Evaluation of vascular endothelial growth factor A and leukemia inhibitory factor expressions at the time of implantation in diabetic rats following treatment with Metformin and Pioglitazone

Background: Implantation requires intimate crosstalk between the embryo and uterus for a successful establishment of pregnancy. Type 2 diabetes mellitus may lead to implantation failure. The effect of diabetes and its therapeutic drugs on implantation is still largely unclear. Objective: To assess the endometrial expression changes of vascular endothelial growth factor A (VEGFA) and leukemia inhibitory factor (LIF), at the time of implantation in diabetic rats following treatment with Metformin and Pioglitazone. Materials and Methods: Twenty-eight 6-8-wk-old Wistar female rats weighing 200- 250 gr were divided into four groups (n = 7/each). Type 2 diabetes was induced and Metformin and Pioglitazone were applied for 4 wk. The expression of VEGFA and LIF was measured by real-time reverse transcription-polymerase chain reaction and Western blot. Results: The relative expression of VEGFA transcript was higher in the diabetic (p = 0.02) and Metformin-treated (p = 0.04) rats compared to the control group. Furthermore, the VEGFA transcript level significantly reduced in Pioglitazone-treated diabetic rats (p = 0.03). LIF expression was elevated in the Metformin- and the Pioglitazone-treated rats and reduced in the diabetic group in comparison with the control group. Compared to the diabetic rats, the expression of LIF was significantly elevated in the Metformin- (p = 0.01) and Pioglitazone-treated (p = 0.03) groups. Conclusion: The expressions of LIF and VEGFA were altered in diabetic rats during implantation which may be associated with diabetic-related infertility. Pioglitazone is able to restore the VEGFA and LIF expressions to their baseline levels more efficiently than Metformin. Key words: Embryo implantation, Leukemia inhibitory factor, Vascular endothelial growth factor A, Metformin, Pioglitazone, Rats

Down-regulation of circular RNA ITCH and circHIPK3 in gastric cancer tissues

Background/aim: Gastric cancer (GC) is one of the major causes of cancer mortality worldwide. As a novel type of endogenous noncoding RNAs, circular RNAs (circRNAs) are formed by a covalent link between 5’ and 3’ ends. They are very stable and abundant in eukaryotes. As there were no reported studies on the expression profiles of circular RNA ITCH (cir-ITCH) and circHIPK3 in GC, in the current study, we aimed to delineate the expression profiles and clinicopathological relevance of these two circRNAs in GC tissues compared to their paired adjacent noncancerous tissues. Materials and methods: Quantitative real-time polymerase chain reaction was performed to evaluate cir_ITCH and circHIPK3 expression in 30 paired gastric cancer tissues. The clinicopathological relevance of these two circular RNAs’ expression levels with gastric cancer was further examined. Results: Our results showed that the expression of cir_ITCH and circHIPK3 were significantly downregulated in GC tumoral tissues compared with their paired adjacent nonneoplastic counterparts. Further analyses showed that cir_ITCH and circHIPK3 expression levels were related with numerous clinicopathological features of tumoral tissues. Conclusion: Cir_ITCH and circHIPK3 may have imperative roles in GC and serve in the future as potential prognostic biomarkers in GC. Key words: Gastric cancer, circular RNAs, cir_ITCH, circHIPK3, gene expressio

Evaluation of the expression of Hottip long noncoding RNA in the B16F10 murine melanoma cell line

زمینه و هدف: ملانوما یکی از انواع سرطان پوست است که نسبت به سایر سرطان های پوست خطرناک تر می باشد. یکی از عوامل موثر در ایجاد سرطان ها، گروهی از RNA های غیر کد کننده اند که به نام lncRNA (long noncoding RNA) شناخته می شوند. این مولکول های غیر کد کننده بیش از 200 باز طول دارند و به عنوان یک تنظیم کننده در پیشرفت سرطان عمل می کنند Hottip (HOXA transcript at the distal tip) یک lncRNA ی بین ژنی است که از انتهای '5 لوکوس Hoxa رونویسی می شود و ژن های انتهای '5 این لوکوس را فعال می کند. هدف از این مطالعه، بررسی میزان بیان این مولکول RNA در رده ی سلولی B16F10 ملانومای موشی بود. روش بررسی: در این مطالعه بیان ژن Hottip با انجام تکنیک RT-PCR به صورت کیفی روی رده ی سلولی B16F10 ملانومای موشی بررسی شد. بدین منظور، از رده ی سلولی، RNA کل استخراج و سنتز cDNA انجام گرفت. با استفاده از پرایمرهای اختصاصی طراحی شده، ژن های Hottip و β2m تکثیر گردیدند. یافته ها: نتیجه مطالعه حاضر نشان دهنده ی عدم بیان ژن Hottip موشی در رده ی سلولی B16F10 ملانومای موشی است. نتیجه گیری: در حالی که بر اساس مطالعات صورت گرفته در سرطان های انسانی انتظار می رفت Hottip افزایش بیان داشته باشد، Hottip موشی در رده ی سلولی B16F10 ملانومای موشی بیانی نشان نداد

Heavy metals (Hg,Cd,Pb,Ni,Cu) concentrations in Euryglossa orientalis and sediments from Khur-e-Musa Creek in Khuzestan Province

Heavy metals contamination (Hg,Cd,Pb,Ni,Cu) in muscle of the fish Euryglossa orientalis and in sediments was assessed in 2007 in Khur-e-Musa Creek (Ahmadi and Ghanam). In total, 30 fish specimens and 18 sediment samples were collected and analyzed. Flame Atomic Absorption Spectrophotometer was used to determine contamination of the specimens with Cd, Pb, Ni, Cu, and cold vapor method was applied for Hg. Results showed 2.35, 0.99, 1.32, 14.48 and 5.71µg/g dry weight of the fish for Hg, Cd, Pb, Ni and Cu in muscle tissue, respectively. Metal levels in the muscle tissue were compared with standard values such as those of the World Health Organization (WHO), British Ministry of Agriculture, Fisheries and Food (MAFF), Australia National Health and Medical Research Council (NHMRC) and American Food and Drug Administration (FDA), based on which only Hg, Cd, and Ni showed higher than standard levels in Khur-e-Musa Creek (Ahmadi and Ghanam). Results showed 4.76, 2.52, 18.64, 119.91, 31.23µg/g dry weight for Hg, Cd, Pb, Ni and Cu in sediments, respectively

MT1XT20 single quasi-monomorphic mononucleotide marker for detection of microsatellite instability in iranian patients with hereditary nonpolyposis colorectal cancer (HNPCC)

Background: Colorectal malignancies with high microsatellite instability (MSI-H), either hereditary or sporadic, demonstrate better prognosis, altered response to fluorouracil (5FU) chemotherapy and altered operative approach. It is now recommended to perform MSI testing for all new cases of colorectal cancers regardless of being categorized as hereditary or sporadic. This study aimed to evaluate MT1XT20 mononucleotide marker in Iranian patients with hereditary nonpolyposis colorectal cancer (HNPCC). The samples were further characterized using Promega five-marker MSI testing panel and immunohistochemical (IHC) technique. Methods: MT1XT20 mononucleotide marker and commercially available kit (Promega, USA) incorporating five quasi-monomorphic markers were studied in 20 cases of HNPCC using polymerase chain reaction (PCR) technique. IHC was performed to evaluate the status of all four important mismatch repair (MMR) proteins, too. Findings: Eight (40%), seven (35%) and five (25%) cases showed MSI using Promega kit, IHC and MT1XT20, respectively. Among the markers included in Promega kit, BAT26 marker with instability in all 8 samples (100%) was the most instable marker. NR24 and NR21 markers showed instability in 7 cases (87.5%); BAT25 and MONO 27 markers were instable in 6 (75.0%) and 5 (62.5%) specimens, respectively. Conclusion: Although MT1XT20 is considered as a valid single marker in Italian population, it seems this is not hold true about the Iranian patients. Instead, BAT26 among the markers included in Promega MSI testing was shown instability in all 8 samples of MSI-H colorectal cancer (CRC). Therefore, it may be concluded that BAT26 alone is as efficient as the cohort of five markers in Iranian patients. © 2016, Isfahan University of Medical Sciences(IUMS). All rights reserved

Evaluation of MT1XT20 single quasi-monomorphic mononucleotide marker for characterizing microsatellite instability in persian lynch syndrome patients

Background: Colorectal malignancies with high microsatellite instability (MSI-H), either hereditary (Lynch syndrome) or sporadic, demonstrate better prognosis and altered response to 5FU chemotherapy. It is now recommended to perform MSI testing for all new cases of colorectal cancer regardless of being categorized as hereditary or sporadic. For MSI detection, immunohistochemistry or PCR-based protocols using a cohort of various sets of STR markers are recommended. Here we aimed to evaluate a simplified protocol using just a single STR marker, MT1XT20 mononucleotide repeat, for detection of MSI in Lynch syndrome patients. A Promega five-marker MSI testing panel and immunohistochemistry (IHC) were used as the gold standard in conjunction with MT1XT20. Materials and Methods: Colorectal patients with a positive history of familial cancers were selected by evaluating medical records. Based on Amsterdam II criteria for Lynch syndrome 20 families were short listed. DNA was extracted from formalin fixed paraffin embedded tumour and adjacent normal tissues resected from the index case in each family. Extracted DNA was subjected to MT1XT20 mononucleotide marker analysis and assessment with a commercially available five marker MSI testing kit (Promega, USA). IHC also was performed on tissue sections and the results were compared with PCR based data. Results: Eight (40%), seven (35%) and five (25%) cases were MSI positive using with the Promega kit, IHC and MT1XT20, respectively. Among the markers included in Promega kit, BAT26 marker showed instability in all 8 samples. NR24 and NR21 markers showed instability in 7 (87.5%), and BAT25 and MONO 27 in 6 (75%) and 5 (62.5%). Conclusions: Although MT1XT20 was earlier reported as a valid standalone marker for MSI testing in CRC patients, we could not verify this in our Iranian patients. Instead BAT26 among the markers included in Promega MSI testing kit showed instability in all 8 MSI-H CRC samples. Therefore, it seems BAT26 could act well as a single marker for MSI testing in Iranian CRC patients

Diabetes mellitus increased integrins gene expression in rat endometrium at the time of embryo implantation

Background: Diabetes mellitus deeply changes the genes expression of integrin (Itg) subunits in several cells and tissues such as monocytes, arterial endothelium, kidney glomerular cells, retina. Furthermore, hyperglycemia could impress and reduce the rate of successful assisted as well as non-assisted pregnancy. Endometrium undergoes thorough changes in normal menstrual cycle and the question is: What happens in the endometrium under diabetic condition? Objective: The aim of the current study was to investigate the endometrial gene expression of α3, α4, αv, Itg β1 and β3 subunits in diabetic rat models at the time of embryo implantation. Materials and Methods: Twenty-eight rats were randomly divided into 4 groups: control group, diabetic group, pioglitazone-treated group, and metformin-treated group. Real-time PCR was performed to determine changes in the expression of Itg α3, α4, αv, β1, and β3 genes in rat’s endometrium. Results: The expression of all Itg subunits increased significantly in diabetic rats’ endometrium compared with control group. Treatment with pioglitazone significantly reduced the level of Itg subunits gene expression compared with diabetic rats. While metformin had a different effect on α3 and α4 and elevated these two subunits gene expression. Conclusion: Diabetes mellitus significantly increased the expression of studied Itg subunits, therefore untreated diabetes could be potentially assumed as one of the preliminary elements in embryo implantation failure

Clinical Trial: CYP2D6 Related Dose Escalation of Tamoxifen in Breast Cancer Patients With Iranian Ethnic Background Resulted in Increased Concentrations of Tamoxifen and Its Metabolites

Introduction: The polymorphic enzyme cytochrome P450 2D6 (CYP2D6) catalyzes a major step in the bioactivation of tamoxifen. Genotyping of clinically relevant CYP2D6 alleles and subsequent dose adjustment is a promising approach to individualize breast cancer therapy. The aim of this study was to investigate the relationship between the plasma levels of tamoxifen and its metabolites and different CYP2D6 genotypes under standard (20 mg/day) and dose-adjusted therapy (Registration ID in Iranian Registry of Clinical Trials: IRCT2015082323734N1).Materials and Methods: Using TaqMan® assays common alleles of CYP2D6 (∗1, ∗2, ∗4, ∗5, ∗6, ∗10, ∗17, and ∗41) and gene duplication were identified in 134 breast cancer patients. Based on CYP2D6 genotypes patients with an activity score 1 (n = 15) and 0–0.5 (n = 2) were treated with tamoxifen adjusted dosage of 30 and 40 mg/day, respectively. The concentration of tamoxifen and its metabolites before and after 4 and 8 months of dose adjustment were measured using LC-MS/MS technology.Results: At baseline, (Z)-endoxifen plasma concentrations (33 ± 15.5, 28.1 ± 14, 26.6 ± 23.4, 14.3 ± 8.6, and 10.7 ± 5.5 nmol/l for EM/EM, EM/IM, EM/PM, IM/IM and PM/PM, respectively) and the metabolic ratio (Z)-Endoxifen/N-desmethyltamoxifen (0.0558 ± 0.02, 0.0396 ± 0.0111, 0.0332 ± 0.0222, 0.0149 ± 0.0026, and 0.0169 ± 0.0177 for EM/EM, EM/IM, EM/PM, IM/IM, and PM/PM, respectively) correlated with CYP2D6 genotype (Kruskal–Wallis p = 0.013 and p < 0.0001, respectively). Dose escalation to 30 and 40 mg/day in patients with a CYP2D6 activity score of 1 (n = 15) and 0–0.5 (n = 2) resulted in a significant increase in (Z)-endoxifen plasma levels (22.17 ± 24.42, 34.43 ± 26.54, and 35.77 ± 28.89 nmol/l at baseline, after 4 and 8 months, respectively, Friedman p = 0.0388) along with the plasma concentrations of tamoxifen and its other metabolites. No severe side effects were recorded during dose escalation.Conclusion: For the first time, we show the feasibility of dose escalation of tamoxifen in breast cancer patients with compromised CYP2D6 activity and Iranian ethnic background to increase the plasma concentrations of (Z)-endoxifen

MicroRNAs-mediated regulation of the differentiation of dental pulp-derived mesenchymal stem cells: a systematic review and bioinformatic analysis

Background: Human dental pulp-derived mesenchymal stem cells (hDP-MSCs), which include human dental pulp stem cells (hDPSCs) and stem cells from human exfoliated deciduous teeth (SHEDs), are promising cell sources for regenerative therapies. Nevertheless, a lack of knowledge relating to the mechanisms regulating their differentiation has limited their clinical application. microRNAs (miRNAs) are important regulatory molecules in cellular processes including cell differentiation. This systematic review aims to provide a panel of miRNAs that regulate the differentiation of hDP-MSCs including hDPSCs and SHEDs. Additionally, bioinformatic analyses were conducted to discover target genes, signaling pathways and gene ontologies associated with the identified miRNAs. Methods: A literature search was performed in MEDLINE (via PubMed), Web of Science, Scopus, Embase and Cochrane Library. Experimental studies assessing the promotive/suppressive effect of miRNAs on the differentiation of hDP-MSCs and studies evaluating changes to the expression of miRNAs during the differentiation of hDP-MSCs were included. miRNAs involved in odontogenic/osteogenic differentiation were then included in a bioinformatic analysis. A miRNA-mRNA network was constructed, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed. A protein–protein interaction (PPI) network was also constructed. Results: Of 766 initially identified records through database searching, 42 and 36 studies were included in qualitative synthesis and bioinformatic analyses, respectively. Thirteen miRNAs promoted and 17 suppressed odontogenic/osteogenic differentiation of hDP-MSCs. hsa-miR-140-5p, hsa-miR-218 and hsa-miR-143 were more frequently reported suppressing the odontogenic/osteogenic differentiation of hDP-MSCs. hsa-miR-221 and hsa-miR-124 promoted and hsa-miR-140-5p inhibited neuronal differentiation, hsa-miR-26a-5p promoted and hsa-miR-424 suppressed angiogenic differentiation, and hsa-miR-135 and hsa-miR-143 inhibited differentiation within myogenic lineages. A miRNA-mRNA network including 1890 nodes and 2171 edges was constructed. KEGG pathway analysis revealed MAPK, PI3K-Akt and FoxO as key signaling pathways involved in the odontogenic/osteogenic differentiation of hDP-MSCs. Conclusions: The findings of this systematic review support the potential application of the specific miRNAs to regulate the directed differentiation of hDP-MSCs in the field of regenerative therapies

The Potential Mechanism of ZFX Involvement in the Cell Growth

Background:The zinc-finger X linked (ZFX) gene encodes a transcription factor that acts as a regulator of self-renewal of stem cells. Due to the role of ZFX in cell growth, understanding ZFX protein-protein interactions helps to clarify its proper biological functions in signaling pathways. The aim of this study is to define ZFX protein-protein interactions and the role of ZFX in cell growth. Materials and Methods: The PIPs output includes three interacting proteins with ZFX: eukaryotic translation initiation factor 3 subunit I(EIF3I), eukaryotic translation initiation factor 3 subunit G(EIF3G) and protein nuclear pore and COPII coat complex component homolog isoform 3 (SEC13L1). Results: As a cargo and transmembrane protein interacting with Sec13,eIF3I and eIF3G, ZFX mediates cargo sorting in COPII vesicles at ER exit sites. While traveling to cis-Golgi, eIF3I is phosphorylated by the mechanistic target of rapamycin (mTOR). Proteins transport by COPI vesicles to the nucleusouter site layer containing SEC13 via the contribution of microtubules. EIF3G and eIF3I interact with coatomer protein complex subunit beta 2 (COPB2) that helps to enclose ZFX in COPI vesicle. ZFX and eIF3G enter nucleolus where activation of transcription from pre rDNA genes occurs. Conclusion:We proposed a model in which ZFX is involved in cell growth by promoting the transcription of rDNA genes