Antidiabetic exendin-4 activates apoptotic pathway and inhibits growth of breast cancer cells

Exendin-4 is a GLP-1 analog used for the treatment of type 2 diabetes mellitus in its synthetic form. As women with diabetes have higher breast cancer incidence and mortality, we examined the effect of the incretin drug exendin-4 on breast cancer cells. The aim of the study is to investigate anticancer mechanism of exendin-4 in MCF-7 breast cancer cells. Cytotoxic effects of exendin-4 were determined by XTT assay. IC50 dose in MCF-7 cells were detected as 5 μM at 48th hour. Gene messenger RNA (mRNA) expressions were evaluated by real-time PCR. According to results, caspase-9, Akt, and MMP2 expression was reduced in dose group cells, compared with the control group cells. p53, caspase-3, caspase-8, caspase-10, BID, DR4, DR5, FADD, TRADD, PARP, PTEN, PUMA, NOXA, APAF, TIMP1, and TIMP2 expression was increased in dose group cells, compared with the control group cells. Effects of exendin-4 on cell invasion, colony formation, and cell migration were detected by Matrigel chamber, colony formation assay, and wound-healing assay, respectively. To conclude, it is thought that exendin-4 demonstrates anticarcinogenesis activity by effecting apoptosis, invasion, migration, and colony formation in MCF-7 cells. Exendin-4 may be a therapeutic agent for treatment of breast cancer as single or in combination with other agents. More detailed researches are required to define the pathways of GLP-1 effect on breast cancer cells because of the molecular biology of breast cancer that involves a complex network of interconnected signaling pathways that have role in cell growth, survival, and cell invasion. © 2015, International Society of Oncology and BioMarkers (ISOBM)

Investigation of the effects of erianin on proliferation and colony formation of HT29 colorectal cancer cells

Kolorektal kanser, dünyada kanser ilişkili ölümlerin en yaygın dördüncü sebebidir. Erianin antioksidan ve anti-tümör etkilere sahip Dendrobium ekstraktından elde edilen yeni bir dibenzil bileşiğidir. Bu çalışmada, erianinin HT29 kolorektal kanser hücreleri üzerine olan terapötik etkileri araştırılmıştır. Erianinin HT29 hücre canlılığı üzerine etkileri XTT test ile koloni oluşumu üzerine etkileri ise koloni formasyonu ile değerlendirilmiştir. Erianinin HT29 hücrelerinde IC50 değeri 48. saatte 59.05 µM olarak belirlenmiştir. HT29 hücre dizisinde erianin uygulanan grupta koloni sayısı 67±33 iken kontrol grubunda 350±89 olarak hesaplanmıştır. Erianin, HT29 kolorektal kanser hücrelerinde koloni oluşumunu ise anlamlı derecede azaltmıştır. Yapılan çalışmaların sonuçları, erianinin kolorektal kanser tedavisinde doğal elde edilen bir bileşik olarak güvenli, kolay ulaşılabilir ve umut veren terapötik bir ilaç olabileceğini destekler niteliktedir. Gelecekte erianinin kolorektal kanser hücreleri üzerindeki etki mekanizmasını aydınlatacak daha kapsamlı ve çok merkezli desteklenecek ileri düzeyde klinik çalışmalara ihtiyaç vardır.Colorectal cancer (CRC) is the 4th most common cause of cancer-related death in the global world. Erianin, a novel dibenzyl compound in Dendrobium extract, has antioxidative and antitumor activities. In this study, the therapeutic effects of erianin on HT29 colorectal cancer cells were investigated. Effects of erianin on cell viability were evaluated by XTT test. Effects of erianin on colony formation were evaluated by colony formation analysis. IC50 values of Erianin on HT29 cells were determined as 59.05 µM at 48th hour. While the number of colonies in the HT29 cell line was 67±33 in the erianin treated group, it was calculated as 350±89 in the control group. Erianin significantly reduced colony formation in HT29 colorectal cancer cells. The results of the presented studies support that erianin as a natural product in the treatment of colorectal cancer can be a safe, easily accessible and promising therapeutic drug. In the future, more comprehensive and multi-center supported clinical studies are needed to elucidate the mechanism of action of erianin on colorectal cancer cells

Zoledronic acid induces apoptosis via stimulating the expressions of ERN1, TLR2, and IRF5 genes in glioma cells

Glioblastoma multiforme (GBM) is the most common and aggressive brain tumor that affects older people. Although the current therapeutic approaches for GBM include surgical resection, radiotherapy, and chemotherapeutic agent temozolomide, the median survival of patients is 14.6 months because of its aggressiveness. Zoledronic acid (ZA) is a nitrogen-containing bisphosphonate that exhibited anticancer activity in different cancers. The purpose of this study was to assess the potential effect of ZA in distinct signal transduction pathways in U87-MG cells. In this study, experiments performed on U87-MG cell line (Human glioblastoma-astrocytoma, epithelial-like cell line) which is an in vitro model of human glioblastoma cells to examine the cytotoxic and apoptotic effects of ZA. IC50 dose of ZA, 25 μM, applied on U87-MG cells during 72 h. ApoDIRECT In Situ DNA Fragmentation Assay was used to investigate apoptosis of U87MG cells. The quantitative reverse transcription polymerase chain reaction (qRT-PCR) (LightCycler480 System) was carried out for 48 gene expression like NF-κB, Toll-like receptors, cytokines, and inteferons. Our results indicated that ZA (IC50 dose) increased apoptosis 1.27-fold in U87MG cells according to control cells. According to qRT-PCR data, expression levels of the endoplasmic reticulum-nuclei-1 (ERN1), Toll-like receptor 2 (TLR2), and human IFN regulatory factor 5 (IRF5) tumor suppressor genes elevated 2.05-, 2.08-, and 2.3-fold by ZA, respectively, in U87MG cells. Our recent results indicated that ZA have a key role in GBM progression and might be considered as a potential agent in glioma treatment. © 2015, International Society of Oncology and BioMarkers (ISOBM)

Anti-proliferative and anti-invasive effects of ferulic acid in TT medullary thyroid cancer cells interacting with URG4/URGCP

Ferulic acid (4-hydroxy-3-methoxycinnamic acid; FA), a common dietary plant phenolic compound, is abundant in fruits and vegetables. The aim of present study is to investigate the effects of FA on cell cycle, apoptosis, invasion, migration, and colony formation in the TT medullary thyroid cancer cell line. The effect of FA on cell viability was determined by using CellTiter-Glo assay. IC50 dose in the TT cells was detected as 150 μM. URG4/URGCP (upregulated gene-4/upregulator of cell proliferation) is a novel gene in full-length mRNA of 3.607 kb located on 7p13. It was determined that FA caused a decrease in the expression of novel gene URG4/URGCP, CCND1, CDK4, CDK6, BCL2, MMP2, and MMP9, a significant increase in the expression of p53, PARP, PUMA, NOXA, BAX, BID, CASP3, CASP9, and TIMP1 genes in TT human thyroid cancer cell line by using real-time PCR. It was found that FA in TT cells suppressed invasion, migration, and colony formation by using matrigel invasion chamber, wound healing, and colony formation assay, respectively. In conclusion, it is thought that FA indicates anticarcinogenesis activity by affecting cell cycle arrest, apoptosis, invasion, migration, and colony formation on TT cells. © 2015, International Society of Oncology and BioMarkers (ISOBM)

Investigation of the effects of Histone Deacetylase inhibitors (HDACI) on apoptotic pathway in Glioblastoma Multiforme (GBM)

Bu çalışma, PAÜ Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından desteklenmiştir (Proje No: 2015SBE001).Glioblastoma multiforme, genellikle yetişkinlerde görülen merkezi sinir sisteminin en yaygın ve agresif primer beyin tümörüdür. Cerrahi müdahele, radyoterapi ve kemoterapi gibi tedavilere rağmen hastalığın sağ kalım 14.6 aydır. Konvansiyonel kemoterapi düşük kan-beyin bariyeri penetrasyonu, tümör içi heterojenite, intrinsik GBM direnci ve spesifik olmayan toksisite nedeniyle sınırlı etkiye sahiptir. TRAIL normal hücrelere herhangi bir etki yapmadan, kanser hücrelerini hedeflemesi yönünden oldukça ilgi çekici bir ajandır. GBM’in TRAIL-aracılı apoptoza direnç gösterdiği bilinmektedir. Bu nedenle HDACi gibi ajanların TRAIL ile birlikte kombine olarak uygulanması GBM tedavisi için güncel bir stratejidir. Bu çalışmanın amacı, Vorinostat, MS-275, Belinostat ve Romidepsin HDACi’lerinden TRAIL-aracılı ölümü en iyi şekilde arttıran 2 HDAC’yi belirlemek ve bu HDACi’lerin GBM hücrelerinde apoptotik yolak üzerindeki etkilerini araştırmaktır. Bu sebeple, LN18, T98G, U87MG ve U373 hücrelerinde TRAIL’in etkisi, HDACi’lerin sitotoksik etkisi ve TRAIL ile kombine uygulaması CellTiter-Glo yöntemi ile belirlenmiş ve GraphPad Prism programı ile IC50 değerleri hesaplanmıştır. Belinostat ve Romidepsin’in TRAIL ile kombine uygulamasının apoptoza olan etkisi akım-sitometri yöntemi ile hesaplanmıştır. Belinostat ve Romidepsin’in pro-apoptotik ve anti-apoptotik genlerin ekspresyonuna olan etkisi kantitatif gerçek-zamanlı PCR yöntemi ile belirlenmiştir. Apoptozda rol oynayan kaspaz-8, -9, Bcl-2 ve Bax proteinlerinin ekspresyonu western blot yöntemi ile belirlenmiştir. Sonuçlarımıza göre; Vorinostat, Romidepsin ve Belinostat GBM hücrelerinde diğer HDACi’lere göre daha fazla sitotoksik etki göstermiştir. Belinostat ve Romidpesin’in diğer HDACi’lere göre TRAIL-aracılı ölümde daha etkili olduğu bulunmuştur. Akım-sitometri analizlerine göre, Belinostat ve Romidepsin’in TRAIL ile kombine uygulamayla apoptozu ciddi şekilde arttırdığı, gerçek-zamanlı PCR sonucuna göre de pro-apoptotik genlerin ekspresyonunu arttırırken, anti-apoptotik genlerin ekspresyonunu azalttığı bulunmuştur. Protein seviyesinde ise anlamlı bir fark görülememiştir. Bu sonuçlar bize, Belinostat ve Romidepsin’in GBM tedavisi için potansiyel birer ajan olduklarını göstermektedir.Glioblastoma multiforme is the most common central nervous system and aggressive brain tumour mainly occur in adults. Despite surgical, radiotherapy and chemotherapy, survival of the disease is 14.6 months. Conventional chemotherapy has limited effect due to blood-brain-barrier, intrinsic heterogeneity of tumor, intrinsic GBM resistance and non-specific toxicity. TRAIL which has no effect for normal cells but targets tumour cells, is a promising agent. It is known that GBM shows resistance to TRAIL-mediated apoptosis. Therefore, combined treatment agents such as HDACi combined with TRAIL is current strategy for GBM therapy. The aim of this study to determine the best two HDACi enhancing the TRAIL-mediated cell death among Vorinostat, MS-275, CBHA, Belinostat and Romidepsin and to invstigate the effects of these HDACis upon apoptotic pathway on GBM. For this reason, effects of TRAIL, cytotoxic effects of HDACis and effects of combined TRAIL treatment on LN18, T98G, U87MG and TRAIL was determined with CellTiter-Glo method and IC50 doses of HDACis were calculated via GraphPad Prism program. Effects of the combination of TRAIL either Belionstat or Romidepsin to apoptosis were calculated by flow-cytometry. Expression changes of pro-apoptotic and anti-apoptotic genes were analyzed by real-time PCR. Additionally, protein expressions of Bcl-2, Bax, caspase-8 and caspase-9 were determined by western blot analysis. According to our results, Vorinotat, Romidepsin and Belinostat have more cytotoxic effect on GBM compared with other HDACis. It is found that compared to other HDACis, Belinostat and Romidepsin are more effective for TRAIL-induced cell death. By flow-cytometry analysis, combined treatment of Belinostat and Romidepsin with TRAIL enhanced apoptosis significantly. According to real-time PCR results, Romidepsin and Belinostat increased pro-apoptotic gene expression but decreased anti-apoptotic expression. There isn’t any siginificant changes on protein levels. This results suggest and indicated that Belinostat and Romidepsin is a potential agent for GBM treatment

Erianin, a promising agent in the treatment of glioblastoma multiforme triggers apoptosis in U373 and A172 glioblastoma cells

Glioblastoma is an aggressive, common and deadly primary intracranial brain tumor in adults. The antitumor activity of erianin, a dibenzyl compound found in Dendrobium chrysotoxum Lindl. extract, has not been previously demonstrated in glioblastoma. We investigated the anticancer activity and underlying mechanisms of erianin in human U373 and A172 glioma cells. The effects of erianin on cell viability, apoptosis, migration and invasion were estimated by the XTT test, the reverse transcription-polymerase chain reaction (RT-PCR), annexin V staining assay protocol for apoptosis, wound healing assay, and Matrigel (R) invasion chamber, respectively. The effective amounts of erianin in U373 and A172 cells were 16 and 64 mu M at 48 h, respectively. Erianin also significantly induced apoptosis by inhibiting B-cell lymphoma 2 (Bcl-2), caspase-8, caspase-9 and tumor necrosis factor receptor type 1-associated DEATH domain protein (TRADD), and activation of caspase-3 and BH3 interacting domain death agonist (BID) gene expression. In addition, erianin significantly increased the number of apoptotic cells in U373 and A172 cells and significantly decreased invasion and migration in U373 and A172 cells. Taken together, our results suggest that erianin may be a new therapeutic anticancer drug component with a potent apoptotic effect and a potential for treating glioblastoma

First note on Holocene coquinite on Thrace (Black Sea) coast of Turkey

In this paper, preliminary results concerning the nature, subsurface structure and age of carbonate-cemented coquinite extending along a 1.5-km long and 40-m wide loose coquina beach on the Thracian (west Black Sea) coast of Turkey are presented. An Electrical Resistivity Tomography (ERT) survey showed that the coquinite has a maximum thickness of about 2 m and comprises seaward dipping buried 20 m-wide slabs under beach materials at the backshore. Cemented by low-magnesian calcite, the coquinite contains various bivalvia, benthic foraminifera, calcareous nannofossils or coccoliths and quartz sands. Diagenesis of the coquinite occurred as the result of sequential cementation, starting with the precipitation of micritic envelops typical of a marine phreatic zone. This early stage was followed by pore-lining cements and bladed rims composed of equal-sized subhedral and anhedral crystals of calcite, suggesting evidence of meteoric phreatic and meteoric vadose environments, respectively. Calibrated values from radiocarbon dating of four bulk samples of bivalvia revealed that the coquinite shells were deposited between 3730 and 2850 years BP. During that period the sea-level was similar to the present at an early stage then dropped to 2 m due to Phanagorian regression (between 3200 and 2200 BP), which is evidenced by consecutive cementation patterns of distinctive origin. (C) 2012 Elsevier B.V. All rights reserved