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Cancer stem cells (CSCs) are a unique population that has been linked to drug resistance and metastasis and recurrence of prostate cancer. The sonic hedgehog (SHH) signal regulates stem cells in normal prostate epithelium by affecting cell behavior, survival, proliferation, and maintenance. Aberrant SHH pathway activation leads to an unsuitable expansion of stem cell lineages in the prostate epithelium and the transformation of prostate CSCs (PCSCs). Zoledronic acid (ZOL), one of the third-generation bisphosphonates, effectively prevented bone metastasis and treated advanced prostate cancer despite androgen deprivation therapy. Despite strong evidence for the involvement of the SHH in human PCSCs survival and drug resistance, the roles of SHH in the PCSCs-related resistance to ZOL remain to be fully elucidated. The present study aimed to investigate the role of the SHH pathway in ZOL resistance of PCSCs in 2D and three 3D cell culture conditions. For this purpose, we isolated CD133(high)/ CD44(high) PCSCs using a flow cytometer. Following ZOL treatment, mRNA and protein expressions of the components of the SHH signaling pathway in PCSCs and non-CSCs were analyzed using qRT-PCR and Immunofluorescence staining, respectively. Our finding suggested that SHH signaling may be activated by different mechanisms that lead to avoidance of the inhibition effect of ZOL. Thereby, SHH pathways may be associated with the resistance to ZOL developed by prostate CSCs. Inhibition of CSCs-related SHH signaling along with ZOL treatment should be considered to achieve improvement in survival or delayed treatment failure and prevention of the CSCs-related drug resistance., Ege University Research Foundation [2016-TIP-079], This work was supported by Ege University Research Foundation (Project no: 2016-TIP-079).

High recurrence and metastatic behavior patterns are the most important reasons for the failure of treatment strategies in patients with colon cancer. Cancer stem cells (CSCs), which are considered root of cancer, are thought to be associated with therapy resistance, relapse, and metastasis, and, therefore, targeting CSCs rather than the bulk population may be an effective approach. In cancer studies, there is an increasing interest in close friendship between epithelial-mesenchymal transition (EMT) and CSCs. Triptolide (TPL) isolated from Chinese herb Tripterygium wilfordii has important effects on the prevention of migration and metastasis as well as cytotoxic effect against cancer cells. The potential lethal efficacy of TPL on CSCs that is highly resistant to the drug is an unsolved mystery. Fundamentally, the present study basically aims to find answers to two questions: (a) is it possible to target colon CSCs with TPL? and (b) what are the mechanisms underlying TPL's potential to eliminate CSCs? Cytotoxic effects of TPL on CSCs were evaluated by WST-1 and Muse count and viability assays. Apoptosis assay and cell-cycle analysis were performed to investigate the inhibitory effect of TPL. Moreover, the effects of TPL on spheroid formation capacity, migration, and EMT processes, which are associated with CSC phenotype, were also investigated. The results revealed that TPL triggered cell death and apoptosis and altered cell cycle distribution. Moreover, TPL significantly reduced the snail slug and twist expressions associated with EMT. TPL has been shown to be effective in colon CSCs by in vitro experiments, and it might be a highly effective agent against colon cancer has been implicated in need of supporting in vivo and clinical studies.

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

Cancer stem cells (CSCs), which represent the root cause of resistance to conventional treatments, recurrence, and metastasis, constitute the critical point of failure in cancer treatments. Targeting CSCs with dendritic cell (DC)-based vaccines have been an effective strategy, but sialic acids on the surface of DCs limit the interaction with loaded antigens. We hypothesized that removal of sialic acid moieties on immature DCs (iDCs) could significantly affect DC-CSC-antigen loading, thereby leading to DC maturation and improving immune recognition and activity. The lysate of CD44+/CD24-/low breast CSCs (BCSCs) was pulsed with sialidase-treated DCs to obtain mature dendritic cells (mDCs). The roles of cytoskeletal elements in antigen uptake and dendritic cell maturation were determined by immunofluorescence staining, flow cytometry, and cytokine measurement, respectively. To test the efficacy of the vaccine in vivo, CSCs tumor-bearing mice were immunized with iDC or mDC. Pulsing DCs with antigen increased the expression levels of actin, gelsolin, talin, WASp, and Arp2, especially in podosome-like regions. Compared with iDCs, mDCs expressed high levels of CD40, CD80, CD86 costimulatory molecules and increased IL-12 production. Vaccination with mDC: i) increased CD8+ and CD4 + T-cell numbers, ii) prevented tumor growth with anti-mitotic activity and apoptotic induction, iii) suppressed metastasis by decreasing Snail, Slug, and Twist expressions. This study reveals for the first time that sialic acid removal and loading with CSC antigens induces significant molecular, morphological, and functional changes in DCs and that this new DC identity may be considered for future combined immunotherapy strategies against breast tumors., Ege University Faculty of Medicine Histology and Embryology Laboratories, Partial financial support was received from the OYP (Academician training program of Turkey) and Ege University Faculty of Medicine Histology and Embryology Laboratories.

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.

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.

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).

Physiology of the mammalian body has been adapted to diurnal cycles of around 24 h, an evolutionary situation that affects a wide spectrum of biological events including sleep-to-wake transitions, feeding/fasting, body temperature, and hormonal regulations. The patterns of the diurnal cycle occur due to rhythmic oscillations that arise from the suprachiasmatic nucleus of hypothalamus, which also can be defined as the pacemaker of the system. The clock can be defined as a molecular machinery driven by the core clock genes that encode clock proteins in a rhythmic oscillatory fashion maintained by the light/dark cycles of the environment. Although the well-established knowledge refers to the function of the circadian rhythm as maintenance of the normal physiology, growing evidence shows that disruptions in the system usually caused by genetic and/or epigenetic misregulations may have a direct effect to lead major pathological conditions, such as carcinogenesis. This review outlines the main molecular aspects of circadian physiology, and reveals the reasons for and results of the circadian disruptions at different levels. In spite of the fact that more proof is needed for a direct correlation between circadian disruptions and oncogenesis and other pathological events, data obtained from current research supports the role of circadian rhythms in malfunctioning of the normal cellular metabolism. © 2021 by Begell House, Inc.

Advanced modalities in the treatment of prostate cancer, an important health problem among men, draw attention to the field of cancer immunotherapy. Although the immune system holds promise for prostate cancer treatment, the biological processes associated with prostate cancer cells and immune cells have not yet been fully elucidated. The prostate cancer microenvironment is a complex system containing a large number of immune cells. Among multiple immune cells, natural killer (NK) cells have enormous potential in targeting various cancer cells. NK cells, which are an important part of the innate immune system, play a very important role as a first-line defense against prostate cancer cells. Various strategies have been adopted by researchers to increase the efficiency of NK cells. However, there are many difficulties to increase the therapeutic relevance of NK cells. This review will provide a perspective on the phenotypic and functional properties of NK cells, molecular links between NK cells and prostate cancer cells, and the therapeutic and prognostic significances of NK cells in both advanced hormone naïve prostate cancer and castration-resistant prostate cancer. © 2021 Elsevier Inc. All rights reserved.

mTOR is a member of the PI3K/Akt/mTOR signaling pathway that participates in cell growth, proliferation, protein synthesis, transcription, angiogenesis, apoptosis and autophagy. mTOR and MAPK pahways are two important key signal pathways which are related to each other. We investigated the role of mTOR and other signaling molecules in rat ovaries and uteruses in stages of the estrous cycle. Young adult female rats were divided into four groups as proestrous, estrous, metestrous and diestrous according to vaginal smears. Immunohistochemical staining of mTORC1, IGF1, PI3K, pAKT1/2/3, ERK1 and pERK1/2 was performed and pAKT1/2/3 and ERK1 were also analyzed using western blotting on ovarian and uterine tissue samples. According to our results, PI3K/Akt/mTOR and ERK/pERK showed an increase in the rat ovulation period. When all the groups were evaluated the immunoreactivities for all of the antibodies were detected in the oocytes, granulosa and theca cells, corpus luteum and stroma of ovary and lamina propria, surface and glandular epithelium of uterus with the strongest observed with anti-ERK1 antibody and then with a decreasing trend with anti-mTORC1, anti-pAkt1/2/3, anti-IGF1, anti-PI3K and anti-pERK1/2 antibodies in the proestrus and estrus stages. Differently from other parts of the ovary, highest antibody expression in the corpus luteum was observed in the metestrous stage. Moreover, the existence of pAKT1/2/3 and ERK1 proteins was confirmed with the Western blotting technique. We suggest that mTOR and mTOR-related ERK signaling molecules may participate in the rat ovulation process.

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

Unlike low tumorigenic bulk tumor cells (non-CSCs), cancer stem cells (CSCs) are a subset of tumor cells that can self-renew and differentiate into different cancer subtypes. CSCs are considered responsible for tumor recurrence, distant metastasis, angiogenesis, and drug or radiation resistance. CSCs also are resistant to apoptosis. Zoledronic acid (ZA) is a third generation bisphosphonate that reduces cell proliferation and exhibits anti-tumor effects by inducing cell death in some malignancies; however, the effects of ZA on CSCs are unclear. We investigated the anti-cancer effects of ZA on two epithelial cancer cell lines, prostate DU-145 and breast MCF7, focusing primarily on induction and activation of apoptosis. Cluster of differentiation (CD) 133+/CD44+ prostate CSCs and CD 44+/CD24 breast CSCs were isolated from the DU-145 human prostate cancer and MCF-7 human breast cancer cell lines, respectively, using FACSAria flow cytometry cell sorting. CSCs and non-CSCs were exposed to increasing concentrations of ZA for 24, 48 and 72 h to determine the IC50 dose. Annexin-V assay for detecting cell death and cell cycle was performed using the Muse™ Cell Analyzer. Prostate CSCs and non-CSCs were assayed by quantitative reverse transcription PCR (qRT-PCR) array for detecting 84 key apoptosis related genes. Gene regulation at the protein level was investigated by immunofluorescence. ZA caused a dose- and time-dependent decrease in cell viability. Treatment with ZA resulted in a concomitant increase in apoptosis and cell cycle arrest at S-phase in CSCs. Significant over/under-expressions were detected in seven of the genes of ZA-treated DU-145 CSCs cells. Expressions of CASP9, CASP4, BAX and BAD genes increased, while the expressions of BIRC3, BIRC2 and BCL2 genes decreased. In the DU-145 non-CSCs, five genes exhibited changes in gene expression after ZA treatment, two exhibited increased expression (CASP7 and BAD) and three exhibited decreased expression (BIRC3, BIRC2 and BCL2). ZA caused cell death of drug resistant breast MCF-7 and prostate DU-145 cancer stem cells by activating apoptosis. ZA can facilitate the intrinsic pathway of apoptosis in human prostate CSCs by down-regulating anti-apoptotic genes and up-regulating pro-apoptotic genes. ZA may be an effective therapeutic agent for targeting chemoresistance in CSCs.

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.

Despite numerous advances in the field of cancer cell biology, the origin of cancer cells and the systemic disruptions in the cancer process of normal cells have not yet been fully identified. However, we certainly know that cancer cells carry genetic and molecular similarities of the early embryonic development indicating an interesting relationship between “birth” and “death.” During the embryonic development process, the software of system codes to determine cell fate has been designed. Then, cells containing embryonic residues can be directed as a normal cell, cancer cell, or a stem cell. The transcription factor Yin Yang 1 (YY1) has been reported to play critical roles in many biological processes, such as the embryonic development, cell fate specification, regulation of pluripotency, and oncogenesis. In view of these activities, YY1 has been implicated in both the embryonic development and oncogenesis by acting as a bridge linking these two important biological processes. Therefore, in this review, we primarily focus on YY1 investigations linking the embryonic development, oncogenesis, and stem cell biology. © 2021 Elsevier Inc. All rights reserved.

Objective: To investigate the gene expression profile of CSCs and to explore the key pathways and specific molecular signatures involved in the characteristic of CSCs. Materials and methods: CD133+ /CD44+ CSCs and bulk population (non-CSCs) were isolated from DU-145 cells using fluorescence-activated cell sorting (FACS). We used Illumina HumanHT-12 v4 Expression to investigate gene expression profiling of CSCs and non-CSCs. Protein–protein interaction (PPI) network analysis was performed using the STRING database. Biomarkers selected based on gene expression profiling were visually analyzed using immunofluorescence staining method. An image analysis program, ImageJ®, was used for the analysis of fluorescence intensity. Results: In microarray analysis, we found that many ribosomal proteins and translation initiation factors that constitute the mTOR complex were highly expressed. PPI analysis using the 33 genes demonstrated that there was a close interaction between ribosome biogenesis, translation, and mTOR signaling. The fluorescence amount of mTOR and MLST8 were higher in CSCs compared to non-CSCs. Conclusions: The increase in a number of genes associated with ribosome biogenesis, translation, and mTOR signaling may be important to evaluate prognosis and determine treatment approach for prostate cancer (PCa). A better understanding of the molecular pathways associated with CSCs may be promising to develop targeted therapies to prolong survival in PCa. © 2019, Federación de Sociedades Españolas de Oncología (FESEO).

High recurrence and metastatic behavior patterns are the most important reasons for the failure of treatment strategies in patients with colon cancer. Cancer stem cells (CSCs), which are considered root of cancer, are thought to be associated with therapy resistance, relapse, and metastasis, and, therefore, targeting CSCs rather than the bulk population may be an effective approach. In cancer studies, there is an increasing interest in close friendship between epithelial-mesenchymal transition (EMT) and CSCs. Triptolide (TPL) isolated from Chinese herb Tripterygium wilfordii has important effects on the prevention of migration and metastasis as well as cytotoxic effect against cancer cells. The potential lethal efficacy of TPL on CSCs that is highly resistant to the drug is an unsolved mystery. Fundamentally, the present study basically aims to find answers to two questions: (a) is it possible to target colon CSCs with TPL? and (b) what are the mechanisms underlying TPL's potential to eliminate CSCs? Cytotoxic effects of TPL on CSCs were evaluated by WST-1 and Muse count and viability assays. Apoptosis assay and cell-cycle analysis were performed to investigate the inhibitory effect of TPL. Moreover, the effects of TPL on spheroid formation capacity, migration, and EMT processes, which are associated with CSC phenotype, were also investigated. The results revealed that TPL triggered cell death and apoptosis and altered cell cycle distribution. Moreover, TPL significantly reduced the snail slug and twist expressions associated with EMT. TPL has been shown to be effective in colon CSCs by in vitro experiments, and it might be a highly effective agent against colon cancer has been implicated in need of supporting in vivo and clinical studies.

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 α-helices), the abundance of nucleic acids (DNA/RNA), altered nucleic acid conformation and carbohydrate composition. Interestingly, macromolecules containing the C[double bond, length as m-dash]O groups and negatively charged molecules having the COO

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.

Glioblastoma multiforme (GBM) is the most malignant and aggressive primary human brain tumors. The regulatory pathways of apoptosis are altered in GBMs, leading to a survival advantage of the tumor cells. Thus, identification of target molecules, which are effective in triggering of the cell death mechanisms in GBM, is an essential strategy for therapeutic purposes. Glycogen synthase kinase-3 (GSK-3) plays an important role in apoptosis, proliferation and cell cycle. This study focused on the effect of GSK-3 inhibitor IX in the GBM cells. Apoptosis induction was determined by Annexin-V assay, multicaspase activity and immunofluorescence analyses. Concentration-dependent effects of GSK-3 inhibitor IX on the cell cycle were also evaluated. Moreover, the effect of GSK inhibitor on the cellular biomolecules was assessed by using ATR-FTIR spectroscopy. Our assay results indicated that GSK-3 inhibitor IX induces apoptosis, resulting in a significant increase in the expression of caspase-3 and caspase-8 proteins. Cell cycle analyses revealed that GSK-3 inhibitor IX leads to dose-dependent G2/M-phase cell cycle arrest. Based on the FTIR data, treatment of GBM cells causes dysregulation in the carbohydrate metabolism and induces apoptotic cell death which was characterized by the spectral alterations in nucleic acids, an increment in the lipid amount with disordering state and compositional changes in the cellular proteins. These findings suggest that GSK-3 inhibitor IX exhibits anti-cancer effects by inducing apoptosis and changing biomolecular structure of membrane lipids, carbohydrates, nucleic acids and proteins, and thus, may be further evaluated as a potential effective candidate agent for the GBM combination therapies.

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.

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: 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).

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.

Diabetes mellitus (DM) affects many organs including kidney. Tyrosine kinase can cause hypoglycemia and sunitinib is an inhibitor of tyrosine kinase. We investigated the possible effects of sunitinib on the kidney of streptozotocin (STZ) induced type 1 diabetic mice. We used 28 CD 1 type male mice divided into four groups of seven. Type 1 diabetes was induced by injection of STZ. Group 1 was the untreated control. Group 2 comprised non-diabetic mice + sunitinib. Both groups 1 and 2 exhibited normal blood glucose levels. Group 3 comprised STZ treated diabetic mice + saline. Group 4 were diabetic mice + sunitinib treatment. Kidneys were removed after 8 weeks. The immunoreactivities of vimentin, E-cadherin and S100 were assessed. Immunostaining of vimentin, E-cadherin and S100 was located in both the glomeruli and tubules of the kidney. We found that the number of vimentin and E-cadherin positive glomeruli and tubules were increased after sunitinib treatment compared to saline treated diabetic mice. The number of vimentin labeled tubules was decreased in the sunitinib treated group compared to diabetic + saline groups. Differences in the number of S100 positive tubules and glomeruli between groups 3 and 4 were not statistically significant. The effect of sunitinib on experimental diabetic mice appears to be related to levels of vimentin, E-cadherin and S100 in the glomeruli and tubules of the kidney, and sunitinib may protect against renal damage from DM.

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 surface epithelium (OSE) has the characteristics of a stem cell and the potential for differentiation. Previous studies on this subject have succeeded in deriving oocytes from OSE stem cells, leading to the belief that OSE could be used for infertility treatment.Each rat (n = 10) was subjected to zinc and/or progesterone injection for 5 days after conception. After a 6-day implantation period, ovarian tissues were removed and comprehensive immunohistochemical analysis of stem cell markers was conducted: Sox2, Klf4, Oct3/4, c-Myc, CD117, CD90, SSEA-1 and Notch pathway analysis; Notch1, Jagged1, and Delta1 in the OSE and ovarian stromal cells were evaluated after treatment with zinc, progesterone, or both.Progesterone moderately affected Sox2 expression (p0.001), while zinc application strongly affected Klf4 and Oct3/4 and immunoreactivity (p0.001). CD90 immunoreactivity was decreased in the OSE and stroma of the progesterone group (p = 0.006) compared with the zinc (p = 0.244) and zinc/progesterone groups (p = 0.910). On the other hand, SSEA-1 showed moderate staining in the OSE and weak staining in stromal cells in animals treated with zinc (p = 0.727), progesterone (p = 0.626), and zinc/progesterone (p = 0.371), with no differences compared with control. Zinc application affected Notch pathway immunoreactivity, with a significant increase in Notch1 (p = 0.0015) and Jagged1 (p0.001).The expression of putative stem cell markers in the OSE was verified and stem cell receptor activity was raised in the OSE and ovarian stromal cells by zinc and progesterone. Thus, this increased expression allows the therapeutic use of zinc and progesterone in ovary-related infertility and brings a different perspective to reproductive medicine.

The menopause has a negative effect in the skin. Melatonin affects skin functions and structures through actions mediated by cell-surface and putative-nuclear receptors expressed in skin cell. We have therefore determined the effects of melatonin treatment on stem cell in the epidermis and extracellular matrix related molecules in the dermis the skin of postmenopausal rats. A total of 45 female rats were divided into 5 groups: control group, group A [ovariectomy (OVX)], group B (OVX +10 mg/kg/day melatonin), group C (OVX +30 mg/kg/day melatonin), group S (sham operated + 10 mg/kg/day melatonin). Ventral skin samples were excised at 12th week after ovariectomy. Hematoxylin-eosin, periodic acid- methylamine silver, elastic van Gieson staining techniques were used to measure histomorphometrically the thickness of elastic fibers and basement membrane, depths of the epidermis, dermis, and subcutaneous fat layer. Immunohistochemical staining methods were used for fibroblast growth factor β (FGF β), collagen type I, fibronectin, β-catenin, c-kit, c-Myc evaluation. Epidermal thickness, subcutaneous fat layer, and elastic fibers were significantly decreased in group C, and there was a significant increase after melatonin treatment. Although there was no difference in dermal thickness of group C, melatonin also significantly increased the dermal thickness. High FGF β, type I collagen, fibronectin, β-catenin, c-Myc immunoreactivity developed following melatonin in all groups. Thus melatonin treatment of postmenopausal rats was mostly due to the decrease of stem cell and extracellular matrix-related molecules in the skin.

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.