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Amaç: Bu çalışmanın amacı, sıçanlarda maternal uterin arter ligasyonuna bağlı olarak gelişen intrauterine büyüme geriliğinde kondrogenez ve osteogenezin Wnt/β-catenin ve S100 ekspresyonlarının immünohistokimyasal ve histomorfometrik olarak değerlendirilmesi.Gereç ve Yöntem: Gebe sıçanlar 3 gruba ayrıldı (n=5). Kontrol grubuna cerrahi veya anestezi uygulanmadı. Deney grubunda, 18. gebelik gününde bilateral maternal uterin arterlere ligasyon yapıldı. Sham grubuna uterin arter ligasyonu dışındaki tüm cerrahi işlem uygulandı. Fetuslar gebeliğin 20. gününde alındı, sol proksimal tibiadan büyüme plağı ve zonların kalınlıkları, trabeküler sayı ve kalınlıkları, kortikal kalınlıklar histomorfometrik olarak değerlendirildi. Büyüme plağında β-catenin ve S100 ekspresyonları immunohistokimyasal olarak değerlendirildi.Bulgular: Deney grubunda büyüme plağı (p0.05). Aynı zamanda daha düşük sayıda trabekülar yapı (p, Aim: The aim of this study is to investigate the effects of intrauterine growth retardation depending on maternal uterine artery ligation, Wnt/β-catenin and S100 expression immunohistochemistry and histomorphometrically on chondrogenesis and osteogenesis of fetal rats.Materials and Methods: Maternal rats were randomly divided into 3 groups (n=5). No surgery or anesthesia were applied in control group. Bilaterally the maternal uterine arteries were ligated on gestational day 18 in experimental group. Although all surgical procedures were performed in sham group, the uterine artery ligation were not made. Fetuses were taken on gestational day 20, thicknesses of growth plate and zones, trabecular number and thickness and cortical thickness were evaluated with the histomorphometrically in samples from left proximal tibia. The expressions of β-catenin and S100 immunohistochemically were evaluated in the growth plate.Results: Thicknesses of growth plate (p0.05). Also trabecular numbers were lower (p

Aims: The aim of this study was to investigate the cellular binding site of human KDN (2-keto-3-deoxy-D-glycero-D-galacto-nononic acid). The KDN molecule is a member of the sialic acid family, and its expression increases in cancer cells. Although KDN has been shown to bind to GM3 (Monosialodihexosyl Ganglioside) in trout sperm.Methods and Results: Prostate cancer cell line (DU145) was used in this study. Each experimental group was divided into 3 groups: control, GCS (Glucosylceramide synthase) enzyme inhibitor Genz-123346 treated, and GM3 synthesis inhibitor Triptolide treated. Each group was stained using the immunocytochemical method for GM3, GD3 (Disialosyllactosylceramide) and KDN. The FTIR (Fourier Transform Infrared Spectroscopy) analysis was performed to elucidate the cellular changes upon treatment. The non-treated number 1 cell group stained positive with all of GM3, GD3 and KDN, the GCS enzyme blocked with Genz-123346 number 2 cell groups stained positive with only KDN. Furthermore, GD3 Synthase inhibitor Triptolide treated number 3 cell group stained positive with GM3 and KDN. Measurements with FTIR showed apoptotic features with Triptolide while Genz-123346 had no negative effect on the cell viability. The decrease in sugar constructions was revealed and the results that we obtained with staining were reinforced.Conclusions: Determining the location of bounded KDN is important in selecting new targets for cancer treatment researches. It has been shown that KDN is not inhibited by both GM3 inhibition and GD3 inhibition, and thus, KDN might also bind to different places, be specific, and not only attached to any of gangliosides of the GM or GD series.

The current cancer studies focus on the molecular-targeting diagnostics and their interaction with surrounding microenvironment, however, there are some missing points on the characterization of the cells with their topological differences and elemental composition. Glioblastoma multiforme (GBM) which is an astrocytic aggressive brain tumour with short survival time. GBM and astrocyte cells may differ at molecular level and the elemental and topological evaluation of these cells are vital for a definition of new potential targets for cancer research. Here, we report the topology and chemistry of cancer (GBM) and healthy (astrocyte) cells by atomic force microscopy (AFM), scanning electron microscopy (SEM) supported with energy dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS), for the first time. Additionally, F-actin Immunofluorescence staining and Real-Time Polymerase chain reaction (RT-PCR) techniques were performed. Actin related genes were similar in level of gene expression; however, F-actin protein intensities were different. The gene expressions related to the invasion were elevated in GBM cells. Morphologically, GBM cells were found to be longer and narrower while astrocytes were shorter and more disseminated based on AFM. Furthermore, roughness values of these cells were relatively close to each other. SEM-EDS analysis demonstrated that elongated GBM cells exhibited several filopodial protrusions whereas the astrocyte surfaces were rougher in lamellipodial area. Our investigation provides considerable further insight into rapid cancer cell characterization field in terms of its combinatorial spectroscopic and microscopic approach.

Amaç: Çalışmamızda, insan prostat kanser hücre hattı (DU145) ve prostat normal epitel hücre hatları (RWPE) arasında miRNA ifadesinin analizini yapmak ve kanser gelişiminde olası rolünü incelemeği amaçladık. Gereçler ve Yöntem: İnsan prostat epitel hücre hattı (RWPE) ve prostat kanseri hücre hatları (DU-145) Amerikan Tip Kültür Koleksiyonu (ATCC)’den temin edildi. Hücre hatlarının çoğaltılmasında ve sürdürülmesinde RPMI 1640 besi ortamı kullanıldı. Transkriptom analizi için RNA izolasyonu yapılarak, kütüphane oluşturuldu, kütüphanenin kantitasyonunun ardından NextSeq500 (illumina) ile sekanslama yapıldı. Dizileme, haritalandırma, bağıl gen ifade ölcümleri gibi biyoinformatik analizler Genomics Workbench v 8 yazılımı kullanılarak GRCh38 referans sekansı ile yapıldı. Bulgular: RWPE normal prostat epitel hücre kültürleri ile DU145 prostat kanser hücreleri karşılaştırıldığı zaman DU145 prostat kanser hücre kültürlerinde, miRNA (hsa-mir-8072) ifadesinde anlamlı bir artma (p, Objective: in this study, we aimed to analyze miRNA expression between prostate cancer cell line (DU145) and prostate normal epithelial cell lines (RWPE) and to investigate its possible role in cancer development. Material and Methods: Human prostate epithelial cell line (RWPE) and prostate cancer cell line (DU145) were acquired from ATCC. Both cell lines were maintanied in RPMI 1640 medium. Total RNA were isolated and fragmented. Adapters were ligated to prepare RNA library for whole trasncriptome experiments. Statistics and bioinformatics analysis including mapping, clustering, sequencing were done by using Genomics Workbench v 8. software. Results: As we compared the normal prostate ephitalial cells (RWPE) and prostate cancer cells (DU 145); miRNA (hsamir-8072) were significantly (p

Soykam, Huseyin Okan/0000-0002-8955-2412, WOS: 000488246200005, PubMed: 31181875, Objective: This study was performed to explore the possibility that each oocyte and its surrounding cumulus cells might have different genetic expression patterns that could affect human reproduction. Methods: Differential gene expression analysis was performed for 10 clusters of cumulus cells obtained from 10 cumulus-oocyte complexes from 10 patients. Same procedures related to oocyte maturation, microinjection, and microarray analyses were performed for each group of cumulus cells. Two differential gene expression analyses were performed: one for the outcome of clinical pregnancy and one for the outcome of live birth. Results: Significant genes resulting from these analyses were selected and the top 20 affected pathways in each group were analyzed. Circadian entrainment is determined to be the most affected pathway for clinical pregnancy, and proteoglycans in cancer pathway is the most affected pathway for live birth. Circadian entrainment is also amongst the 12 pathways that are found to be in top 20 affected pathways for both outcomes, and has both lowest p-value and highest number of times found count. Conclusion: Although further confirmatory studies are necessary, findings of this study suggest that these pathways, especially circadian entrainment in cumulus cells, may be essential for embryo development and pregnancy., University Scientific Research Project Fund (EgeBAP, 2011) [TIP-00014], This work was supported by the University Scientific Research Project Fund (EgeBAP TIP-00014, 2011).

Parkinson's disease (PD) is the second most common neurodegenerative disorder, and the main cause of PD is still not known. Until now, no cure for Parkinson's disease is yet in sight. Caffeic acid phenethyl ester (CAPE) is a polyphenolic component of the propolis, which can be derived from honeybee hive propolis. We aimed to determine the effect of intrastriatal CAPE administration as a neuroprotective agent on 6-hydroxydopamine (6-OHDA)-induced PD model. Adult male Wistar rats weighing 280-320 g were used. The PD model was induced with unilateral intrastriatal 6-OHDA injection. Treatment groups received 20 mu mol/5 mu L/4 day and 80 mu mol/5 mu L/4 day CAPE 24 h after 6-OHDA injection. Eight days after 6-OHDA application, behavioral studies (adhesive tape removal test, open-field test, cylinder test, and apomorphine-induced asymmetric rotational behavior) were performed once more to compare the effects of CAPE on behavior tests. Striatal histological verifications, immunohistochemistry, and stereological quantitation were performed. Our results for the first time showed that, besides improving the motor performance, CAPE treatment also prevents 6-OHDA-induced loss of TH-positive neurons. From our results, CAPE may be a promising clinical agent in the treatment of PD., Scientific Research Projects Coordination Unit of the University of NE [151218009], This study was supported by grants from Scientific Research Projects Coordination Unit of the University of NE, project no. 151218009.

Objective:Cancer stem cells (CSCs) are a small population in cancer, which are responsible for therapeutic resistance, relapse and metastasis. Flavopiridol has antitumor activity against various types of cancer cells. The mechanism of action of flavopiridol on CD44+/CD24- breast CSCs has not yet been fully elucidated. The aim of this study was to evaluate the mechanism of action of flavopiridol on breast CSCs (BCSC) in terms of apoptosis, cell cycle and biomolecular changes.Methods:In human breast cancer, cells with CD44+/CD24− markers were isolated from MCF-7 cell line using flow cytometry. The induction of apoptosis was investigated by Annexin-V. The effect of flavopiridol on cell cycle arrest was determined and the percent of cell populations at G0/G1, S and G2/M cycles were identified. The effect of the drug on three-dimensional cell cultures was investigated using a multicellular tumor spheroid model. In addition, the effect of flavopiridol on biomolecules has been evaluated using Fourier transform infrared (FTIR) spectroscopy, which has recently been used effectively in various scientific fields.Results:Flavopiridol especially induced early apoptosis. Cell cycle analyses revealed that flavopiridol induced cell cycle arrest in G0/G1 phase. Decreased number and diameter of spheroids was observed following flavopiridol treatment. ATR-FTIR data showed that treatment with flavopiridol led to significant changes in nucleic acids.Conclusion:According to the data obtained in this study, flavopiridol exhibits anticancer effects by altering the structure/ expression level of nucleic acids and changing cell cycle progression and inducing apoptosis. These finding reveals that flavopiridol can be an effective antitumor agent for the treatment of breast cancer after in vivo and phase studies are completed.

EgeUn###, Current cancer treatments destroy the tumor mass but cannot prevent the recurrence of cancer. The heterogeneous structure of the tumor mass includes cancer stem cells that are responsible for tumor relapse, treatment resistance, invasion and metastasis. The biology of these cells is still not fully understood; therefore, effective treatments cannot be developed sufficiently. Herein, attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy, combined with unsupervised multivariate analysis, was applied to prostate cancer stem cells (CSCs), non-stem cancer cells (non-CSCs) and normal prostate epithelial cells to elucidate the molecular mechanisms and features of CSCs, which are crucial to improving the target specific therapies. This work revealed the spectral differences in the cellular mechanisms and biochemical structures among three different cell types. Particularly, prostate CSCs exhibit differences in the lipid composition and dynamics when compared to other cell types. CSCs also harbor pronounced differences in their major cellular macromolecules, including differences in the protein amount and content (mainly ?-helices), the abundance of nucleic acids (DNA/RNA), altered nucleic acid conformation and carbohydrate composition. Interestingly, macromolecules containing the CO groups and negatively charged molecules having the COO- groups are abundant in prostate CSCs in comparison to prostate non-CSCs and normal prostate cells. Overall, this study demonstrates the potential use of ATR-FTIR spectroscopy as a powerful tool to obtain new insights into the understanding of the CSC features, which may provide new strategies for cancer treatment by selectively targeting the CSCs. © 2019 The Royal Society of Chemistry.

Amaç: Prostat kanser hücre hattı (DU145) ve prostat normal epitel hücre hatları (RWPE) arasında anoikis mekanizmasını arttıracak veya inhibe edebilecek genlerin analizini yapmak ve kanser gelişiminde olası rolünü incelemek. Gereç ve Yöntem: İnsan prostat epitel hücre hattı (RWPE) ve prostat kanseri hücre hatları (DU-145) Amerikan Tip Kültür Koleksiyonu (ATCC)’den temin edildi. Hücre hatlarının çoğaltılmasında ve sürdürülmesinde RPMI 1640 (Biological Industries) besi ortamı kullanıldı. Transkriptom analizi için RNA izolasyonu yapılarak, kütüphane oluşturuldu, kütüphanenin kantitasyonunun ardından NextSeq500 (illumina) ile sekanslama yapıldı. Dizileme, haritalandırma, bağıl gen ifadeleri ölçümleri gibi biyoinformatik analizler Genomics Workbench v 8 (Qiagen) yazılımı kullanılarak GRCh38 referans sekansı ile yapılmıştır. Bulgular: RWPE Normal prostat epitel hücre kültürleri ile DU145 prostat kanser hücreleri karşılaştırıldığı zaman DU-145 prostat kanser hücre kültürlerinde, ZNF304, PYCARD ve Notch3 gen expresyonlarında anlamlı bir artış (p

Background/aims Cholestasis, which results in hepatic cell death, fibrosis, cirrhosis, and eventually liver failure, is associated with oxidative stress. The aim of this study was to evaluate the effects of milk thistle (MT, Silybum marianum) and ursodeoxycholic acid (UDCA) or their combination on the activation of hepatic stem cells and on the severity of cholestasis liver injury in rats. Materials and methods Under anesthesia, bile ducts of female Sprague Dawley rats were ligated (BDL) or had sham operation. BDL rats were administered saline, UDCA (15 mg/kg/d), MT (600 mg/kg/d), or UDCA+MT by gavage for 10 days. On the 11th day, rats were sacrificed and blood and liver samples were obtained. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hepatic malondialdehyde (MDA) levels, and myeloperoxidase (MPO) activity were measured. Hepatic injury, a-smooth muscle actin expression, and stem cell markers c-kit, c-Myc, Oct3/4, and SSEA-1 were histologically determined. Results Histological scores, serum ALT, and hepatic MDA levels were higher in BDL group than in the sham rats, while all treatments significantly reduced these levels. The reduction in ALT was significantly greater in UCDA+MT-treated group than in other treatment groups. c-Kit, c-Myc, Oct3/4, and SSEA-1 were increased in saline-treated BDL group with respect to sham-operated control group, and these markers were significantly reduced in all treatment groups. Conclusion In addition to a modulatory effect on the stem cell-induced regenerative response of the liver, UDCA, MT, and their combination demonstrated similar anti-inflammatory and antiproliferative effects on cholestasis-induced hepatic injury.

Mammalian target of rapamycin (mTOR) is a member of the PI3K/Akt/mTOR signaling pathway that participates in cell growth, proliferation, protein synthesis, transcription, angiogenesis, apoptosis and autophagy. We investigated the role of mTOR and other signaling molecules in the rat uterus during implantation. Female pregnant rats were divided into three groups: embryonic days (ED) 4.5, 5.5 and 6.5 according to vaginal smears. Immunohistochemical staining of mTORC1, mTORC2, IGF1, PI3K, pAkt1/2/3, ERK1 and pERK1/2 was performed on formalin fixed, paraffin embedded uterine tissue samples. pAkt1/2/3 and ERK1 also were analyzed using western blotting. We found that PI3K/Akt/mTOR and ERK/pERK were increased during the implantation period. Different amounts of mTORC1, mTORC2, IGF1, PI3K, pAKT1/2/3, ERK1 and pERK1/2 were expressed in luminal epithelium, decidual cells, embryoblast and trophoblast cells during implantation. We suggest that mTOR and associated signaling molecules may participate in implantation. Celal Bayar Üniversitesi Bilimsel Araştırma Proje Komitesi - 2011/038

Lung cancer is one of the most commonly diagnosed cancers and it is associated with high rates of morbidity and mortality. Metastasis and relapse of the tumor depend on the survival and proliferation of lung cancer stem cells (LCSCs). The ability to identify CSCs may prevent recurrence and lead to more effective treatments. Sirtuins are a group of deacetylases that include seven variants (SIRT1-7), with sirtuin 1 (SIRT1) being the most intensively investigated. Evidence suggests that SIRT1 is both a tumor-suppressor gene and an oncogene. SIRT1 can deacetylate the tumor-suppressor protein p53 to decrease its activity. SIRT1 activators increase the deacetylation of p53, whereas SIRT1 inhibitors can stimulate p53 by inhibiting deacetylation. In the present study, CD44+ and CD133+-enriched A549 (non-small cell lung cancer) cells collected using the CD44 and CD133 CSC surface markers by fluorescence?activated cell sorting method were treated with SIRT1 inhibitors (tenovin-6 and sirtinol) and SIRT1 activators (resveratrol and SRT1720), and their effects on apoptosis, as well as the mRNA and protein expression of SIRT1 and p53 were investigated. Of these agents, it was found that resveratrol increased p53 expression by 4.1-fold, decreased SIRT1 expression by 0.2-fold, and it was the most potent inducer of apoptosis. © 2020 Spandidos Publications. All rights reserved., Ege Ãœniversitesi: 15-TIP-061, The current study was supported by the Scientific Research Projects Coordination Unit of Ege University (grant no. 15-TIP-061).

Amaç: Kanser kök hücreleri (KKH), tümör içinde kendi kendilerini yenileme ve diğer hücre tiplerine farklılaşabilme kapasitesi sebebiyle tümörün başlaması, ilerlemesi, nüksetmesi, metastaz ve terapötik dirence yol açmaktadır. Bu nedenle, meme kanser kök hücrelerinin (MKKH) karakteristik özelliklerinin belirlenmesi gerekmektedir. Bu çalışmanın amacı, MKKH?lerin akış sitometrisi ile izole edildikten sonra Fourier dönüşümlü kızılötesi (FTIR) spektroskopisi kullanarak hücre biyokimyasındaki farklılaşmalarının moleküler seviyede araştırılmasıdır. Gereç ve Yöntem: MCF-7 meme kanser hücre hattındaki CD44+/CD24- yüzey belirteç özelliği gösteren MKKH?ler akış sitometrisi ile izole edilmiştir. MCF10A, MCF-7 kanser hücre (KH) hattı ve bu hattan izole edilen CD44+/CD24- yüzey belirteç özelliklerine sahip MKKH?'ler %0,9 NaCI içerisine resuspanse edildikten sonra FTIR spektrometre ile ölçülmüştür. Bulgular: MCF-7 içerisindeki CD44+/CD24- yüzey belirteç özelliğine sahip KKH?lerinin sort oranı %2,0-2,3 olarak belirlenmiştir. Elde edilen FTIR spektrumlarında, MKKH, meme kanser hücreleri (KH, non-KKH, bulk populasyon) ve sağlıklı hücreler arasında spektral benzerlikler ve farklılıklar tespit edilmiştir. MKKH?lerde lipit ve protein sinyalleri daha güçlü olup hücre zarı akışkanlığı ve dinamiği fazladır. Sağlıklı hücreler ile kıyaslandığında, KH?lerde ?-helikal proteinler ve DNA sinyallerinde azalmaya karşın negatif yüklü karboksil gruplarından kaynaklanan sinyallerde artış gözlenmektedir. Bu veriler, MKKH?lerin, sağlıklı ve KH?lere kıyasla yapı, içerik ve dinamiği bakımından oldukça farklı bir profil sergilediğini göstermektedir. Sonuç: Bu çalışma, MKKH?lerinin moleküler yapısı ve içeriğindeki değişikliklerin incelemesi vasıtasıyla terapötik hedefli ilaç çalışmaları yapılabileceğini ortaya koymaktadır. FTIR spektroskopisi boyar madde gerektirmeden, hassas ve hızlı ölçüm alınması, örnek hazırlamada kolaylık ve az miktarda örnek gerektirmesi sebebiyle ileri hücre çalışmalarında ve medikal alanda biyolojik örneklerin analizlerinde kullanılabileceği de gösterilmiştir., Aim: Cancer stem cells (CSCs) lead to tumor initiation, progression, relapse, metastasis and therapeutic resistance due to the ability of tumor to self-renewal and differentiate into other cell types. Therefore, the characteristic features of breast CSCs need to be determined in detail. The aim of this study was to investigate the differences in cell biochemistry at the molecular level by using Fourier transform infrared (FTIR) spectroscopy after breast CSCs were isolated with flow cytometry. Materials and Methods: Breast CSCs with CD44+/CD24- surface marker properties in the MCF-7 cancer cell lines were isolated by using flow cytometry (FACS). MCF10A, MCF-7 breast cancer cell line (cancer non-stem cells or non-CSCs) and breast CSCs were re-suspended into 0.9% NaCl, and each cell type was measured with the FTIR spectrometer. Results: The portion of breast CSCs with CD44+/CD24- surface marker properties in MCF-7 was 2.0- 2.3%. In the FTIR spectra, spectral similarities and differences among breast CSCs, non-CSCs and healthy cells were determined. In breast CSCs, the lipid and protein signals are quite strong accompanied with an increased cell membrane fluidity and dynamics. When non-CSCs are compared with healthy cells, a less amount of both ?-helical proteins and DNA is detected while an increase in the signals of negatively charged carboxyl groups is noticed. These data clearly show that breast CSCs exhibit a very different profile in terms of structure, content and dynamics of cellular macromolecules compared to both non-CSCs and healthy cells. Conclusion: Drug studies (targeted therapy, drug-action mechanism) can be performed by examining small changes in the molecular structure and content of breast CSCs. This study shows that FTIR spectroscopy can be used in advanced cell studies as well as in the analysis of biological samples in medical field due to rapid, label-free and accurate measurement without complex sample preparation procedures.

Background: The members of the transforming growth factor-B superfamily, as the bone morphogenetic proteins (BMPs) subfamily and anti-Müllerian hormone (AMH), play a role during follicular development, and the bone morphogenetic protein-2 (BMP2), AMH, and THY1 are expressed in ovaries. Aim: This study was designed to define whether or not the expressions of these proteins in human cumulus cells (CCs) can be used as predictors of the oocyte and embryo competence. Settings and Design: The study included nine female patients who were diagnosed as idiopathic infertility, aged 25-33 years (median 30 years) and underwent Assisted Reproductive Technologies. Materials and Methods: The CCs from 60 oocyte-cumulus complexes obtained from the nine patients were evaluated with immunofluorescence staining in respect of BMPs, AMH and THY1 markers. The CCs surrounding the same oocytes were evaluated separately according to the oocyte and embryo quality. Statistical Analysis: Quantitative data were statistically analyzed for differences using the two-sided Mann-Whitney U test (P < 0.05). Results and Conclusions: Significant differences in immunofluorescence staining were observed in oocyte quality and embryo quality for the BMP2 only (P < 0.05). No significant differences were observed for AMH or CD90/THY1. Conclusion: These results demonstrated that there is a significant difference in the expression of BMP2 in the CCs of good quality oocytes and subsequently a good embryo. © 2017 Journal of Human Reproductive Sciences Published by Wolters Kluwer - Medknow., Ege-BAP 2011 Tip-00014, This study was approved by the Ethics Committee for Human Research of Ege University (11-2, 9; 28.03.2011). -- The authors thank the financial support rendered by the Ege University Scientific Research Project Fund (Ege-BAP 2011 Tip-00014).

WOS: 000385579200110, PubMed ID: 27703531, Prostate cancer (PCa) is the second most frequent type of cancer in men worldwide and the levels of differentiation growth factor midkine (MK) are increased in PCa. Cancer and/or the treatment process itself may lead to psychiatric disorders. Lithium chloride (LiCl) has anti-manic properties and has been used in cancer therapy; however, it has a queried safety profile. In addition, cancer stem cells are responsible for the heterogeneous phenotype of tumor cells; they are involved in progression, metastasis, recurrence and therapy resistance in various cancer types. The aims of the present study were to investigate the effect of different concentrations of LiCl on PCa stem cells (whether a shift from tumorigenic to non-tumorigenic cells occurs) and to determine if these results can be explained through changes in MK levels. Monolayer and spheroid cultures of human prostate stem cells and non-stem cells were incubated with low (1, 10 mu M) and high (100, 500 mu M) concentrations of LiCl for 72 h. Cell proliferation, apoptotic indices, MK levels and ultrastructure were evaluated. Cells stimulated with low concentrations showed high proliferation, low apoptotic indices, high MK levels and more healthy ultrastructure. Opposite results were obtained at high concentrations. Furthermore, stem cells were more sensitive to stimulation and more resistant to inhibition than non-stem cells. LiCl exhibited concentration-dependent effects on stem cell and non-stem cell groups. MK levels were not involved in the biphasic effect of LiCl; however, they were proportionally affected. To the best of our knowledge, the present study was the first to show the effect of LiCl on PCa stem cells through MK.

EgeUn###, Cancer, which causes the deaths of millions of people, is an important public health problem worldwide. Despite significant advances in the diagnosis and treatment of cancer, survival rates are still insufficient in progressive cancers. Today, one of the most important reasons for not being able to reach the desired level in the fight against progressive cancer types is cancer stem cells (CSCs). The ineffectiveness of conventional therapies on CSCs has made it necessary to investigate the molecular mechanisms and signaling pathways used in the survival, drug resistance, and metastasis of CSCs. In this context, studies investigating the biology of CSCs suggest that lipid metabolism and extracellular vesicles are critical for understanding the stemness and metastasis of these cells. These studies have demonstrated that a number of molecules play a vital role in resistant to apoptosis in CSCs, including the cellular FLICE-inhibitor protein (c-FLIP), which inhibits TRAIL-induced extrinsic apoptosis. Another important output of these studies is the demonstration of the relationship of the cancer microenvironment in terms of epithelial-mesenchymal transition, which CSCs frequently use in the metastasis process. In addition, studies investigating the differences in glycosylation observed in CSCs and investigating cancer vaccines are promisng. These findings will strengthen our aresnal in the fight against cancer. In this article, we summarize current molecular studies on CSCs, an important target in novel cancer therapies. © 2019 Begell House, Inc.

INTRODUCTION: Spheroid cultures reflect properties of tumor tissue better than monolayer cultures in terms of internal dynamics and interaction with other cells. The aim of this study is to investigate the similarities and differences in CD133+/CD44+ prostate cancer stem cells (CSCs) produced in three-dimensional (3D) and two-dimensional (2D) culture media.METHODS: CSCs with CD133+/CD44+ surface marker properties in the DU-145 prostate cancer cell lines were isolated using flow cytometry (FACS). Spheroid structures were formed with agar-coated culture vessels. CSCs in 2D and 3D cultures were compared with Fourier transform infrared (FTIR) spectroscopy.RESULTS: CD133+/CD44+ cells were observed to form micro-aggregates in cultured media in the first week. In the second week, mature spheroid structures were formed. FTIR analysis revealed that the 2D and 3D (multicellular tumor spheroids) models of CSCs exhibit significant differences in proteins, lipids and nucleic acids. Significant differences were detected in membrane lipid acyl chain length, membrane thickness, protein secondary structures and DNA oligonucleotides. The results showed that spheroids in 3D culture medium exhibit similar properties to in vivo tumor tissue.DISCUSSION AND CONCLUSION: Physical, chemical and biological properties generated by environmental conditions significantly affect internal dynamics of cells and their interactions within the microenvironment. Reducing the cells in one dimension through the 2D culture medium does not reflect actual properties of cells. Therefore, cell dynamics in 3D culture media should be investigated. This study demonstrates that cellular lipids, membrane structure, protein secondary structures and nucleic acids of CSCs constituting an important cell population in cancer may be therapeutic targets., GİRİŞ ve AMAÇ: Sferoid kültürleri, hücrelerin kendi iç dinamikleri ve diğer hücrelerle olan etkileşimleri açısından tek tabakalı kültürlere kıyasla tümör dokusunun özelliklerini daha iyi yansıtmaktadır. Bu çalışmanın amacı, üç boyutlu (3D) ve iki boyutlu (2D) kültür ortamlarında üretilen CD133+/CD44+ prostat kanser kök hücrelerinin (KKH) makromoleküllerindeki benzerlik ve farklılıklarının araştırılmasıdır.YÖNTEM ve GEREÇLER: DU-145 prostat kanser hücre hattı içerisindeki CD133+/CD44+ yüzey belirteç özelliklerine sahip KKH’leri akış sitometrisi (FACS) kullanılarak izole edilmiştir. Agarla kaplı kültür kapları ile sferoid yapıları oluşturulmuştur. 2D ve 3D kültür ortamlarındaki KHK hücreleri Fourier dönüşümü kızılötesi (FTIR) spektroskopisi ile karşılaştırılmıştır. BULGULAR: CD133+/CD44+ hücrelerin birinci haftada agarlı kültür ortamlarında mikro-agregatlar oluşturduğu gözlenmiştir. İkinci haftada ise, olgun sferoid yapıların oluştuğu saptanmıştır. 2D ve 3D (multisellüler tümör sferoidleri) kültür ortamlarında üretilen hücreler ile yapılan FTIR analizleri KHK hücre yapısındaki proteinler, lipitler, karbonhidratlar ve nükleik asitlerde (DNA, RNA) önemli derecede farklanmalar olduğunu göstermiştir. Membran lipit açil zincir uzunluğu ve hücre zarı kalınlığı, proteinlerin sekonder yapıları ve DNA oligonükleotitlerin baz sekanslarında veya fonksiyonel gruplarında önemli farklanmaların olduğu tespit edilmiştir. Elde edilen sonuçlar, 3D kültür ortamında üretilen sfreoid yapılarının in vivo tümör dokusu ile benzer özellikler sergilediğini göstermiştir.TARTIŞMA ve SONUÇ: Hücre ortam koşulları ile yaratılmış olan fiziksel, kimyasal ve biyolojik özellikler hücrelerin kendi iç dinamiğini ve mikroçevresi içerisindeki etkileşimlerini önemli derecede etkilemektedir. 2D kültür ortamları ile hücreleri tek boyutta indirgemek hücrelerin gerçek özelliklerini yansıtmamaktadır. Bu nedenle, 3D kültür ortamları ile hücre dinamiklerinin incelenmesi gerekmektedir. Bu çalışmadan elde edilen sonuçlar, kanserde önemli bir hücre popülasyonunu oluşturan KKH’lerin membran yapısı, lipitler, proteinlerin sekonder yapıları ve DNA oligonükleotit yapılarının terapötik hedef olabileceğini göstermektedir.

Aims To examine the functions of growth factor midkine (MK) and a flavonoid quercetin on survival, apoptosis and migration of prostate cancer (PCa) stem cells (CSCs). Main methods CD44+/CD133+ and CD44+ stem cells were isolated from PC3 and LNCaP cells, respectively by magnetic-activated cell sorting system. 3D cell culture was used to evaluate the ability of quercetin, MK siRNA, and the combination of both to inhibit spheroid formation, apoptosis and cell cycle arrest. Image-based cytometer, RT-qPCR, Western blotting and transwell migration assays were performed. Key findings Quercetin treatment for 24–72 h inhibited PC3 and CD44+/CD133+ stem cell proliferation in a time- and dose-dependent manner. Knockdown of endogenous MK expression significantly suppressed proliferation of CD44+/CD133+ and CD44+ cells as well as their parent cells. Co-administration of MK siRNA and quercetin reduced the cell survival, induced apoptosis and caused G1 phase cell cycle arrest more effectively than the individual therapy. Knockdown of MK significantly enhanced the inhibitory effect of quercetin on CD44+/CD133+ migration and spheroid formation. In addition, the combined therapy inhibited the phosphorylation of PI3K, AKT and ERK1/2, and reduced the protein expression of p38, ABCG2 and NF-κB. Significance Quercetin alone exhibited significant cytotoxic effects on CD44+/CD133+. MK plays an important role in the proliferation of CD44+/CD133+ and CD44+ cells in particular, and quercetin and MK-silencing therapy may be an important strategy in targeting CSCs that play a role in relapse, migration and drug resistance.

Amaç: Bu çalışmada, deneysel olarak tirotoksikoz oluşturulan sıçanlarda, tirotoksikoza bağlı olarak ovaryum yapısında, özellikle tiroid hormon reseptörleri içeren oosit, kumulus hücreleri ve gran ü losa hücrelerinde oluşabilecek histolojik değişimler ile doku rejenerasyon yeteneği ve kapasitesinin , kök hücre belirteçleri açısından imm ü nohistokimyasal olarak araştırılması amaçlan dı . Gereç ve Yöntem: Her biri 8 sıçandan oluşan 2 çalışma grubu oluşturul du. Tirotoksikoz grubu dişi sıçanlara 10 gün i ntraperitoneal 500µgr/kg/gün 3,3',5-Triiodo-L-thyronine enjekte edil di . On birinci gün tüm hayvanlara anestezi uygulan dı , biyokimyasal değerlendirmeler için 5 cc kan alındıktan sonra dekapite edilerek ovaryumları çıkarıl dı . Ovaryumlar immünohistokimyasal incelemeler için %4’ lük paraformaldehit fiksatifi içine alın dı . Rutin immünohistokimyasal takip ve boyamalar yapıl arak tüm örnekler ış ık mikroskobunda değerlendiril di . Bulgular: İmmünohistokimyasal boyamalar sonucunda c -kit ve Thy -1 ekspresyonu kontrol grubuna göre tirotoksikoz grubunda belirgin düzeyde azalmış, Nanog ekspresyonu ise tirotoksikoz grubunda artmış olarak bulundu. Tirotoksi kozlu sıçanlarda c -kit ve Thy -1’in azalması , tirotoksikozda stromal büyüme desteğinin azalmasına ve mikroçevre bozunumuna neden ol du. Sonuç: Tirotoksikozda Nanog ekspresyonunun artışı, foliküler yapıları ve oositleri korumak adına dokunun katabolik süreç (tiroid hormonlarının hemen tüm hücrelerde metabolizmayı hızlandırmasına bağlı olarak) artışına rağmen reaktif olarak mikroçevreyi koruma çabası olarak yorumlan dı ., Aim: In this study, it was aimed to investigate the possible histological changes in the ovarian structure and particularly in the oocytes, cumulus cells and granulosa cells containing thyroid hormone receptors, tissue regeneration capability and capacity in terms of stem cell markers immunohistochemically by experimentally-induced thyrotoxicosis in rats. Materials and Methods: Two study groups were formed (eight rats for each group); 500µgr/kg/day 3,3',5-Triiodo-L- thyronine was injected intraperitoneally to female rats in thyrotoxicosis group during ten days. In 11 th day, all animals were anesthesized, 5 cc blood was drawn for biochemical evaluation and then all were decapitated in order to remove ovaries. Ovaries were placed in 4% paraformaldehyde fixative for immunohistochemical investigation. Immunohystochemical processing and staining were performed routinely and all samples were evaluated by light microscopy.Results: As a result of immunohistochemical staining, c-kit and Thy-1 expressions were found to be significantly decreased in the thyrotoxicosis group compared with the control group, whereas Nanog expression was increased in the thyrotoxicosis group. Decrease in c-kit and Thy-1 expression in the thyrotoxicosis group cause d a reduction in the stimulatory effects of the growth factors on oocytes, cumulus and granulosa cells. Conclusion: Decreased levels of c-kit and Thy-1 in rat model of thyrotoxicosis lead to a decrease in stromal growth support and deterioration of microenvironment. Elevated levels of Nanog expression in thyrotoxicosis was interpreted as an effort to protect the microenvironment for the sake of follicles and oocytes despite the increase in catabolic processes (as a result of increased metabolism in almost all cells due to thyroid hormones).

Objective: Statins mayexert effects that are independent of LDL-cholesterol lowering on manycomponents of atherosclerosis including endothelial function, calledpleiotropic effects. In this study we aimed to investigate effects offluvastatin on intimal thickening in normocholesterolemic rabbits.Materials &Method: New Zeelandrabbits were treated with fluvastatin (8 mg/kg/day, p.o.) and placebo for 3weeks. On the 8th day, a non-occlusive silicon collar was positioned around theleft carotid artery for 14 days and then both carotid arteries were isolated.Intimal, medial, lumen and external elastic lamina cross sectional areas weremeasured by computer based morphometric analysis and index (intima/media),residual luminal ratio (RLR) values were calculated.Results: Collarcaused significant increase in intimal area, index values and decrease in RLRvalues. Fluvastatin treatment significantly inhibit collar-induced intimalthickening.Conclusion: Thesefindings indicate that fluvastatin may have beneficial effects on the earlystages of normocholesterolemic atherosclerosis., Amaç: StatinlerinLDL-kolesterol düşürücü etkilerinden bağımsız endotel fonksiyonu gibi aterosklerozunpek çok bileşeni üzerinde de etkileri mevcuttur. Bu etkileri pleiotropiketkileri olarak da bilinmektedir. Bu çalışmada fluvastatinin normokolesterolemiktavşanlarda intimal kalınlaşma üzerine etkisinin araştırılması planlanmıştır.Gereç ve Yöntem: Çalışmada Yeni Zelanda tavşanlarına 3 hafta boyuncafluvastatin (8 mg/kg/gün oral) ve plasebo tedavi uygulandı. Tedavinin sekizincigününde okluzif olmayan silikon bir yaka sol karotid arterinin çevresineyerleştirilerek on dört gün daha tedaviye devam edildi. Deney sonunda sağ vesol karotid arterler çıkarıldı. Çıkarılan karotid arterlerden alınan kesitlerdemorfometrik ölçümler gerçekleştirildi. İntima, medya, lümen ve eksternalelastik lamina alanları ölçülerek indeks, rezidüel lümen oranı değerlerihesaplandı.Bulgular: Yaka intimalalan ve indeks değerinde belirgin bir artışa ve RLO değerinde azalmaya nedenolurken fluvastatin yakanın neden olduğu intimal kalınlaşmayı inhibe etti.Sonuç: Bulgularımızfluvastatinin kolesterol düşürücü etkisinden bağımsız aterosklerozun erkendöneminde de yararlı etkileri olabileceğini göstermektedir.

Amaç: Zoledronik asit (ZA) uygulanmış meme kanser kök hücrelerinde (KKH), kanser kök hücresi ve otofaji ile ilişkili gen ekspresyonlarındaki farklılıkları incelemek. Gereç ve Yöntem: CD44yüksek/CD24yüksek yüzey belirteçleri pozitif olan MCF-7 meme kanseri hücre hattından KKH’ler akış sitometri metodu ile izole edilmiştir. ZA tedavisi uygulanmış ve uygulanmamış olan KKH gruplarının toplam RNA izolasyonu için cDNA sentezlenmiştir. Son olarak 41 adet otofaji için başlıca ilişkili olabilecek genlerin ekspresyon seviyeleri qRT-PCR analizi ile incelenmiştir. Bulgular: ZA tedavisi uygulanan grup tedavi uygulanmayan grup ile karşılaştırıldığında TGM2, TNF, HSPA8 genlerinin azalmış regüle olduğu ve HSP90AA1 geninin artmış regüle olduğu görülmüştür. Sonuç: Otofaji ile ilişkili genlerin ekspresyon profillerinde ZA tedavisi uygulanan ve uygulanmayan gruplar arasında anlamlı farklar bulunmuştur. Bu bulgular, KKH’lerine karşı hedef olabilecek daha etkili tedavi ajanları geliştirilebilmesi için önemli katkılar sağlayabilir., Aim: In this study we examined differences in the expression of genes associated with autophagy and CSCs for the treatment ZA applied via taking breast cancer stem cells as models. Materials and Methods: CSCs with CD44high/CD24high surface markers have been isolated from the MCF-7 breast cancer cell line with flow cytometry methods. cDNA was synthesized for ZA treatment applied and not applied CSC group with total RNA isolation. Then, 41 essential autophagy-related gene expression levels were analyzed by qRTPCR array analysis. Results: When ZA treatment administrated group compared with non-treatment group, downregulation of TGM2, TNF, HSPA8 and upregulation of HSP90AA1 was detected. Conclusion: There are differences of expression profiles of genes related to autophagy between ZA treated and non-ZA tread group. These findings may provide important information towards us to target CSCs to develop more effective treatments.

WOS: 000431261100036, PubMed ID: 29714468, Background/aim: In this study, the effects of resveratrol as a natural polyphenol compound, gemcitabine as an antimetabolite that has nucleoside structure analogous to deoxycytidine, and para-aminophenol-derived paracetamol were investigated with single and combined applications in monolayers of the MDAH-2774 human ovarian cancer cell line. Materials and methods: Drugs were evaluated in cell culture with respect to cell proliferation, cell cytotoxicity (trypan blue dye exclusion test), synthesis phase of cell cycle, and cell structure in 24, 48, 72, and 96 h. Result: Resveratrol and gemcitabine diminished both cell proliferation and cell cycle synthesis phase indication in monolayer cell cultures (P < 0.05). All combination groups showed similar effects that were mainly more effective in respect to single usage of resveratrol and gemcitabine in monolayer cell cultures. Conclusion: The effects of gemcitabine, resveratrol, and paracetamol were investigated in monolayers of the MDAH-2774 human ovarian cancer cell line and a decrease in cell number in cell cycle synthesis phase, prevention of cell proliferation, and destruction of cell structure were observed.

WOS: 000417293400016, Flavopiridol is a synthetically produced flavonoid that potently inhibits the proliferation of human tumor cell lines. Flavopiridol exerts strong antitumor activity via several mechanisms, including the induction of cell cycle arrest and apoptosis, and the modulation of transcriptional regulation. The aim of the present study was to determine the effect of flavopiridol on a subpopulation of cluster of differentiation (CD)44(+)/CD24(-) human breast cancer MCF7 stem cells. The CD44(+)/CD24(-) cells were isolated from the MCF7 cell line by fluorescence-activated cell sorting and treated with 100, 300, 500, 750 and 1,000 nM flavopiridol for 24, 48 and 72 h. Cell viability and proliferation assays were performed to determine the inhibitory effect of flavopiridol. Gene expression profiling was analyzed using Illumina Human HT-12 v4 Expression BeadChip microarray. According to the results, the half maximal inhibitory concentration (IC50) value of flavopiridol was 500 nM in monolayer cells. Flavopiridol induced growth inhibition and cytotoxicity in breast cancer stem cells (BCSCs) at the IC50 dose. The present study revealed several differentially regulated genes between flavopiridol-treated and untreated cells. The result of the pathway analysis revealed that flavopiridol serves an important role in translation, the ribosome biogenesis pathway, oxidative phosphorylation, the electron transport chain pathway, carbon metabolism and cell cycle. A notable result from the present study is that ribosome-associated gene expression is significantly affected by flavopiridol treatment. The data of the present study indicate that flavopiridol exhibits antitumor activity against CD44(+)/CD24(-) MCF7 BCSCs through different mechanisms, mainly by inhibiting translation and the ribosome biogenesis pathway, and could be an effective chemotherapeutic molecule to target and kill BCSCs.

WOS: 000344423800008, PubMed ID: 25216351, Flavopiridol is a flavone that inhibits several cyclin-dependent kinases and exhibits potent growth-inhibitory activity, apoptosis and G(1)-phase arrest in a number of human tumor cell lines. Flavopiridol is currently undergoing investigation in human clinical trials. The present study focused on the effect of flavopiridol in cell proliferation, cell cycle progression and apoptosis in prostate cancer stem cells (CSCs). Therefore, cluster of differentiation 133 (CD133)(+high)/CD44(+high) prostate CSCs were isolated from the DU145 human prostate cancer cell line. The cells were treated with flavopiridol in a dose- and time-dependent manner to determine the inhibitory effect. Cell viability and proliferation were analyzed and the efficiency of flavopiridol was assessed using the sphere-forming assay. Flavopiridol was applied to monolayer cultures of CD133(high)/CD44(high) human prostate CSCs at the following final concentrations: 100, 300, 500 and 1000 nM. The cultures were incubated for 24, 48 and 72 h. The half maximal inhibitory concentration (IC50) value of the drug was determined as 500 nM for monolayer cells. Dead cells were analyzed prior and subsequent to exposure to increasing flavopiridol doses. Annexin-V and immunofluorescence analyses were performed for the evaluation of apoptotic pathways. According to the results, flavopiridol treatment caused significant growth inhibition at 500 and 1000 nM when compared to the control at 24 h. G(0)/G(1), analysis showed a statistically significant difference between 100 and 500 nM (P

Ovarian cancer (OC) is the most lethal cancer in all of the gynecological cancers.Despite the recent advancements in the treatment of OC, it remains the seventh most common cancer in women worldwide. Cancer stem cells (CSCs) are a unique cell population described by their ability to indefinitely self-renew, proliferate, initiate, and propagate. Evidence suggest that the interaction between cancer stem cells with their microenvironment play a critical role for cancer progression. Exosomes which are the key players of microenviroment are nano-sized (50-180nm) endosomal pathway-derived vesicles that present in almost all biological fluids.Transporting information via exosomes and exosomal miRNAs is deemed to be the crucial way of intercellular communication that is as essential as the cell-to-cell contact-dependent signaling. As a key regulator of cellular signaling, both normal, and cancer stem cells secrete exosomes to orchestrate diverse autocrine and paracrine functions which alter tumor micro-environment, growth, progression and putative immunological function. There is still not much known about how ovarian cancer stem cells (OCSCs) interact with their tumor microenvironment to promote oncogenic phenotype. Thus, in this study we investigate whether the OCSCs-derived exosomal miRNAs mirror and orchestrate that of the tumor microenvironment and thus could be used diagnostically in ovarian cancer patients.To this end,we evaluated significantly differentially expressed miRNAs between OCSCs exosomes and their cells of origin using the Affymetrix Gene Chip miRNA 4.0 microarrays. Several miRNAs were found to be significantly upregulated in both OCSCs and their exosomes. In addition we found many common up/down regulated miRNAs (miR-103a-3p, miR-20a-5p, miR-125b-5p, miR-17-5p, miR-27a-3p, miR-8075, miR-4487) with the microarray data in which tumor-associated macrophage (TAM) exosomes and their cells of origin were compared. We also found that OCSCs secrete exosomes enriched in miR-103a-3p, which has been shown to increases cancer migration, invasion and angiogenesis and induces polarization of M0 type macrophages toward immunosuppressive M2-type macrophages. In addition TAMs promote cancer stem cell-like properties by enhancing their epithelial–mesenchymal transition capacity. We are currently validating the identified miRs and investigating the role of these miRs in OC cells and CSCs proliferation, migration, invasion and tumor growth. Our findings also indicate that the interaction between TAMs and OCSCs are bidirectional. Overall, our data suggest that OCSCs and OCSCs secreted exosomes may contribute tumor microenvironment especially by regulating TAMs by transferring oncogenic miRs and using them as a progression and invasion tools within tumor microenvironment. Citation Format: Fahriye Duzagac, Gabriel Lopez-Berestein, Pinar Kanlikilicer, Cristian Rodriguez-Aguoya, Roberto Cardenas-Zuniga, Ummu Guven, Gulperi Oktem, Bulent Ozpolat. Regulatory role cancer stem cells (CSCs)-derived exosomal miRNAs in ovarian cancer growth and progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3686.

Statins are widely used besides their myopathic side effects, ranging from mild myalgia to fatal rhabdomyolysis. Resveratrol is one of the most popular over the counter products used for similar purposes with statins. The aim of this study was to elucidate the myopathic effects of atorvastatin and coadministered resveratrol in male rat skeletal muscle via morphological analyses and immunohistochemistry studies. Control group received 1.5 ml of drinking water by oral gavage and 1 ml 15% ethanol (vehicle of resveratrol) i.p. for 14 days daily; atorvastatin group was treated with 40 mg/kg atorvastatin by oral gavage and 1 ml 15% ethanol i.p. for 14 days daily. Resveratrol + atorvastatin group was treated with 40 mg/kg atorvastatin by oral gavage and 20 mg/kg i.p resveratrol for 14 days daily. Atorvastatin treatment resulted with a moderate inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) immunoreactivity in nucleus and strong immunoreactivity in fibers. Control group and resveratrol + atorvastatin group showed weak iNOS and eNOS immunoreactivity in nucleus and moderate immunoreactivity muscular fibers. Treatment with atorvastatin resulted in a significantly shortened fibrils, and resveratrol co-treatment reversed this effect. Resveratrol and atorvastatin co-treatment could be an alternative treatment to prevent the myositis adverse effects of atorvastatin on skeletal muscle. Key words: Atorvastatin, myositis, resveratrol.

Amaç: Tirozin kinaz inhibisyonunun diyabet etkisi altındaki testis dokusu üzerine göstereceği etkileri araştırmaktır. Gereç ve Yöntem: Çalışmamızda 31 adet CD1 türü erkek fare kullanıldı ve dört gruba ayrıldı: Grup 1'de (kontrol grubu) 7, Grup 2'de tirozin kinaz inhibitörü uygulanan 7, Grup 3'te diyabetik ve SF uygulanan 8, Grup 4'te diyabet + tirozin kinaz inhibitörü uygulanan 9 denek hayvanı yer aldı. Grup 1'de herhangi bir uygulama yapılmadı. Grup 2'deki farelere 3 hafta boyunca tirozin kinaz inhibitörü verildi. Diyabet oluşturulması için 0.1mol/L tek doz streptozotosinin intraperitoneal olarak verildi. 250 mg/dL ve üzeri kan glikoz seviyesi diyabetik olarak kabul edildi. Deneysel diyabet modeli oluşturulan farelere 1 hafta beklendikten sonra, Grup 3'e SF, Grup 4'e 3 hafta boyunca tirozin kinaz inhibitörü verildi. Sonunda tüm denek hayvanları anestezi altında sakrifiye edilerek histopatolojik inceleme için testis dokuları alındı. İstatistiksel analiz için tek yönlü varyans analizi (ANOVA) testi yapıldı, 0.05'ten küçük p değerleri, istatistiksel olarak anlamlı kabul edildi. Bulgular: Testis dokusu histopatolojik olarak incelendiğinde deneysel diyabete bağlı olarak seminifer tübülün germ hücre serilerinde kayıp, hücre bütünlüklerinde ise bozulma saptandı. Sonuç: Bu çalışma, diyabetin testiste germ hücre serilerinde sayısal olarak azalmaya ve hücre adezyon mekanizmasında bozulmaya yol açtığını göstermektedir. Tirozin kinaz inhibitörü uygulamasının, bu hasarlanmada tamir edici etkisinin olduğu düşünülmektedir. Bu hasarın tedavisinin derecesi, uygulanan tirozin kinaz inhibitörünün dozu ve süresine bağlı olarak farklılık gösterebilmektedir. Ancak, klinik diyabet uygulamalarında tirozin kinaz inhibitörü kullanılabilmesi için bu konuda moleküler çalışma sayılarının artışına ihtiyaç vardır., Aim: The aim of the study is to investigate the effects of tyrosine kinase inhibition on testicular tissue under the effects of diabetes mellitus. Materials and Methods:. Thirty one CD-1 male mice were divided into four groups: Group 1 (control group) consited 7, Group 2 (tyrosine kinase inhibitor given group) 7, Group 3 (diabetic group treated with SF) 8 and Group 4 (diabetic group treated with tyrosine kinase inhibitor) 9 experimental mice. No application was made to the animals in Group 1. In Group 2, mice were given tyrosine kinase inhibitor for 3 weeks. Diabetes was induced by intraperitoneal injection of single dose of streptozotocin 0.1mol/L. Mice with 250 mg/dL and higher blood glucose levels were accepted as diabetic. After waiting for 1 week, experimental diabetic mice in Group 3 were treated with SF and those in Group 4 were treated with tyrosine kinase inhibitor for 3 weeks. Then, all the experimental animals were sacrified and the testicular tissue was removed for histopathological examination.The one-way analysis of variance (ANOVA) is used for statistical analysis, a p value less than 0.05 was considered statistically significant. Results: Histopathologic examination of the testes revealed loss of germ cells series in the seminiferous tubules and deterioration in cellular integrity depending on experimental diabetes. Conclusion: By the study, it was demonstrated that diabetes lead to a decrease in the number of germ cells in testes and deterioration in cell adhesion mechanism. The administration of tyrosine kinase inhibitor could be considered to have a regenerative effect in this damage. The degree of treatment of this injury varies depending to the dose and time of the tyrosine kinase inhibitors. But, for the use of tyrosine kinase inhibitors in clinical practice of diabetes mellitus, more molecular studies are needed.

WOS: 000416401500005, PubMed ID: 28994095, Introduction. Sunitinib is an oral inhibitor of vascular endothelial growth factor that is used to treat a variety of cancer. There are limited data regarding the effect of sunitinib on diabetes. In the liver, Notch signaling plays an important role in liver tissue development and homeostasis and its dysfunction is associated with liver pathologies. The aim of the present study is to investigate the effects of sunitinib on streptozotocin (STZ)-induced diabetic liver in mice models. Material and methods. An experimental diabetes mellitus (DM) model was created in 28 male CD-1 mice. Twenty-eight male CD-1 mice divided in four groups (n = 7 each) were used; control mice (C), control mice treated with sunitinib (C + S), diabetic mice (DM), and diabetic mice treated with sunitinib (DM + S) for four weeks. The histopathological changes in the liver were examined by histochemistry and immunohistochemistry. Immunoreactivity of Notch1, Jagged1, DLL-1 and VEGF were evaluated in control and diabetic mice after sunitinib treatment. Results. The significant morphological changes in the liver were mostly seen in hepatocytes that were hypertrophied in the DM mice, with an increased amount of eosinophilic granules; moreover, some hepatocytes contained empty vacuole-like structures. The livers of the DM mice revealed increased deposition of collagen fibers. After sunitinib treatment the hepatocytes and hepatic lobules had almost similar morphology to control mice. The immunoreactivities of Notch1, Jagged1, DLL-1 and VEGF in hepatocytes were significantly lower in the DM group when compared with the C, DM + S and C + S group treated with sunitinib. Conclusions. These results suggest that sunitinib effectively protects the liver from diabetes-induced damage through the inhibition of the Notch pathway.

Flavopiridol is a synthetically produced flavonoid that potently inhibits the proliferation of human tumor cell lines. Flavopiridol exerts strong antitumor activity via several mechanisms, including the induction of cell cycle arrest and apoptosis, and the modulation of transcriptional regulation. The aim of the present study was to determine the effect of flavopiridol on a subpopulation of cluster of differentiation (CD)44+/CD24− human breast cancer MCF7 stem cells. The CD44+/CD24− cells were isolated from the MCF7 cell line by fluorescence-activated cell sorting and treated with 100, 300, 500, 750 and 1,000 nM flavopiridol for 24, 48 and 72 h. Cell viability and proliferation assays were performed to determine the inhibitory effect of flavopiridol. Gene expression profiling was analyzed using Illumina Human HT-12 v4 Expression BeadChip microarray. According to the results, the half maximal inhibitory concentration (IC50) value of flavopiridol was 500 nM in monolayer cells. Flavopiridol induced growth inhibition and cytotoxicity in breast cancer stem cells (BCSCs) at the IC50 dose. The present study revealed several differentially regulated genes between flavopiridol-treated and untreated cells. The result of the pathway analysis revealed that flavopiridol serves an important role in translation, the ribosome biogenesis pathway, oxidative phosphorylation, the electron transport chain pathway, carbon metabolism and cell cycle. A notable result from the present study is that ribosome-associated gene expression is significantly affected by flavopiridol treatment. The data of the present study indicate that flavopiridol exhibits antitumor activity against CD44+/CD24− MCF7 BCSCs through different mechanisms, mainly by inhibiting translation and the ribosome biogenesis pathway, and could be an effective chemotherapeutic molecule to target and kill BCSCs.

Similarities and differences in the cell cycle components, apoptosis and cytoskeleton-related molecules among mouse skin fibroblast cells (MSFs), mouse squamous cell lung carcinomas (SqCLCs) and mouse embryonic stem cells (mESCs) are important determinants of the behaviour and differentiation capacity of these cells. To reveal apoptotic pathways and to examine the distribution and the role of cell cycle-cell skeleton comparatively would necessitate tumour biology and stem cell biology to be assessed together in terms of oncogenesis and embryogenesis. The primary objectives of this study are to investigate the effects of flavopiridol, a cell cycle inhibitor, and geldanamycin, a heat shock protein inhibitor on mouse somatic, tumour and embryonic stem cells, by specifically focusing on alterations in cytoskeletal proteins, cell polarity and motility as well as cell cycle regulators. To meet these objectives, expression of several genes, cell cycle analysis and immunofluorescence staining of intracellular cytoskeletal molecules were performed in untreated and flavopiridol- or geldanamycin-treated cell lines. Cytotoxicity assays showed that SqCLCs are more sensitive to flavopiridol than MSFs and mESCs. Keratin-9 and keratin-2 expressions increased dramatically whereas cell cycle regulatory genes decreased significantly in the flavopiridol-treated MSFs. Flavopiridol-treated SqCLCs displayed a slight increase in several cell cytoskeleton regulatory genes as well as cell cycle regulatory genes. However, gene expression profiles of mESCs were not affected after flavopiridol treatment except the Cdc2a. Cytotoxic concentrations of geldanamycin were close to each other for all cell lines. Cdkn1a was the most increased gene in the geldanamycin-treated MSFs. However, expression levels of cell cytoskeleton-associated genes were increased dramatically in the geldanamycin-treated SqCLCs. Our results revealing differences in molecular mechanisms between embryogenesis and carcinogenesis may prove crucial in developing novel therapeutics that specifically target cancer cells.

Objective To describe the effects of epididymal and vasal ligation, in an experimental rat model, by using connexin 43 and occludin immunohistochemistry as well as transmission electron microscopy. Design Comparative and controlled experimental research study. Setting University animal research and histology laboratories in Turkey. Animal(s) Wistar male rats in experimental and control groups. Intervention(s) The control group underwent sham operation (n = 7). The first experimental group (n = 7) underwent unilateral epididymal ligation, whereas the second experimental group (n = 7) underwent unilateral vasal ligation to induce experimental epididymal and vasal obstruction models, respectively. All animals were then killed at 90 days. Main Outcome Measure(s) Immunohistochemical expression of connexin 43 and occludin for testicular tissues was determined after epididymal and vasal obstruction models. Ultrastructural morphological changes were examined by electron microscopy. Result(s) Results of the semiquantitative analysis revealed that expressions of both occludin and connexin 43 in the rat testis were decreased in the experimental groups compared with in the sham-operated group. However, changes after vasal ligation were more prominent. Ultrastructural examination confirmed decreased intercellular communication as well as increased cellular degeneration among the ipsilateral and contralateral testicular tissues. Conclusion(s) Immunohistochemical expression of occludin and connexin 43 were decreased in the testis after vasal and epididymal ligation when compared with the sham-operated group. Ultrastructural changes indicating cell degeneration were more prominent after vasal ligation.

WOS: 000378324000020, PubMed ID: 26981753, The aim of this study was to evaluate whether medium modification improves the odontogenic differentiation of human dental pulp stem cells (DPSC) in vitro and in vivo. DPSC isolated from human impacted third molar teeth were analysed for clusters of differentiation with flow cytometry. Odontogenic differentiation was stimulated by medium modification with the addition of bone morphogenetic protein 2 (BMP2). The expression of dentin sialophosphoprotein, dentin matrix protein 1, enamelysin/matrix metalloproteinase 20 and the phosphate-regulating gene with homologies to endopeptidases on the X chromosome of the cells were analysed with RT-PCR at 7, 14 and 21 days. Then, DPSC were transplanted on the back of immunocompromised mice via a hydroxyapatite tricalcium phosphate scaffold, and the structure of the formed tissue was investigated. The cells were identified as mesenchymal stem cells with a 98.3% CD73 and CD90 double-positive cell rate. The increase in mineralization capacity and expression of human enamel-dentin specific transcripts proportional to the culture period were determined after differentiation. Six weeks after transplantation, an osteo-dentin matrix was formed in the group in which odontogenic differentiation was stimulated, and the odontogenic characteristics of the matrix were confirmed by histological examination and RT-PCR analysis. Odontogenic differentiation of the isolated and characterized human DPSC was improved with medium modification by the addition of BMP2 in vitro and in vivo. The defined medium and applied technique have a potential use for forming reparative dentin in the future, but the effects of the method should be investigated in long-term studies., Ege University's Scientific Research FoundationEge University [2009-Dis-036]; Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK), This study was supported by Ege University's Scientific Research Foundation (2009-Dis-036). We also acknowledge The Scientific and Technological Research Council of Turkey (TUBITAK) for its PhD scholarship support.

PubMed ID: 27787370, Background: Flavopiridol a semisynthetic flavone that inhibits cyclin-dependent kinases (CDKs) and has growth-inhibitory activity and induces a blockade of cell-cycle progression at G1-phase and apoptosis in numerous human tumor cell lines and is currently under investigation in phase II clinical trials. Cancer stem cells (CSCs) are comprised of subpopulation of cells in tumors that have been proposed to be responsible for recurrence and resistance to chemotherapy. The aim of the present study was to investigate the effects of flavopiridol in cancer stem cell cytoskeleton, cell adhesion, and epithelial to mesenchymal transition in CSCs. Methods:The cells were treated with flavopiridol to determine the inhibitory effect. Cell viability and proliferation were determined by using the WST-1 assay. Caspase activity and immunofluorescence analyses were performed for the evaluation of apoptosis, cell cytoskeleton, and epithelial-mesenchymal transition (EMT) markers. The effects of flavopiridol on the cell cycle were also evaluated. Flow cytometric analysis was used to detect the percentages of CSCs subpopulation. We analyzed the gene expression patterns to predict cell cycle and cell cytoskeleton in CSCs by RT-PCR. Results: Flavopiridol-induced cytotoxicity and apoptosis at the IC50 dose, resulting in a significant increase expression of caspases activity. Cell cycle analyses revealed that flavopiridol induces G1 phase cell cycle arrest. Flavopiridol significantly decreased the mRNA expressions of the genes that regulate the cell cytoskeleton and cell cycle components and cell motility in CSCs. Conclusion: Our results suggest that Flavopiridol has activity against lung CSCs and may be effective chemotherapeutic molecule for lung cancer treatment. © Copyright 2016 the Author(s). Published by Wolters Kluwer Health, Inc. All rights reserved.

WOS: 000404458100052, PubMed ID: 27901202, This study was designed to determine the in vivo performance of three different materials as scaffolds for dental pulp stem cells (DPSC) undergoing induced odontogenic differentiation. An odontogenic medium modified by the addition of recombinant human bone morphogenetic protein 2 was used in the experimental groups to induce differentiation. Mesenchymal stem cell medium was used in the control groups. DPSC were transplanted onto the backs of mice via three scaffolds: copolymer of L-lactide and DL-lactide (PLDL), copolymer of DL-lactide (PDL) and hydroxyapatite tricalcium phosphate (HA/TCP). The expression levels of dentin sialo-phosphoprotein (DSPP), dentin matrix protein-1 (DMP1), enamelysin/matrix metalloproteinase 20 (MMP20) and phosphate-regulating gene with homologies to endopeptidases on X chromosome (PHEX) were analysed using RT-PCR. The expressions in the experimental groups were compared to those in the control groups. The transcript expressions at 6 and 12 weeks were significantly different for all scaffolds (p < 0.05), except for the expression of DSPP in the PLDL group with regard to the time variable. Although there was a decrease in the expression of enamelysin/MMP20 in PLDL and HA/TCP at 12 weeks, all other expressions increased and reached their highest level at 12 weeks. The highest DSPP expression was in the PDL group (p < 0.05). The highest expression of DMP1 was detected in the HA/TCP group (p < 0.05). The highest expression of PHEX was in the PLDL group (p < 0.05). Consequently, PLDL and PDL seemed to be promising scaffold candidates for odontogenic regeneration at least as HA-TCP, when they were applied with the DPSC induced for odontogenic differentiation., Ege University's Scientific Research FoundationEge University [2010-Dis-006], This study was supported by Ege University's Scientific Research Foundation (2010-Dis-006). The authors sincerely thank Gul KAYRAK for her help in language editing.

JAK/STAT is an evolutionarily conserved pathway and very important for second messenger system. This pathway is important in malignant transformation and accumulated evidence indicates that this pathway is involved in tumorigenesis and progression of several cancers. It was possible to assume that activation of JAK/STAT pathway is associated with increase in the expressions of ICAM/1 and VCAM-1. In this study we hypothesized that when cells were maintained as spheroids or monolayers, the structure of cancer stem cells (CSCs) could show differentiation when compared with non-CSCs.DU-145 human prostate cancer cells were cultured using the Ege University molecular embryology laboratory medium supplemented wıth 10% fetal bovine serum. Clusters of differentiation 133 (CD133)(+high)/CD44(+high) prostate CSCs were isolated from the DU145 cell line by using BD FACSAria. CD133//CD44+ CSCs were cultured until confluent with 3% noble agar. The expression of these proteins in CSCs and non-CSCs was analyzed by immunohistochemistry.Different expression profiles were observed in the conventional two-dimensional (2D) and three-dimensional (3D) experimental model system when CSCs and non-CSCs were compared. Human prostate CSCs exhibited intense ICAM-1 and VCAM-1 immunoreaction when compared with non-CSCs. These findings were supported by the fact that VCAM-1 on the surface of cancer cells binds to its counterreceptor, the α4β1 integrin (also known as very-late antigen, VLA-4), on metastasis-associated macrophages, triggering VCAM-1-mediated activation of the phosphoinositide 3-kinase growth and survival pathway in cancer cells.The results of this study showed that changes in JAK/STAT pathway are related with adhesion molecules and could affect cancer progression.

Collagen scar formation at the cut end of a nerve, an important problem in clinical practice for neurosurgeons in peripheral nerve surgery, obstructs sprouting of axons into appropriate distal fascicles, and thereby limits nerve regeneration. Researchers attempt to control collagen accumulation in the formation of neuroma by various physical and chemical methods, but these have yielded only limited functional success. This is the first experimental study investigating the effects of melatonin (MLT) on nerve repair and neuronal regeneration in rat sciatic nerve suture repair.The hypothesis that exogenous MLT administration may inhibit the formation of neuroma in peripheral nerve surgery was investigated in rat sciatic nerve model. In this study, a total of 80 rats were used for control groups (Groups Ia, Ib, IIa, and IId), MLT group (Group Ic), surgical pinealectomy (Px) groups (Groups IIb and IIc), and group of MLT treatment following Px procedure (Group IIe). All animals underwent a surgical intervention consisting of bilateral sciatic nerve section and primary suture repair. At 8 weeks after repair, the animals were killed following completion of recording of nerve action potentials (NAPs). Then, unilateral sciatic nerve specimens including the suture repair region were carefully removed and the excised segments were processed for electron microscopy examination. Afterwards, contralateral sciatic nerve specimens from two animals from each group were removed and stained for immunohistochemical analysis.Results of morphometric analysis revealed that Px procedure caused an elevation of collagen content of the sciatic nerve and macroscopic neuroma formation, and that there was a statistically significant reduction in collagen content of the same region in pinealectomized animals treated with MLT (p0.001). Accordingly, electrophysiological findings demonstrated that the stimulus intensities required to excite a NAP response were increased in surgical Px group, but the presence of a reduced threshold response was found in the group treated with MLT following Px procedure (p0.01). Immunohistochemical staining for Type I collagen and Type III collagen was markedly more intense in the epineurium of animals after Px. Virtually no or only weak staining was observed in animals in control groups and the MLT treatment group. Results of immunohistochemical analysis revealed that surgical Px procedure caused a strong immunoreactivity for Type I collagen and Type III collagen in all connective tissue planes of the nerve, especially in the epineurium, and there was a statistically significant reduction in immunoreactivity of the repair region in animals receiving MLT treatment after Px procedure (p0.001).This study demonstrates that exogenous MLT administration significantly inhibits collagen accumulation in the formation of neuroma in the suture repair site and thereby improves nerve regeneration. From a clinical standpoint, the positive effect of MLT administration on neuroma formation and nerve regeneration seems a particularly attractive treatment option. Therefore, we believe that nerve repair with addition of MLT may be a worthwhile option in addition to other treatment modalities in case of MLT deficiency, such as aging. However, further experimental and clinical studies using functional analysis warranted to confirm this result in future.

PubMed ID: 26485709, Trabectedin (Yondelis, ET-743) is a marine-derived tetrahydroisoquinoline alkaloid. It is originally derived from the Caribbean marine tunicate Ecteinascidia turbinata and currently produced synthetically. Trabectedin is active against a variety of tumor cell lines growing in culture. The present study focused on the effect of trabectedin in cell proliferation, cell cycle progression, apoptosis and spheroid formation in prostate cancer stem cells (CSCs). Cluster of differentiation (CD) 133+high/CD44+high prostate CSCs were isolated from the DU145 and PC-3 human prostate cancer cell line through flow cytometry. We studied the growth-inhibitory effects of trabectedin and its molecular mechanisms on human prostate CSCs and non-CSCs. DU-145 and PC-3 CSCs were treated with 0.1, 1, 10 and 100 nM trabectedin for 24, 48 and 72 h and the growth inhibition rates were examined using the sphere-forming assay. Annexin-V assay and immunofluorescence analyses were performed for the detection of the cell death. Concentration-dependent effects of trabectedin on the cell cycle were also evaluated. The cells were exposed to the different doses of trabectedin for 24, 48 and 72 h to evaluate the effect of trabectedin on the number and diameter of spheroids. According to the results, trabectedin induced cytotoxicity and apoptosis at the IC50 dose, resulting in a significant increase expression of caspase-3, caspase-8, caspase-9, p53 and decrease expression of bcl-2 in dose-dependent manner. Cell cycle analyses revealed that trabectedin induces dose-dependent G2/M-phase cell cycle arrest, particularly at high-dose treatments. Three dimensional culture studies showed that trabectedin reduced the number and diameter of spheroids of DU145 and PC3 CSCs. Furthermore, we have found that trabectedin disrupted cell-cell interactions via E-cadherin in prostasphere of DU-145 and PC-3 CSCs. Our results showed that trabectedin inhibits cellular proliferation and accelerates apoptotic events in prostate CSCs; and may be a potential effective therapeutic agent against prostate cancer. © Copyright: 2015 Acikgoz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

WOS: 000337472500001, PubMed ID: 24971353, Background. The aim of this study is to determine the effects of zinc and/or progesterone via the expression of alpha v beta 5 integrins and Vitronectins and embryonic stem cell markers during the peri-implantation period. Methods. Four experimental groups were organized. All subjects were mated with males of the same strain to induce pregnancy; after 5 days, zinc and/or progesterone were administered. Blood levels of zinc and progesterone were determined on the sixth day and endometrial tissues were obtained in order to evaluate the immunohistochemical expression of integrins and embryonic stem cell markers. Results. The alpha v beta 5 integrin and vitronectin expression increased in the zinc group compared with the control group and no difference in the progesterone group and zinc + progesterone group. Expression of Klf-4, Sox-2, and c-Myc was found to be increased in the zinc group compared to controls, while no difference was determined between the progesterone, zinc + progesterone, and control groups. Distinctively, expression of the embryonic stem cell marker Oct-4 was increased in all of the experimental groups. Conclusions. Expression of alpha v beta 5 integrin, vitronectin, and embryonic stem cell markers might be increased by the administration of zinc. Our results suggest that zinc could be useful in the induction of implantation rates., Ege University Faculty of Medicine-Research Funds, Izmir, TurkeyEge University [2010-TIP-059], The authors would like to thank the technical lab staff from the Department of Histology & Embryology in Ege University, Faculty of Medicine, Izmir, Turkey, for their help. This study is the doctorate thesis of the first author and has been funded by Ege University Faculty of Medicine-Research Funds Project no. 2010-TIP-059, Izmir, Turkey. No commercial support was obtained for this study.

Established cancer cell lines contain cancer stem cells (CSCs) which can propagate to form three dimensional (3D) tumor spheroids in vitro. Aberrant activation of WNT signaling is strongly implicated in the progression of cancer and controls CSCs properties. In this study we hypothesized that when cells were maintained as spheroids, the structure of CSCs could show differentiation between CSCs and non- CSCs.CD133+/CD44+ cancer-initiating cells were isolated from DU-145 human prostate cancer cell line monolayer cultures, propagated as tumor spheroids and compared with the remaining heterogeneous cancer cells bulk population. The expression levels of WNT1, FZD1, ADAR, APC, AXIN, BTRC, FRAT1 and PPARD genes were measured by polymerase chain reaction (PCR) array assay and the protein expression levels of WNT1, FZD and AXIN by immunohistochemistry.The expression levels of WNT pathway-related molecules were found to increase in both CSCs and non- CSCs when CSCs were maintained as spheroids. However, different expression profiles were observed when CSCs and non-CSCs were compared. In spheroids, the expression levels of FZD1, APC, ADAR, WNT1, PPARD genes in CSCs decreased when compared to non-CSCs. Interestingly, when CSCs from spheroids were compared with CSCs from monolayers the most significant decrease was observed in FZD1 and increase in APC genes.It is possible to assume that intracellular signaling of WNT-related molecules in the nucleus and/or cytoplasm might play an important role but it is independent from increased ligand expression and this expression strongly differentiate CSCs and non-CSCs population. This unexpected expression could be important for CSCs behavior and targeting this pathway could have therapeutic implications in cancer.

WOS: 000336555200071, Cancer stem cells (CSC) isolated from multiple tumor types differentiate in vivo and in vitro when cultured in serum; however, the factors responsible for their differentiation have not yet been identified. The first aim of the present study was to identify CD133(high)/CD44(high) DU145 prostate CSCs and compare their profiles with non-CSCs as bulk counterparts of the population. Subsequently, the two populations continued to be three-dimensional multicellular spheroids. Differentiation was then investigated with stem cell-related genomic characteristics. Polymerase chain reaction array analyses of cell cycle regulation, embryonic and mesenchymal cell lineage-related markers, and telomerase reverse transcriptase (TERT) and Notch signaling were performed. Immunohistochemistry of CD117, Notch1, Jagged1, Delta1, Sox2, c-Myc, Oct4, KLF4, CD90 and SSEA1 were determined in CSC and non-CSC monolayer and spheroid subcultures. Significant gene alterations were observed in the CD133(high)/CD44(high) population when cultured as a monolayer and continued as spheroid. In this group, marked gene upregulation was determined in collagen type 9 a1, Islet1 and cyclin D2. Jagged1, Delta-like 3 and Notch1 were respectively upregulated genes in the Notch signaling pathway. According to immunoreactivity, the staining density of Jagged1, Sox2, Oct4 and Klf-4 increased significantly in CSC spheroids. Isolated CSCs alter their cellular characterization over the course of time and exhibit a differentiation profile while maintaining their former surface antigens at a level of transcription or translation. The current study suggested that this differentiation process may be a mechanism responsible for the malignant process and tumor growth., Ege University Scientific Research Project FundEge University [Ege-BAP 2008 Tip-019], The authors would like to thank Professor Benjamin Bonavida for his editorial expertise; Assistant Professor Ogun Sercan for statistical analysis; Ummu Guven and Turker Cavusoglu for their support in Cell Culture Laboratory maintenance and inter-laboratory correlations; and Tayyibe Yeni and Merve Kursuner for flow cytometry and cell-sorting experiments in AREL. The current study was funded by a grant from the Ege University Scientific Research Project Fund (no. Ege-BAP 2008 Tip-019).

Diabetes mellitus (DM) is a multisystem disorder. Type 1 DM can be experimentally induced in rats with streptozotocin (STZ). Diabetic conditions result in testicular oxidative stress and suppressed male reproductive activity as well as decreases in both testicular organ weights and subject weights.The purpose of this study was to investigate immunohistochemical differences in testicular tissue due to STZ induced diabetes regarding pluripotency via transcription factors like Klf4, Sox2, c-Myc and Oct4, and to determine weight changes in both the subjects and the testes during the experiment.Diabetes was induced in male adult rats for this study. A healthy control group and a diabetic group were observed for one month. Blood glucose levels over 250 mg/dL were considered diabetic.On days 0, 3, 15 and 30, the subjects' weights and testicular organ weights were determined and analyzed. The results revealed statistically significant decreases (p0.05 and p0.001, respectively). Semiquantitative immunohistochemical analyses of Klf4, Sox2, c-Myc and Oct4 were studied in testes paraffin sections via light microscopy. Decreased immunoreactivity of Klf4 was observed in the diabetic group in comparison to the controls. Spermatogonial cells and Sertoli cells showed increased immunostaining for Sox2 and c-Myc, while decreased immunoreactivity of Oct4 was noted for both spermatogenic and Sertoli cells compared to the control group.This study clearly demonstrated that Klf4, Sox2 and Oct4 immunopositive cells in adult male rat testes manifested sustainable pluripotency and that diabetes has dramatically detrimental effects on this trait.