Zrf1’s role in mESC differentiation and breast cancer progression

Zuotin-related factor 1 (Zrf1) has recently been identified as an epigenetic regulator of gene transcription. Upon a differentiation stimulus, Zrf1 facilitates the expression of its target genes via specifically binding to mono-ubiquitinated histone H2A and simultaneous displacement of PRC1 from chromatin. So far, Zrf1 was shown to primarily operate during differentiation into the neuronal lineage. As a multifunctional protein, Zrf1 plays a crucial role in carcinogenesis. Zrf1 is overexpressed in many cancers and was suggested to have an oncogenic role in cancer progression. Zrf1 provides leukemogenic potential in acute myeloid leukemia and it contributes to Ras-oncogene induced senescence through the INK4-ARF locus. In the present study, I elucidated a broader role for Zrf1 during during in vitro differentiation. Zrf1 depleted mESCs have defects in the generation of all three germ layers. I observed a particularly pronounced effect during mesoderm development and I was able to rescue the observed phenotype by re-expression of Zrf1 in Zrf1 knockdown cells. I found that Zrf1 plays a prominent role in cardiomyocyte differentiation and Zrf1 knockdown results in impaired beating function of cardiomyocytes in vitro. At the molecular level, Zrf1 plays an essential regulatory role for the expression of the first and second heart field related genes, thereby promoting faithful cardiomyocyte differentiation. Taken together, the function of Zrf1 needs to be considered when addressing molecular mechanisms of cardiovascular diseases. In addition to studying the function of Zrf1 during differentiation, I have studied the function of Zrf1 in breast invasive ductal carcinoma. Upon Zrf1 depletion, breast cancer cells increase their cell motility, decrease their cellular adhesion and are able to form organoid like structures in cell suspension. Hence, Zrf1 depleted cells can mimic the events during cancer metastasis in vitro. Zrf1 knockdown also contributes to hormone independent growth of cells which represents a phenotype that is observed in more aggressive forms of breast cancer. Zrf1 depletion disrupts the balance between anti-apoptotic and pro-apoptotic genes, thereby favoring cell survival and contributing to drug resistance. My data suggest that Zrf1 is a potential novel target to be explored for new treatment strategies in breast cancer, moreover it plays a critical regulatory role in the progression of breast invasive ductal carcinoma.Zuotin-related factor 1 (Zrf1) wurde vor kurzem als ein epigenetischer Regulator der Transkription beschrieben. Während der Zelldifferenzierung unterstützt Zrf1 die Gen-Expression durch ein spezifisches Binden an mono-ubiquitiniertes Histone H2A und gleichzeitiger Verdrängung der PRC1 Komplexe vom Chromatin. Bislang wurde gezeigt, dass Zrf1 hauptsächlich während der Zelldifferenzierung in die neuronale Linie beteiligt ist. Als ein multi-funktionales Protein spielt Zrf1 auch eine wichtige Rolle während der Krebsentstehung. Zrf1 ist in vielen Krebsarten überexpremiert und es wird vermutet, dass es eine Funktion in der Metastasierung hat. In akuter Leukämie spielt Zrf1 eine signifikante Rolle, und es trägt zur Ras-induzierten Seneszenz durch den INK4A-ARF locus bei. In der hier vorliegenden Arbeit wurde eine globale Analyse der Zrf1 Funktion während der In-vitro-Differenzierung vorgenommen. Zrf1-depletierte embryonale Maus-Stammzellen zeigen Defekte in der Formation aller drei Keimblätter. Es konnte gezeigt werden, dass insbesondere die Entwicklung des Mesoderms in Zrf1-Knockdown Zellen gestört ist. Die Ausbildung dieses Phänotyps konnte nach ektopischer Expression von Zrf1 in den Knockdown Zelllinien verhindert werden. Es konnte weiterhin gezeigt werden, dass Zrf1 eine essentielle Funktion während der Differenzierung von Kardiomyozyten einnimmt. Unter anderem zeigen Zrf1 Knockdown Zellen eine verminderte Schlagrate in vitro. Auf molekularer Ebene spielt Zrf1 eine wichtige regulative Rolle bei der Expression der Gene des ersten und zweiten Herzfelds. Zusammengenommen sollte in Zukunft die Funktion von Zrf1 berücksichtigt werden, wenn molekulare Mechanismen von kardiovaskulären Krankheiten untersucht werden. Weiterhin widmet sich die vorliegende Arbeit der Rolle von Zrf1 in invasivem Brustkrebs. Ein Zrf1 Knockdown in Brustkrebszellen bewirkt eine höhere Zellmobilität, eine verringerte Adhäsion und eine Ausbildung von Organoid-Strukturen. Ein Zrf1 Knockdown führt daher zu einer erhöhten Metastasierung in vitro und trägt ebenfalls zu einem Hormon-unabhängigen Wachstum der Zellen bei, einem Phänotyp den man gewöhnlich in aggresiven Formen des Brustkrebs findet. Eine Depletion von Zrf1 hebt die Balance der Expression von pro-apoptotischen und anti-apoptotischen Genen auf, wodurch es zu einem Überleben der Zellen und Resistenz gegenüber Medikamenten kommen kann. Daher stellt Zrf1 eine neues drug target dar, welches neue Behandlungsstrategien eröffnen könnte

Detection of Marker Chromosome in the Abortion Material; Does It Reflect the Karyotype of the Pregnancy Lost Tissue or the Maternal Decidual Tissue? Case Report

Determination of the origin of the marker chromosomes is very difficult and requires effort. Evaluation of the marker chromosomes of the pregnancy lost tissue could be the most difficult one as the contamination with maternal decidual cells complicates this process. Conventional cytogenetic techniques and fluorescence in situ hybridization technique or more advanced molecular techniques are preferred to determine the origin of these chromosomes. In this report, the study for identification of the origin of an abortion material derived from the primary tissue cell culture, with a 47,XX,+mar(15) karyotype was presented, by using an effective algorithmic approach which ended with genotyping. The result of the genotyping was informative, since the assumed abortion material actually belonged to the mother's decidual tissue. This brief study reminds the efficient algorithmic approaches to the determination process of the marker chromosomes of the abortion material

Optimization of fucoxanthin extraction with different techniques from Phaeodactylum tricornutum

European Biotechnology Congress -- MAY 25-27, 2017 -- Dubrovnik, CROATIAWOS: 000413585400219Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [115O578]The authors acknowledge the Scientific and Technological Research Council of Turkey (TUBITAK) (Project Number: 115O578) for financial support

Analysis of tumor necrosis factor alpha-induced and nuclear factor kappa B-silenced LNCaP prostate cancer cells by RT-qPCR

WOS: 000345948000005Prostate cancer is the second leading cause of morbidity and mortality in males in the Western world. In the present study, LNCaP, which is an androgen receptor-positive and androgen-responsive prostate cancer cell line derived from lymph node metastasis, and DU 145, which is an androgen receptor-negative prostate cancer cell line derived from brain metastasis, were investigated. TNF alpha treatment decreased p105 and p50 expression and R1881 treatment slightly decreased p105 expression but increased p50 expression with or without TNF alpha induction. As an aggressive prostate cancer cell line, DU145 transfected with six transmembrane protein of prostate (STAMP)1 or STAMP2 was also exposed to TNF alpha. Western blotting indicated that transfection with either STAMP gene caused a significant increase in NF kappa B expression following TNF alpha induction. In addition, following the treatment of LNCaP cells with TNF alpha, reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed with a panel of apoptosis-related gene primers. The apoptosis-related genes p53, p73, caspase 7 and caspase 9 showed statistically significant increases in expression levels while the expression levels of MDM2 and STAMP1 decreased following TNF alpha induction. Furthermore, LNCaP cells were transfected with a small interfering NF kappa B (siNF kappa B) construct for 1 and 4 days and induced with TNF alpha for the final 24 h. RT-qPCR amplifications were performed with apoptosis-related gene primers, including p53, caspases and STAMPs. However, no changes in the level of STAMP2 were observed between cells in the presence or absence of TNF alpha induction or between those transfected or not transfected with siNF kappa B; however, the level of STAMP1 was significantly decreased by TNF alpha induction, and significantly increased with siNF kappa B transfection. Silencing of the survival gene NF kappa B caused anti-apoptotic STAMPI expression to increase, which repressed p53, together with MDM2. NF kappa B silencing had varying effects on a panel of cancer regulatory genes. Therefore, the effective inhibition of NF kappa B may be critical in providing a targeted pathway for prostate cancer prevention.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [106S295]; Turkish Academy of Sciences (TUBA)Turkish Academy of Sciences [GEBIP-2007]This study was supported by grants from The Scientific and Technological Research Council of Turkey (TUBITAK) to CGK (Grant no: 106S295) and The Turkish Academy of Sciences (TUBA) to CGK (GEBIP-2007)

The Regulation of Matrix Metalloproteinase Expression and the Role of Discoidin Domain Receptor 1/2 Signalling in Zoledronate-treated PC3 Cells

WOS: 000360298900013PubMed ID: 26366216Discoidin Domain Receptors (DDR1/DDR2) are tyrosine kinase receptors which are activated by collagen. DDR signalling regulates cell migration, proliferation, apoptosis and matrix metalloproteinase (MMP) production. MMPs degrade extracellular matrix (ECM) and play essential role in tumor growth, invasion and metastasis. Nitrogen-containing bisphosphonates (N-BPs) which strongly inhibit osteoclastic activity are commonly used for osteoporosis treatment. They also have MMP inhibitory effect. In this study, we aimed to investigate the effects of zoledronate in PC3 cells and the possible role of DDR signalling and downstream pathways in these inhibitory effects. We studied messenger RNA (mRNA) and protein expressions of MMP-2,-9,-8, DDR1/DDR2 type I procollagen (TIP) and mRNA levels of PCA-1, MMP-13 and DDR-initiated signalling pathway players including K-Ras oncogene, ERK1, JNK1, p38, AKT-1 and BCLX in PC3 cells in the presence or absence of zoledronate (10-100 mu M) for 2-3 days. Zoledronate (100 mu M) down-regulated DDR1/DDR2, TIP mRNAs but did not change MMP-13 (collagenase-3) mRNA. However, zoledronate up-regulated MMP-8 (collagenase-2) mRNA. Zoledronate also inhibited mRNA expressions of K-Ras, ERK1, AKT-1, BCLX and PCA-1; but did not change JNK1, p38 mRNA levels. Zoledronate (100 mu M) supressed DDR1/DDR2, TIP expressions; and gelatinase (MMP-2/MMP-9) expressions/activities. Conversely, zoledronate up-regulated MMP-8 expression in PC3 cells. Zoledronate down-regulates MMP-2/-9 expressions in PC3 prostate cancer cells. DDR1/DDR2 signalling and DDR-initiated downstream Ras/Raf/ERK and PI3K/AKT pathways may at least partially responsible for MMP inhibitory effect of zoledronate.Scientific Research Foundation of Ege University, Izmir, TurkeyEge University [13/ECZ/012]This research was supported by the Scientific Research Foundation of Ege University, Izmir, Turkey [13/ECZ/012 to Buket Reel]

Introducing Basic Molecular Biology to Turkish Rural and Urban Primary School Children Via Hands-on PCR and Gel Electrophoresis Activities

WOS: 000333236800003PubMed ID: 24474053This study includes the results of a 2-day education project titled Molecular Biology Laboratory Summer School, MoBiLYO. The project was held at a University Research Center by scientists from Department of Pharmacology and graduate students. The project was composed of introductory lectures, model construction, DNA isolation, polymerase chain reaction (PCR), and gel electrophoresis. The participants were 13-year-old eighth-graders attending primary schools affiliated with Ministry of National Education in urban and rural areas of Izmir, Turkey. The purpose of this study was to introduce basic molecular biology concepts through individually performed experiments such as PCR and gel electrophoresis integrated with creative drama. The students were assessed at the beginning and the end of each project day via mini-tests, experimental and presentation skills evaluation forms. Data showed that students' knowledge about DNA structure and basic molecular biology techniques significantly increased. On the basis of experimental and presentational skills, there was no significant difference between kids from urban and rural schools or between public and boarding public schools, whereas the average score of girls was significantly higher than that of boys. In conclusion, individually performed experiments integrated with creative drama significantly increased students' perception of complex experimental procedures on basic molecular biology concepts. Data suggests that integration of these concepts into the science and technology curriculum of Turkish primary education may support the recruitment of future scientists who can handle rapidly developing genomic techniques that will affect our everyday life. (c) 2014 by The International Union of Biochemistry and Molecular Biology, 42(2):114-120, 2014Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [112B052]; GENOSIS Biotechnology; State Planning Organization Infrastructure ProjectTurkiye Cumhuriyeti Kalkinma Bakanligi [2009K120640]This work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Science and Society Projects, 112B052 to CS) and performed in collaboration with the Ministry of National Education and the Ministry of Family and Social Policies, Izmir, Turkey. A partial support was also provided by the GENOSIS Biotechnology. Project activities took place in Molecular Biology Laboratory (MoBiL) of Pharmaceutical Sciences Research Laboratory (FABAL) constructed with the support of the State Planning Organization Infrastructure Project (2009K120640 to Faculty of Pharmacy, Ege University)

Burden of Chronic Low Back Pain in the Turkish Population

WOS: 000353480500011Objective: Chronic low back pain (CLBP) is a great economic burden to the society mainly in terms of the large number of the lost work days and disability, and it appears to be growing. The economic burden of LBP in Turkey is not known. This study aims to analyze the health care resource use, work and productivity loss, and health-related economics of CLBP in Turkey. Material and Methods: The study was designed as a multi-centered cross-sectional survey of patients in physical therapy and rehabilitation clinics from eight different regions of Turkey and 662 patients with CLBP over 18 years of age were included. Data on patient sociodemographics, disease-related healthcare resource use during the previous 6 months, inability to work during the last 3 months, Roland Morris Disability Index for the functional status, and psychological health with Beck Depression Scale were collected. Direct costs included medical visits, investigations, medications, hospitalizations, orthopedic aids, and physical therapy. Indirect costs were evaluated mostly with productivity loss. Results: The total annual direct costs for CLBP per patient were estimated at 1080 TL. The indirect costs were estimated at 5511 TL per patient. Direct cost was correlated with disease severity, duration, and age. Indirect cost was higher in women. Conclusion: The indirect costs for CLBP were significantly higher than the direct costs