Effects of monensin on caspase-10 mediated apoptosis in glioblastoma multiforme

Glioblastoma multiforme (GBM), en kötü huylu primer merkezi sinir sistemi tümörüdür. Şu anda, GBM için iyileştirici tedavi seçenekleri yoktur ve 5 yıllık hayatta kalma oranı %5’den daha azdır. Monensin, ‘’Streptomyces cinnamonensis’’ den elde edilen antibakteriyal ve antiparazitik etkileri bilinen iyonofor bir antibiyotiktir. Literatürde monensinin GBM hücrelerinin apoptoz mekanizması üzerine etki göster- diği bir çalışmaya rastlanmadığından yapılan bu çalışmanın amacı monensinin U373 GBM hücrelerinde apoptoz aracılı hücre proliferasyonu üzerine etkilerini araştırmaktır. Monensinin U373 hücre canlılığı üzerine etkileri XTT ile apoptoz üzerine etkileri ise RT-PCR ve Annexin V ile araştırılmıştır. Monensinin U373 GBM hücrelerinde IC 50 değeri 48’inci saatte 4 μM olarak bulunmuştur. Monensin U373 GBM hücrele- rinde apoptoz oranında 6 katlık bir artışa neden olmuştur. Bununla birlikte monensin kaspaz-10 gen ekspresyonunu arttırarak apoptozu anlamlı olarak aktive etmiştir. Sunulan çalışma monensinin GBM hücrelerinin kaspaz-10 aracılı apoptoz mekanizması üzerine etkilerini gösteren ilk çalışmadır. Bizim sonuçlarımız monensinin GBM kanserinde güçlü apoptotik etkileri olan terapötik bir antikanser ilaç bileşiği olabileceğini önermektedir.Glioblastoma multiforme (GBM) is the most malignant primary central nervous system tumor. Currently, there are no curative treatment options for GBM and the 5-year survival rate is less than 5%. Therefore, further studies are needed to identify effective markers and thera- peutic targets for GBM treatment. Monensin is an ionophore antibiotic obtained from Streptomyces cinnamonensis with known antibacterial and antiparasitic effects. In the literature, no study has been found that shows an effect of monensin on the apoptosis mechanism of GBM cells. The aim of the present study was to investigate the effects of monensin on apoptosis-mediated cell proliferation in U373 GBM cells. The effects of monensin on U373 cell viability were investigated by XTT and its effects on apoptosis were investigated by RT-PCR and Annexin V. The IC50 value of monensin in U373 GBM cells was 4 μM at 48 hours. Monensin caused a 6-fold increase in the rate of apoptosis in U373 GBM cells. Monensin also significantly triggered apoptosis by increasing caspase-10 gene expression. The present study is the first to demonstrate the effects of monensin on the caspase-10-mediated apoptosis mechanism of GBM cells. Our results suggest that monensin may be a therapeutic anticancer drug compound with potent apoptotic effects in GBM cancer

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

Extract of Calvatia gigantea inhibits proliferation of A549 human lung cancer cells

In this study, in order to investigate the anticancer mechanism of Calvatia gigantea extract, edible mushroom species, which belong to Lycoperdaceae family, changes of CCND1, CCND2, CDK4, p21, Akt, Bax, Bcl-2, p53, caspase-3 and caspase-9 were evaluated in A549 lung cancer cells. Cytotoxic effect of C.gigantea extract was evaluated by using XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5 carboxanilide). The C. gigantea extract was treated in a time and dose dependent manner within the range 25 μg/ml–2 mg/ml to determine the IC50 dose. IC50 dose for C. gigantea extract was detected as 500 μg/ml for 72 h. According to expression results, while CCND1, CCND2, CDK4, Akt and Bcl-2 expression clearly decreased, Bax, p53, caspase-3 and caspase-9 expression clearly increased in the dose group cells (A549 cells treated with 500 μg/ml dose of C. gigantea extract for 72 h). However, there was no change in p21 expression. C. gigantea extract induced cell cycle arrest and apoptosis by decreasing the CCND1, CCND2, CDK4, Akt and Bcl-2 expression and by increasing Bax, p53, caspase-3 and caspase-9 expression in A549 cells. Mushrooms are eukaryotic organisms heavily used because of their supposedly anticancer effect. Many mushroom species have been used for medical purposes, as a result of also having many effects such as antibiotic, antiviral and anticancer effects. It is thought that the C. gigantea extract may be a significant agent for treatment of lung cancer as a single agent or in combination with other drugs. © 2016, Springer Science+Business Media Dordrecht

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)

Explant Culture of Ovarian Tissue

INTRODUCTION: In this study, it was aimed to isolate, reproduce and characterize stromal cells migrating from tissues by creating ovarian tissue explant culture. It is also aimed to create a mixed cell culture (ovarian stromal stem cells and ovarian surface epithelium) with the tissues obtained from different parts of the ovary and to examine the interactions of the cells with each other. METHODS: Explant cultures were formed from ovarian tissues of 4 week old (prepubertal) two female Wistar Albino type rats. Then, the expression of CD29, CD54, CD90 (mesenchymal stem cell surface antigen) and CD45 (hematopoietic stem cell surface antigen) was investigated by performing flow cytometry analysis on proliferating ovarian stromal cells in the 2nd passage (P2). RESULTS: The proliferation abilities and morphological characteristics of the cells in the culture medium were examined by serial passaging. In flow cytometry analysis of isolated ovarian stromal cells, it was determined that they expressed CD54, CD90 and CD45 surface antigens, but did not express CD29 surface antigens. DISCUSSION AND CONCLUSION: In the analysis, we determined that the ovarian stromal cells we isolated and produced in the culture medium expressed hematopoietic and some mesenchymal stem cell markers

Determination of the effects of oleuropein on various cellular pathways in the sh-sy5y neuroblastoma cell line

Nöroblastom beyin dokusu içerisinde hızlı bir Ģekilde yayılabilen ve çocukluk çağının en sık rastlanılan tümör çeĢitlerinden birisidir. Kanser hastalığı her geçen gün geliĢen teĢhis ve tedavi yöntemlerine rağmen hala ölüm nedeni olarak dünyada ilk sıralardadır. Günümüzde kanser tedavisinde cerrahi müdahale, kemoterapi, immünoterapi, gen terapisi, radyoterapi ve bunlara destek amaçlı çeĢitli alternatif tedaviler kullanılmaktadır. Son yıllarda çeĢitli kanser türlerinin tedavisinde kullanılabilecek doğal ve biyolojik etkili bileĢiklerin keĢfedilmesi en popüler araĢtırma konuları arasında yer almaktadır. Bu bileĢiklerden biri olan polifenoller grubuna ait, zeytin yaprağının etken maddesi oleuropeinin antioksidan, anti-mikrobiyal, anti-inflamatuar, anti-hipertansif, anti-kanserojen etkileri olduğu bilinmektedir. Bu çalıĢmanın amacı oleuropeinin SH-SY5Y nöroblastom hücre hattında hücre proliferasyonuna, invazyonuna ve koloni oluĢumu, hücre döngüsü ve apoptoz mekanizmaları üzerine etkisi ve terapötik etkinliğinin in vitro olarak belirlenmesidir. Oleuropeinin hücre canlılığı üzerine etkisi XTT yöntemi ile belirlenmiĢ ve IC50 dozu 48. saate 350 μM olarak bulunmuĢtur. Gerçek Zamanlı PZR sonuçlarına göre oleuropein hücre döngüsü ile iliĢkili genlerden SiklinD1, SiklinD2, SiklinD3, CDK4 ve CDK6 down regülasyonuna ve p53 ve p21'in up regülasyonuna sebep olarak hücre döngüsünü durdurduğu ve ayrıca Bcl-2'nin inhibisyonu, Bax, kaspaz-9 ve kaspaz 3'ün aktive edilmesiyle apoptotik yolağı indüklediği tespit edilmiĢtir. Western blot yöntemi ile Bcl-2 ve SiklinD1'in protein düzeyinde değiĢimleri gösterilmiĢtir. TUNEL testi ile doz grubu hücrelerde apoptotik hücre oranının %36.4±3.27'ye çıktığı bulunmuĢtur. Ayrıca Matrigel-Ġnvazyon testi ile oleuropeinin SH-SY5Y hücrelerinde invazyonu azalttığı belirlenmiĢtir. Koloni formasyon testi ile de koloni oluĢumunu %53.6±4.71 oranında baskıladığı tespit edilmiĢtir. ÇalıĢma sonuçları ile oleuropeinin, nöroblastom tedavisinde potansiyel bir anti-kanserojen ajan olabileceği gösterilmiĢ ve daha detaylı çalıĢmalar için ilk veriler ortaya koyulmuĢtur

Antioxidant activity, phytochemical composition of Andricus tomentosus and its antiproliferative effect on Mia-Paca2 cell line

Plant derived products are widely used in cancer treatment. Gall species has been preferred for treatment various diseases in folk medicine, but there are few studies on the anticancer effects of gall species. The present study reports the antioxidant activity and total secondary metabolites of extracts of A. tomentosus galls. The antioxidant potency of galls was carried out using different in-vitro model systems. Their cytotoxic efficacy on Mia-Paca cell line was also explored. Gall extract was found to contain a large amount of phenolic acids. The extract potently scavenged free radicals including DPPH (IC50 9.56 ± 1.08 µg/mL), ABTS (IC50 18.51 ± 0.25 µg/mL). It can be seen as a potential source of antioxidants according to β-carotene/linoleic acid method (%92.58 ± 0.92) and Phosphomolybdenum assays (104.36 ± 4.95 mgAE/g). Gall extract also posses ability of metal chelating (%40.07 ± 2.30) and Fe3+ (184.01 ± 2.83 mgTE/g) and Cu2+ (89.81 ± 0.96 mgTE/g) reducing activity. According to total secondary metabolites tests, gall extract showed high total phenolic, total flavonoid and total tannin amount. HPLC analysis of the extract suggested it to contain caffeic acid (424.068.479 µg/g), ellagic acid (187.696.132 µg/g). XTT assay revealed gall extract to enhance percent survival of Mia-Paca2 cell line exposed A. tomentosus extracts. The best cytotoxic effect was determined in acetone extracts (IC50: 124.7 µM). Expression of some genes (Bax, Bcl-2, FAS, BID, caspase-3, caspase-8, caspase-9, caspase-10, FADD, TRADD) in the apoptosis pathway was determined to invastigate apoptosis inducing activity. These results indicate that A. tomentosus galls possess potent antioxidant activity, when tested both in chemical as well as biological models. © 2020, Springer Nature B.V

Investigation of anticancer mechanism of oleuropein via cell cycle and apoptotic pathways in SH-SY5Y neuroblastoma cells

Neuroblastoma is one of the most common types of pediatric tumors that can spread quickly in neuronal tissues. Oleuropein which is active compound of olive leaves, belongs to polyphenols group and has antioxidant, anti-microbial, anti-inflammatory, anti-hypertensive and anti-carcinogenic effects. The aim of the study is to determine the therapeutic effects of oleuropein on cell proliferation, invasion, colony formation, cell cycle and apoptotic mechanisms in SH-SY5Y neuroblastoma cell line under in vitro conditions. The effect of oleuropein on cell viability was determined by XTT method. 84 cell cycle control and 84 apoptosis related genes were evaluated by RT-PCR. Effects of oleuropein on apoptosis were researched by TUNEL assay. Protein expressions were determined by western blot analysis. Effects of oleuropein on cell invasion, colony formation and migration were detected by matrigel-chamber, colony formation assay and wound-healing assay, respectively. IC50 value of oleuropein in SH-SY5Y cells was detected as 350 μM at 48th hours. It is determined that oleuropein causes cell cycle arrest by down-regulating of CylinD1,CylinD2,CyclinD3,CDK4,CDK6 and up-regulating of p53 and CDKN2A, CDKN2B, CDKN1A gene expressions. Oleuropein also induces apoptosis by inhibiting of Bcl-2 and activating of Bax,caspase-9 and caspase-3 gene expressions. Apoptotic cell ratio was found 36.4 ± 3.27% in oleuropein dose group. Oleuropein decreased invasion in SH-SY5Y cells and suppressed colony numbers in ratio of 53.6 ± 4.71%.Our results demonstrated that oleuropein can be a therapeutic agent in the treatment of neuroblastoma. © 2016 Elsevier B.V

Assessment of the anticancer mechanism of ferulic acid via cell cycle and apoptotic pathways in human prostate cancer cell lines

Studies on genetic changes underlying prostate cancer and the possible signaling pathways are getting increased day by day, and new treatment methods are being searched for. The aim of the present study is to investigate the effects of ferulic acid (FA), a phenolic compound, on cell cycle, apoptosis, invasion, and colony formation in the PC-3 and LNCaP prostate cancer cells. The effect of FA on cell viability was determined via a 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) method. Total RNA was isolated with Tri Reagent. Expression of 84 genes for both cell cycle and apoptosis separately was evaluated by reverse transcriptase PCR (RT-PCR). Protein expressions were evaluated by Western blot analysis. Furthermore, apoptotic effects of FA were observed with terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) assay. Effects of FA on cell invasion and colony formation were determined using Matrigel chamber and colony assay, respectively. The half maximal inhibitory concentration (IC50) dose of FA was found to be 300 μM in PC-3 cells and 500 μM in LNCaP cells. According to RT-PCR results, it was observed that FA inhibited cell proliferation by increasing the gene expressions of ATR, ATM, CDKN1A, CDKN1B, E2F4, RB1, and TP53 and decreasing the gene expressions of CCND1, CCND2, CCND3, CDK2, CDK4, and CDK6 in PC-3 cells. On the other hand, it was seen that FA suppressed cell proliferation by increasing in the gene expressions of CASP1, CASP2, CASP8, CYCS, FAS, FASLG, and TRADD and decreasing in the gene expressions of BCL2 and XIAP in LNCaP cells. In this study, protein expression of CDK4 and BCL2 genes significantly decreased in these cells. It could induce apoptosis in PC-3 and LNCaP cells. Also, it was observed that FA suppressed the invasion in PC-3 and LNCaP cells. Moreover, it suppressed the colony formation. In conclusion, it has been observed that FA may lead to cell cycle arrest in PC-3 cells while it may cause apoptosis in LNCaP cells. © 2015, International Society of Oncology and BioMarkers (ISOBM)