In addition, we found that miR-139-5p exhibited enough power to discriminate pre-operative TSCC patients from both normal individuals (AUC: 0.805) and post-operative TSCC patients (AUC: 0.713), thereby underscoring its diagnostic potential. CONCLUSIONS: From our results we conclude that saliva can be used as a feasible source for routine TSCC diagnostics and that miR-139-5p may serve as a potential biomarker for early TSCC detection.

The role of ATP-binding cassette transporter genes in the progression of prostate cancer.

BACKGROUND: Prostate cancer (PCa) is the most commonly diagnosed neoplasm and the second leading cause of cancer-related death among men in developed countries. There is no clear evidence showing the success of current screening tests in reducing mortality of PCa. In this study, we aimed to profile expressions of nine ABC transporters, ABCA5, ABCB1, ABCB6, ABCC1, ABCC2, ABCC3, ABCC5, ABCC10, and ABCF2, in recurrent, non-recurrent PCa and normal prostate tissues. METHODS: A total of 77 (39 recurrent, 38 non-recurrent) radical prostatectomy and 20 normal prostate samples, obtained from Baylor College of Medicine Prostate Cancer program, were included into the study and divided into two independent groups as test and validation sample sets. Differential expression of selected ABC transporters was assessed using quantitative real-time PCR (qRT-PCR). Pearson's correlation test, receiver operating characteristics (ROC) analysis and Kaplan-Meier test were used for statistical analysis. RESULTS: QRT-PCR results demonstrated the elevated expression of ABCA5, ABCB1, ABCB6, ABCC1, and ABCC2 as well as reduced expression of ABCC3 in PCa samples compared to normal prostate tissues. In addition, we found deregulation of ABCB1, ABCB6, ABCC3, and ABCC10 in recurrent PCa samples and validated differential expression of ABCB6, ABCC3, and ABCC10 in recurrent PCa compared to non-recurrent PCa. Pearson's correlation, ROC and Kaplan-Meier analysis revealed the power of these three ABC transporters for estimating prognosis of PCa. CONCLUSIONS: We demonstrated differential expression of ABC transporters both in tumor versus normal and recurrent versus non-recurrent comparisons. Our data suggest ABCB6, ABCC3, and ABCC10 as valuable predictors of PCa progression.

Overexpression of miR-145-5p inhibits proliferation of prostate cancer cells and reduces SOX2 expression.

We aimed to perform functional analysis of miR-145-5p in prostate cancer (PCa) cells and to identify targets of miR-145-5p for understanding its role in PCa pathogenesis. PC3, DU145, LNCaP PCa, and PNT1a nontumorigenic prostate cell lines were utilized for functional analysis of miR-145-5p. Its overexpression caused inhibition of proliferation through apoptosis and reduced migration in PCa cells. SOX2 expression was significantly decreased in both mRNA and protein level in miR-145-5p-overexpressed PCa cells. We proposed that miR-145-5p, being an important regulator of SOX2, carries a crucial role in PCa tumorigenesis.

MiR-221 as a pre- and postoperative plasma biomarker for larynx cancer patients.

OBJECTIVE: In order to identify a plasma microRNA (miRNA) signature of larynx cancer (LCa), we examined miRNAs profile of plasma samples obtained from 30 LCa patients (preoperative and postoperative serum samples) and 30 healthy controls. STUDY DESIGN: Basic science research study. METHODS: MicroRNA profiling of eight plasma samples (four from preoperative, four from control individuals) were performed using miRNA microarray. Two of the significantly deregulated miRNAs were selected for further confirmation in the remaining samples using quantitative reverse-transcription polymerase chain reaction (qRT-PCR). RESULTS: Microarray profiling and qRT-PCR analysis showed that miR-221 was upregulated in LCa plasma samples. Further qRT-PCR analysis demonstrated that miR-221 was at normal levels in postoperative plasma samples. CONCLUSIONS: Plasma miR-221 may have a potential as a novel diagnostic/prognostic marker and might be considered as a therapeutic target in LCa. LEVEL OF EVIDENCE: N/A.

Identification of microRNAs differentially expressed in prostatic secretions of patients with prostate cancer.

Prostate cancer (PCa) is one of the leading causes of cancer deaths in men. Since there are limited treatment options available for the advanced tumors, there is an urgent need for novel diagnostic tools for PCa. Prostate secretion samples (PSS) from 23 PCa and 25 benign prostate hyperplasia (BPH) patients were obtained from Urology Department of Bagcilar Educational and Research Hospital (Istanbul). MicroRNA (miRNA) profiling of eight PSS (four from BPH, four from PCa patients) was performed using microarray. Four of significantly deregulated miRNAs were further confirmed using quantitative reverse-transcription PCR (qRT-PCR). Statistical analysis was performed using Student's t-test. ROC curves were plotted with SPSS-15.0. In this study, we aimed to identify a miRNA expression signature that could be used to distinguish PCa from BPH. MiRNA profiling of four PCa and four BPH patients with microarray revealed that miR-361-3p, miR-133b and miR-221 were significantly downregulated and miR-203 was upregulated in PSS of PCa patients. Further qRT-PCR analysis confirmed the altered expressions of these four miRNAs in PSS of 23 PCa and 25 BPH patients. Four miRNAs, together and individually have much power (AUC; 0.950) than PSA has (AUC; 0.463) to discriminate PCa from BPH patients. We have shown for the first time in the literature the presence of miRNAs in the PSS. We suggest PSS as a powerful non-invasive source for evaluation of prognosis in PCa, since prostate massages can be easily applied during routine examination. Our results showed that certain differentially expressed miRNAs in PSS could be used as diagnostics markers.

Differential expression of stem cell markers and ABCG2 in recurrent prostate cancer.

BACKGROUND: Prostate cancer (PCa) is the second most common tumor type related to mortality in males in the developed countries. Studies have demonstrated that therapeutic tools mostly ineffective to give positive outcome especially for PCa. Cancer stem cells are composed of a small cell population, which are supposed to have roles in tumorigenesis, metastasis, and tumor recurrence after chemo-radiotherapy. The aim of this research is to investigate expressions of stem cell markers in recurrent PCa and non-recurrent PCa tumors as well as in adjacent normal prostate tissues. METHODS: We compared the expression of important stemness regulators like SOX2, OCT4, KLF4, and ABCG2 in recurrent, non-recurrent PCa and adjacent normal tissue samples using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: Our results demonstrated that SOX2 and OCT4 are strongly overexpressed in PCa samples. Recurrent PCa samples are markedly positive for stem cell markers SOX2, OCT4, and KLF4. Furthermore, non-recurrent PCa samples presented low levels of ABCG2, a multidrug resistance protein, compared to both normal and recurrent samples, which might be associated with chemo-sensitivity. CONCLUSIONS: Enhanced expression of ABCG2 and stem cell markers including SOX2, OCT4, and KLF4 in the recurrent PCa tissues postulates the suggestion that enrichment for cells with stem cell characteristics in these tissues might be playing a critical role for chemoresistance and recurrence of cancer.

miR-1 and miR-133b are differentially expressed in patients with recurrent prostate cancer.

Prostate cancer (PCa) is currently the most frequently diagnosed malignancy in the western countries. It is more prevalent in older men with 75% of the incident cases above 65 years old. After radical prostatectomy, approximately 30% of men develop clinical recurrence with elevated serum prostate-specific antigen levels. Therefore, it is important to unravel the molecular mechanisms underlying PCa progression to develop novel diagnostic/therapeutic approaches. In this study, it is aimed to compare the microRNA (miRNA) profile of recurrent and non-recurrent prostate tumor tissues to explore the possible involvement of miRNAs in PCa progression. Total RNA from 41 recurrent and 41 non-recurrent PCa tissue samples were used to investigate the miRNA signature in PCa specimens. First of all, 20 recurrent and 20 non-recurrent PCa samples were profiled using miRNA microarray chips. Of the differentially expressed miRNAs, miR-1, miR-133b and miR-145* were selected for further validation with qRT-PCR in a different set of 21 recurrent and 21 non-recurrent PCa samples. Data were statistically analyzed using two-sided Student's t-test, Pearson Correlation test, Receiver operating characteristic analysis. Our results demonstrated that miR-1 and mir-133b have been significantly downregulated in recurrent PCa specimens in comparison to non-recurrent PCa samples and have sufficient power to distinguish recurrent specimens from non-recurrent ones on their own. Here, we report that the relative expression of miR-1 and mir-133b have been significantly reduced in recurrent PCa specimens in comparison to non-recurrent PCa samples, which can serve as novel biomarkers for prediction of PCa progression.

Experience of tubo-ovarian abscess in western Turkey.

OBJECTIVE: To investigate the clinical and laboratory parameters, treatments, and complications of patient with tubo-ovarian abscess (TOA). METHODS: Data for 296 patients diagnosed with TOA (clinically and sonographically) between January 2005 and December 2012 were retrospectively reviewed at 3 tertiary referral hospitals in Turkey. Patients were compared on the basis of TOA size, demographic characteristics, clinical and sonographic presentation, and laboratory findings. RESULTS: Seventy-six patients (25.7%) underwent surgery because antibiotic treatment was unsuccessful. The mean abscess size was larger and the mean C-reactive protein (CRP) level and the erythrocyte sedimentation rate (ESR) were higher among patients who required surgery. The ESR had a diagnostic value of 83.6%, and a specificity and sensitivity of 73.7% and 82.7%, respectively, for the need for surgical intervention, based on a cut-off value of 63.0mm/hour. The CRP level had a diagnostic value of 80.4%, a specificity of 82.3%, and a sensitivity of 65.8% based on a cut-off value of 21.0mg/L. CONCLUSION: The combined use of the sonographic TOA diameter and laboratory parameters (ESR and CRP level) can aid clinical treatment decisions and improve the prediction of the outcome of medical TOA treatment.

MicroRNA profiling in lymphocytes and serum of tyrosinemia type-I patients.

Tyrosinemia type-I results from lack of fumarylacetoacetate hydrolase (FAH), which is a liver enzyme and also shown to be present in lymphocytes, fibroblasts, and cultured amniotic fluid cells. In young infants, symptoms of untreated Tyrosinemia type-I are restricted to severe liver involvement. Later in the first year; however, it is known to be present with liver and renal tubular dysfunction associated with growth failure and rickets. MicroRNAs are small regulatory RNAs that function post-transcriptionally. They target commonly 3'-UTR of the mRNAs and inhibit protein expression by either blocking the synthesis or causing degradation of the mRNAs. MiRNA deregulation was observed in a variety of pathologic conditions but their roles in metabolic diseases were remained unsolved. We studied 6 patients with classical phenotypes of Tyrosinemia type-I. To identify possible miRNAs targeting FAH transcripts, microarray profiling of 961 miRNAs for lymphocytes and serum is performed. Computational algorithms are used for prediction of putative mRNA-miRNA interactions. A number of deregulated miRNAs, targeting the non-conserved sites on FAH transcripts were found. Besides, there are some miRNAs that are similarly altered both in lymphocytes and serum, possibly contributing to the disease phenotype. Since miRNAs may have an active role in the enzymatic pathway of tyrosine catabolism, characterizing miRNA profile in fibroblasts of tyrosinemia patients is also important because miRNAs would have distinctive role in disease pathogenesis and they are promising for future therapeutic studies.

The healing-promoting effect of saliva on skin burn is mediated by epidermal growth factor (EGF): role of the neutrophils.

Local skin trauma induces inflammatory responses resulting in local tissue and distant organ injury. EGF, a polypeptide hormone, mainly produced in saliva, is one of the major accelerators in wound healing. Wistar albino rats of both sexes received either bovine serum albumin or EGF (10 microg/kg) subcutaneously before a circular (18 mm diameter) partial thickness burn was induced. Afterwards, some rats were placed in separate cages to prevent licking, while the others were caged together to allow wound-licking. Treatments were continued for 5 more days and on the 5th day animals were decapitated. Histopathological analysis of skin damage and dermal myeloperoxidase (MPO) activity, as an index for neutrophil activity, were evaluated. Oxidant injury to the liver and intestines was determined by measuring glutathione (GSH) and malondialdehyde (MDA) levels, as well as MPO activity. The results demonstrate that healing of the burn wound on the skin is accelerated by both wound-licking and EGF administration, which also attenuated tissue neutrophil accumulation, suggesting the role of neutrophils as the source of mediators involved in delayed epithelial regeneration. Moreover, local dermal burn results in oxidant injury to the liver, concomitant with significant elevations in hepatic and intestinal GSH levels. Exogenous administration of EGF at physiological doses had no effect on inflammatory responses of the distant organs, while allowing the rats to lick the wound reduced the oxidant injury to the liver. Since saliva or EGF enhances skin wound healing, topical use of EGF-rich artificial saliva merits consideration for its use in burn patients.