Muradiye Acar

Relationship between cytosine-adenine repeat polymorphism of ADAMTS9 gene and clinical and radiologic severity of knee osteoarthritis

The aim of this study is to determine the role of cytosine‐adenine (CA) micro‐satellite repeat sequence of ADAMTS9 gene on the development and progression of osteoarthritis (OA).

Matrix Metalloproteinases 2 and 9 Polymorphism in Patients With Myeloproliferative Diseases: A STROBE-Compliant Observational Study

AbstractChronic myeloproliferative disorders such as polycythemia vera (PV), essential thrombocytosis (ET), and idiopathic myelofibrosis arise from clonal proliferation of neoplastic stem cells in the bone marrow. Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that have potential to degrade all types of extracellular matrix (ECM) and also play a role in remodeling of the ECM. It is known that MMPs play a role in bone marrow remodeling.The primary goal of our study is to explore the relationship between chronic myeloproliferative diseases and some of MMP gene polymorphisms. The demonstration of a relationship will help to understand whether these polymorphisms may be a potential early diagnosis marker of the diseases.Patients were selected from outpatient clinics of Turgut Ozal University Hospital, Ankara, Turkey, between December 2010 and May 2011. Twenty-eight patients that previously diagnosed and followed-up with PV, 17 with secondary polycythemia (SP), and 12 with ET were enrolled in the study, along with a control group of 22 healthy people.DNA was isolated from peripheral blood. Using polymerase chain reaction–restriction fragment length polymorphism method, MMP2 and MMP9 gene polymorphisms were analyzed with agarose gel electrophoresis. There was a statistically significant difference between the study groups and the control group in terms of Gln279Arg polymorphisms rates of MMP9. The highest MMP9 Gln279Arg polymorphism rate was observed in the ET group. But nobody from the control group had polymorphic MMP9. There was no statistically significant difference between the groups in terms of MMP2-735 C > T polymorphism rates.In conclusion, MMP9 gene Gln279Arg polymorphism was associated with ET, SP, and PV diseases. Hence, we believe that these gene polymorphisms may play a role in the mechanism of bone marrow fibrosis and may be a factor that increases the risk of thrombosis. Illumination of the molecular basis of the relationship between MMP-thrombosis and MMP-fibrosis provides a better understanding of the pathophysiology of PV and ET diseases and will allow new approaches to diagnosis and treatment.

Is There a Relationship Between Pelvic Organ Prolapse and Tissue Fibrillin-1 Levels?

PURPOSE Pelvic organ prolapse is a multifactorial disorder in which extracellular matrix defects are implicated. Fibrillin-1 level is reduced in stress urinary incontinence. In Marfan syndrome, which is associated with mutations in Fibrillin-1, pelvic floor disorders are commonly observed. We hypothesize that Fibrillin-1 gene expression is altered in pelvic organ prolapse. METHODS Thirty women undergoing colporrhaphy or hysterectomy because of cystocele, rectocele, cystorectocele, or uterine prolapse were assigned to a pelvic prolapse study group, and thirty women undergone hysterectomy for nonpelvic prolapse conditions were assigned to a control group. Real-time polymerase chain reaction was conducted on vaginal tissue samples to measure the expression of Fibrillin-1. Expression levels were compared between study and control groups by Mann-Whitney U test with Bonferroni revision. RESULTS Fibrillin-1 gene expression was not significantly lower in the study group than in the control group. Similarly, no significant correlation between Fibrillin-1 levels and grade of pelvic prolapse was found. Age over 40 years (P=0.018) and menopause (P=0.027) were both associated with reduced Fibrillin-1 levels in the pelvic prolapse group, whereas the delivery of babies weighing over 3,500 g at birth was associated with increased Fibrillin-1 expression (P=0.006). CONCLUSIONS The results did not indicate a significant reduction in Fibrillin-1 gene expression in pelvic prolapse disorders; however, reduced Fibrillin-1 may contribute to increased pelvic organ prolapse risk with age and menopause. Increased Fibrillin-1 gene expression may be a compensatory mechanism in cases of delivery of babies with high birth weight. Further studies are needed for a better understanding of these observations.

Functional analysis of ESM1 by siRNA knockdown in primary and metastatic head and neck cancer cells

BACKGROUND Genetic factors play a large role in cancer, and thus, there is a great desire to understand the effects of different genes in cancer and to also develop gene therapy for better treatments. Therefore, the development of alternative diagnosis and therapy modalities is of utmost importance. The aim of our study was to illuminate the role of ESM1 (endothelial cell-specific molecule-1, also known as Endocan) in proliferation and migration of head and neck cancer, thus helping to pave the way for new treatment modalities and predictive biomarkers. METHODS ESM1 expression was shown with immunofluorescence assay using confocal laser scanning microscope in primary and metastatic head and neck cancer cells. ESM1 expression was knocked down by RNA interference in head and neck cancer cells. Knockdown efficiency was evaluated by quantitative real-time RT-PCR and Western blot. Cell proliferation and migration assays were performed by xCELLigence real-time cell analysis system. RESULTS Immunofluorescence assay showed nuclear localization and high expression of ESM1 in primary and metastatic head and neck cancer cells. ESM1 mRNA and protein levels were significantly decreased in ESM1-knockdown cells compared to control. ESM1-knockdown cells showed reduced proliferation and migration activity when compared to control cells. CONCLUSION These findings suggest that ESM1 has roles on proliferation and migration of head and neck cancer cells.

Abstract 1399: Characterization of cancer stem cell properties and identification of invasion as well as metastatic process in head and neck cancer

Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA Cancer stem cells (CSC) are thought to be a sub-population of cancer cells, which are the main tumorigenic cells in the tumors. Although chemoradiotherapy may kill the majority of rapidly dividing cells, CSC-like cell populations may be left behind due to their low turnover and infrequent cell cycling. In 2007, Prince et al. reported the presence of head and neck cancer stem cells for the first time. In the current study, UTTSCC74A and UTTSCC74B head and neck cancer cell lines as material derived from the primary tumor and their lymph node metastasis, respectively, were used. FACS was used for isolation of cancer stem cells using antibodies against CD44 and ALDH1. However we could not get specific response from CD44 antigen ie can not isolate cancer stem cell. However ALDH1 antibody selection gave us isolation of cancer stem cells from both of UTTSCC74A and B cell lines though much less cancer stem cells existed in UTTSCC74B cell line. The cells were first cultured in normal DMEM with 10% FBS under the condition of 5% CO2. Then the cells were changed into stem cell medium and cultured. Cancer stem cells were isolated using antibody against ALDH1 and these cells were cultured in stem cell medium and its characteristics were confirmed through sphere formation as well as immunofluorescence staining with antibody against ALDH1 and control antibody against DAPI. Expressions of sox-2, oct-4 and klf-4 were confirmed. After characterization of cancer stem cell from head and neck cancer cell line was confirmed, microarray analysis were performed using Gene Chip PrimeView Human expression Array (U133-HG133/ 47,000 transcript). Each group of ALDH1+ (cancer stem) and - cells isolated from each of UTTSCC74A and UTTSCC74B cell lines were exposed to microarray analysis. RNA extraction and cDNA construction from each sub group were prepared for microarray analysis. Comparison of UTTSCC74A ALDH1+ and - provided increased expression of 2037 and decreased expression in 2263 genes in various ratios. Similarly comparison of UTTSCC74B ALDH1+ and - provided increased expression of 6349 and decreased expression in 5322 genes in various ratios. 26 genes with most up and down regulation were selected and confirmation was done through real-time RT-PCR designing custom primers. Similar results were taken as in microarray analysis. From these 26 genes with confirmation of their expressions, 5 up regulated and 5 downregulated genes were selected for further analysis. We currently prepare expression plasmids of these genes and continue their in vitro functional analysis. Thus, the identification of the cancer-stem cell related signalling pathways in head and neck cancer stem cell would provide valuable information for a better understanding of cancer stem cell properties, molecular mechanisms of carcinogenesis, invasion and metastasis in this cancer type. By this way, effective therapeutic options targeting CSCs may be developed. Citation Format: Mehmet Gunduz, Omer Faruk Hatipoglu, Esra Gunduz, Elif Nihan Cetin, Eyyup Uctepe, Sadik Cigdem, Reidar Grenman, Muradiye Acar. Characterization of cancer stem cell properties and identification of invasion as well as metastatic process in head and neck cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1399. doi:10.1158/1538-7445.AM2015-1399

Oncogenes and Tumor Suppressor Genes as a Biomarker in Breast Cancer

Breast cancer is the most common cause of cancer in women in the United States and the Western world. The important question is what can be done to limit the human suffering associated with cancer and to reduce the burden on society? One solution is early detection. Early diagnosis of breast cancer before symptoms emerge is the most effective prevention of breast cancer.