Axin1 translocates into the nucleus in hepatocyte growth factor induced epithelial-mesenchymal transition

WOS:000709516300007WOS:000709516300007Axin1 is a scaffold protein belonging to a destruction complex that degrades β-catenin. As an inhibitor of Wnt signaling, Axin1 is one of the most widely studied therapeutic targets in cancer treatment, particularly as targets for Tankyrase inhibitors. Axin1 has long been considered to be uninvolved in transforming growth factor beta induced epithelial-mesenchymal transition (EMT); the latter is known to transform cancer cells to a more aggressive phenotype that show increased cell mobility, drug-resistance and enhanced stem cell properties by remaining in the cytoplasm. In this study, the effects of overexpression of Axin1 on hepatocyte growth factor (HGF) induced EMT in HepG2 cells was investigated through analyses of in-cell western, morphological changes, cell growth curve, population doubling time and immunofluorescence staining. It was observed that Axin1 overexpression in HepG2 cells inhibited cellular growth. HGF treatment of Axin1 overexpressing cells resulted in the induction of EMT with the cells acquiring a fibroblast-like spindle morphology. This is the first report to describe that Axin1 translocated to the nucleus during HGF induced EMT, which is how this protein may exert EMT promoting functions. The result of this observation may rise the questions on the safety of Tankyrase inhibitors which stabilize Axin1 for cancer therapy

P38-beta/SAPK-inhibiting and apoptosis-inducing activities of (E)-4-chloro-2-((3-ethoxy-2-hydroxybenzylidene) amino)phenol

WOS:000534108600001 PubMed ID: 32394730The present study has three purposes; first evaluating cytotoxicity of (E)-4-chloro-2-((3-ethoxy-2-hydroxybenzylidene)amino)phenol (ACES), second deciphering ACES-mediated cellular death mechanism, and third estimating ACES-mediated alterations in the expressions of mitogen-activated protein kinase (MAPK) pathway-related genes. Neutral red uptake assay, cell cycle analysis, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) measurements, caspase 3/7 and 9 activations, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were implemented. IC50 values of ACES-treated five cells were around 4-6 mu g/mL. However, Caco-2 and Huh-7 cells were found to be twofold resistant and fivefold sensitive with IC50 values of 11 mu g/mL and 0.93 mu g/mL, respectively. In this study, it was initially reported that ACES exhibits selective cytotoxicity to Huh-7 cells. In addition, ACES induced apoptosis by nuclear fragmentation, MMP disruption, and intracellular ROS elevation in MCF-7 cells. qRT-PCR experiment indicated the expressions of 30 genes including ATF2, CREB1, MYC, NFATC4 (NFAT3), CCNA1, CCNB1, CCND2, CDK2, CDKN1A (p21CIP1), CDKN1C (p57KIP2), CDKN2A (p16INK4a), CDKN2B (p15INK4b), DLK1, NRAS, CDC42, PAK1, MAP4K1 (HPK1), MAP3K3 (MEKK3), MAP2K3 (MEK3), MAP2K6 (MEK6), MOS, MAPK1 (ERK2), MAPK8 (JNK1), MAPK10 (JNK3), MAPK11 (p38-beta), LAMTOR3 (MP1), MAPK8IP2 (JIP-1), PRDX6 (AOP2), COL1A1, and HSPA5 (Grp78) were downregulated at least 1.5-fold. Moreover, ACES effectively inhibited expressions of genes that code for elements of p38-beta/stress-activated protein kinase (SAPK) pathway. ACES has the potential to be used for the reversal of trastuzumab resistance in breast cancer patients by inhibiting p38/SAPK pathway in MCF-7 cells. Therefore, with the selective cytotoxic, apoptosis-inducing, and p38-beta/SAPK-inhibiting activities, ACES can be utilized for developing a novel anticancer drug

İncir, limon, zeytin ve turp ekzozomlarının MCF-7 hücrelerinde sitotoksik etkisinin araştırılması

WOS:000505765900018With their nano-sized structure, exosomes are involved in a wide variety of cellular processes such as genetic information flow, immune system modulations, intercellular communication, and pathophysiological changes. The use of exosomes are exponentially growing particularly in the areas of identification of biomarkers, development of nanocarriers for effective drug delivery, and vaccine production. In recent years, edible plant derived exosomes gained much interest with their strong antimicrobial activities, modulatory activities on the intestinal stem cells, and the anticancer activities. In this study, the cytotoxic effects of fig, lemon, olive and turnip derived exosomes were investigated through the neutral red uptake assay. It was observed that 100 mu g/mL protein containing lemon and turnip derived exosomes inhibited the cell proliferation significantly, on the other hand, fig and olive derived exosomes did not alter the proliferation of MCF-7 cells. Since the results for the cytotoxic activity of turnip exosomes are original in this research, it was found worthy to emphasis the utility of turnip exosomes for the development of new anticancer agents or new drug delivery nanocarriers.Nano-boyutlu yapılarıyla ekzozomlar genetik bilgi akışı, immun sistem düzenlenmesi ve patofizyolojik değişiklikler gibi çok çeşitli hücresel süreçlere dahil olurlar. Ekzozomların, özellikle biyoişaret tanımlanması, etkin ilaç dağıtımı için nanotaşıyıcıların geliştirilmesi ve aşı üretilmesi alanlarında kullanımları hızla artmaktadır. Son yıllarda yenilebilir bitkilerden türevlenen ekzozomlar yüksek antimikrobiyal etkileri, bağırsak kök hücreleri üzerinde düzenleyici etkileri ve antikanser etkileriyle ilgi çekmektedir. Bu çalışmada incir, limon, zetin ve turp türevli ekzozomların sitotoksik etkileri nötral kırmızısı alım testiyle araştırılmıştır. Protein miktarı 100 µg/mL olan limon ve turp ekzozomlarının hücre çoğalmasını engellediği gözlenirken incir ve zeytin türevli ekzozomların MCF-7 çoğalmasını etkilemediği görülmüştür. Turp ekzozomlarının sitotoksistik aktivitesi sonuçları, bu araştırmada orijinal olduğu için, yeni antikanser ajanlarının veya yeni ilaç dağıtım nanotaşıyıcılarının geliştirilmesi için turp ekzozomları kullanımına vurgu yapmanın değerli olabileceği düşünülmüştü

The transfection of hepg2 cells with truncated β-catenin coding expression vector

β-katenin, Wnt sinyalleşmesinde bir efektör proteinidir. β-katenin mutasyonları; otizm, kolon kanseri, gelişmesel, engel, nörodejenerasyon, baş, tür ve yüz anomalileri gibi çok geniş kapsamlı incelemesinde hedeflenmiştir. Bu, özellikle ekson 3 delesyonu aracılı β-katenin kısalmalarıyla oldukça uzun sürmüştür. Bu büyük boyutlu-katenin proteininin tiplerini ve ekson 3 deyonlu formlarını çok sayıda amaçlarını tamamlayabilecektir. Kontrollü deneyler için, numuneler tip ve ekolayanβ-katenin formları kodlayan ve organizmadan alınan formlarını ifade edebilir. HepG2 alımıβ-katenin proteinleri açısından heterozigot olduğu uzun olduğu için bilindiği için, bu çalışmada, Tip ve ekson 3 delesyonlu ekspresyonlu vektörlerini HepG2 için toplam RNA'sından oluşturmanın kullanılmış olması önceden olmuştur. Bunun için, HepG2 planlanmış RNA izole edilmiş, cDNA parçaları polimeraz zinciru (PZR) ile ilgilidir, ifade vektörleri oluşturularak 5'-uç dizilidir. BLAST incelemesi sonrası hem ekson 3 delesyonlu hem de ucun kodlayanın β-kından pcDNA 3.1 CTNNB1 ifadelerinin E. coli olarak adlandırıldığına göre klonlanmıştır. Görsel bir şekilde, HepG2 ifadesiyle ifadeü ile transfekte edilmiş, β-katenin proteini yüksek düzeydedir. hücreleşme morfolojisi ve populasyon zaman aşımına uğramıştır. Bunun için, HepG2 planlanmış RNA izole edilmiş, cDNA parçaları polimeraz zinciru (PZR) ile ilgilidir, ifade vektörleri oluşturularak 5'-uç dizilidir. BLAST incelemesi sonrası hem ekson 3 delesyonlu hem de ucun kodlayanın β-kından pcDNA 3.1 CTNNB1 ifadelerinin E. coli olarak adlandırıldığına göre klonlanmıştır. Görsel bir şekilde, HepG2 ifadesiyle ifadeü ile transfekte edilmiş, β-katenin proteini yüksek düzeydedir. hücreleşme morfolojisi ve populasyon zaman aşımına uğramıştır. Bunun için, HepG2 planlanmış RNA izole edilmiş, cDNA parçaları polimeraz zinciru (PZR) ile ilgilidir, ifade vektörleri oluşturularak 5'-uç dizilidir. BLAST incelemesi sonrası hem ekson 3 delesyonlu hem de ucun kodlayanın β-kından pcDNA 3.1 CTNNB1 ifadelerinin E. coli olarak adlandırıldığına göre klonlanmıştır. Görsel bir şekilde, HepG2 ifadesiyle ifadeü ile transfekte edilmiş, β-katenin proteini yüksek düzeydedir. hücreleşme morfolojisi ve populasyon zaman aşımına uğramıştır. BLAST incelemesi sonrası hem ekson 3 delesyonlu hem de ucun kodlayanın β-kından pcDNA 3.1 CTNNB1 ifadelerinin E. coli olarak adlandırıldığına göre klonlanmıştır. Görsel bir şekilde, HepG2 ifadesiyle ifadeü ile transfekte edilmiş, β-katenin proteini yüksek düzeydedir. hücreleşme morfolojisi ve populasyon zaman aşımına uğramıştır. BLAST incelemesi sonrası hem ekson 3 delesyonlu hem de ucun kodlayanın β-kından pcDNA 3.1 CTNNB1 ifadelerinin E. coli olarak adlandırıldığına göre klonlanmıştır. Görsel bir şekilde, HepG2 ifadesiyle ifadeü ile transfekte edilmiş, β-katenin proteini yüksek düzeydedir. hücreleşme morfolojisi ve populasyon zaman aşımına uğramıştır.β-catenin is an effector protein in Wnt signaling. β-catenin mutations are reported in the development of many diseases such as autism, colorectal carcinoma, developmental delay, intellectual disability, neurodegeneration, skin, hair and facial anomalies. Exon 3 deletion mediated truncations of the β-catenin associated with these diseases. Therefore understanding the functions of wild type and exon 3 deleted forms of β-catenin may provide an enhancement in the treatment of many diseases. However, to conduct controlled experiments, there could be a demand for the expression vectors that code for wild type and exon 3 deleted forms of β-catenin and originated from the same organism. Since it has long been known that HepG2 cells are heterozygous for β-catenin, in this study, it was found worthy of constructing the expression vectors from the total RNA of HepG2 cells. Then the utility of truncated β-catenin coding pcDNA3.1/CTNNB1 expression vector for upregulation of truncated βcatenin in HepG2 cells was examined. To this end, RNA was isolated from HepG2 cells, cDNA fragments were amplified by polymerase chain reaction (PCR), expression vectors were constructed then sequenced from 5’- prime regions. Following the BLAST analysis, it was concluded that both truncated and wild type β-catenin coding pcDNA3.1/CTNNB1 expression vectors were successfully cloned in E. coli cells. Interestingly, when the parental HepG2 cells were transfected with exon 3 deleted expression vector, β-catenin protein levels were not affected. Moreover, cellular morphology and population doubling time were not significantly altered

Anticancer activities of African medicinal spices and vegetables

WOS:000412468400011

ZnO microparticle-loaded chitosan/poly(vinyl alcohol)/acacia gum nanosphere-based nanocomposite thin film wound dressings for accelerated wound healing

WOS:000484279500001The present study deals with the development of novel ZnO microparticle-loaded chitosan/poly(vinyl alcohol)/acacia gum nanosphere-based nanocomposite thin films through electrospraying and evaluation of their potential use in wound healing applications for skin. ZnO microparticles were synthesized and used as bioactive agents. Morphology, size distribution, structure, and dispersion of the synthesized ZnO microparticles were analyzed by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy (TEM). ZnO microparticles were incorporated into the ternary nanocomposite films by electrospraying technique. Thermogravimetric analyses reveal that incorporation of ZnO microparticles into the nanocomposite structure improves the thermal stability. Mechanical analyses show that tensile strength reaches to the maximum value of 12.75 MPa with 0.6 wt % ZnO content. SEM and TEM micrographs demonstrate that the nanocomposite films consist of nanospheres with nanocapsular structures whose sizes are mostly between 250 and 550 nm. Viability tests established prevailing cellular performance of the fibroblasts on 0.6 wt % ZnO microparticle-loaded nanocomposite films with a viability percentage of 160% compared to the control group. (c) 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 48445.Anadolu University Scientific Research Projects CommitteeAnadolu University [1705F237]This study was funded by the Anadolu University Scientific Research Projects Committee; Project No: 1705F237. The authors thank Mr. Metin Cam, electrical engineer in Department of Electrical and Electronics Engineering, Anadolu University, for his special efforts in constructing the electrospraying system

Cytotoxicity of the methanol extracts of Elephantopus mollis, Kalanchoe crenata and 4 other Cameroonian medicinal plants towards human carcinoma cells

Abstract Background Cancer still constitutes one of the major health concerns globally, causing serious threats on patients, their families, and the healthcare system. Methods In this study, the cytotoxicity of the methanol extract of Elephantopus mollis whole plant (EMW), Enantia chlorantha bark (ECB), Kalanchoe crenata leaves (KCL), Lophira alata bark (LAB), Millettia macrophylla leaves (MML) and Phragmanthera capitata leaves (PCL) towards five human solid cancer cell lines and normal CRL2120 fibroblasts, was evaluated. Extracts were subjected to qualitative chemical screening of their secondary metabolite contents using standard methods. The cytotoxicity of samples was evaluated using neutral red uptake (NR) assay meanwhile caspase activation was detected by caspase-Glo assay. Flow cytometry was used to analyze the cell cycle distribution and the mitochondrial membrane potential (MMP) whilst spectrophotometry was used to measure the levels of reactive oxygen species (ROS). Results Phytochemical analysis revealed the presence of polyphenols, triterpenes and sterols in all extracts. The IC50 values of the best samples ranged from 3.29 μg/mL (towards DLD-1 colorectal adenocarcinoma cells) to 24.38 μg/mL (against small lung cancer A549 cells) for EMW, from 2.33 μg/mL (mesothelioma SPC212 cells) to 28.96 μg/mL (HepG2 hepatocarcinoma) for KCL, and from 0.04 μg/mL (towards SPC212 cells) to 0.55 μg/mL (towards A549 cells) for doxorubicin. EMW induced apoptosis in MCF-7 cells mediated by MMP loss and increased ROS production whilst KCL induced apoptosis via ROS production. Conclusion This study provides evidences of the cytotoxicity of the tested plant extract and highlights the good activity of Elephantopus mollis and Kalanchoe crenata. They deserve more exploration to develop novel cytotoxic drugs

Theoretical and experimental electronic transition behaviour study of 2-((4-(dimethylamino) benzylidene)amino) -4-methylphenol and its cytotoxicity

WOS:000609154800002In this study, a Schiff base (2-((4-(dimethylamino)benzylidene)amino)-4-methylphenol; 7S2) has been synthesized and characterized with 1H, 13C-NMR, IR and elemental analysis methods. An electronic transition behaviour of the Schiff base has been investigated in the different eight solvents by UV-Vis. spectroscopy. The stable geometry of 7S2 has been determined by DFT method with Gaussian09 program (B3LYP/6-311G(2d, p)). 7S2 has been analysed for its target region selection using the SwissTarget program. A docking study has been performed against DHODH protein while its pharmacokinetic properties has been evaluated using SwissADME, Osiris and Molinspiration. 7S2 has important pharmacokinetic and drug-likeness properties against other FDA approved DHODH inhibitors. Further, 7S2 has been tested with the neutral red uptake assay and found cytotoxic for several cancer cell lines, having IC50 values ranged between 18-23 µg/mL. According to the pharmacokinetic properties and cytotoxic activities, we suggest that 7S2 may have a potential as an anticancer drug. © 2020 Elsevier B.V.Anadolu Üniversitesi, Anadolu: 1102F027, 1509F633, 1304F064The authors are grateful to the Anadolu University Scientific Research Projects (Project No. 1509F633 and 1102F027 ) for financial support. The authors also gratefully thank to Anadolu University Scientific Research commission for supporting Gaussian 09 and Gauss View 5.0 programs with the projects (Project No: 1304F064 ). We would like to thank to Anadolu University for providing the opportunity to use the CS ChemBioDraw Ultra 16.0.1.4 for Microsoft Windows program

Two new pterocarpans and a new pyrone derivative with cytotoxic activities from Ptycholobium contortum (N.E.Br.) Brummitt (Leguminosae): Revised NMR assignment of mundulea lactone

Background: Ptycholobium is a genus related to Tephrosia which comprises only three species. Compared to Tephrosia, which has been phytochemically and pharmacologically studied, Ptycholobium species have only few or no reports on their chemical constituents. Moreover, no studies on the cytotoxic activities of its secondary metabolites have been previously documented. Results: From the non polar fractions of the roots bark of Ptycholobium contortum (syn Tephrosia contorta), two new pterocarpans: seputhecarpan C 1 and seputhecarpan D 2 and a new pyrone derivative, ptycholopyrone A 3 were isolated. Alongside, five known compounds identified as 3-?,?-dimethylallyl-4-methoxy-6-styryl-?-pyrone or mundulea lactone 4, glyasperin F 5, seputhecarpan A 6, seputheisoflavone 7 and 5-O-methyl-myo-inositol or sequoyitol 8 were also obtained. Their structures were established by the mean means of spectroscopic data in conjunction to those reported in literature. The NMR assignment of the major compound mundulea lactone 4 is revised in this paper. In addition, the cytotoxicity of the isolated metabolites was evaluated on two lung cancer cell lines A549 and SPC212. 8 was not active while compounds 1, 2, 4-7 displayed antiproliferative effects against the two carcinoma cell lines with IC50 values below 75 ?M. IC50 values below 10 ?M were obtained for 4, 6 and 7 on SPC212 cells. Conclusion: Based on the obtained results, Ptycholobium contortum turns to be a rich source of phenolic metabolites among them some bearing prenyl moieties. This study reports for the first time the isolation of pyrone derivatives 3 and 4 from Ptycholobium genus. The cytotoxicity observed for the isolate is also reported for the first time and shows that 4, 6 and 7 could be chemically explored in order to develop a hit candidate against lung cancer. © 2016 The Author(s)

In vitro cytotoxicity of compounds isolated from Desbordesia glaucescens against human carcinoma cell lines

WOS:000405251800006Malignancies constitute a global health concern and chemotherapy remains the main mode of treatment. The present study was designed to evaluate the cytotoxicity of 8 compounds from Desbordesia glaucescens namely lanosta-7,24-dien-3-one (1), friedelanone (2), friedelanol (3), 3,3'-di-O-methylellagic acid (4), 3,3',4'-tri-0-methylellagic acid (5), ellagic acid (6), 3',4'-di-0-methylellagic acid 4-0-beta-o-glucopyranoside (7) and 3,3'-di-0-methylellagic acid 4'-0-beta-c-xylopyranoside (8) against 4 human carcinoma cell lines and normal CRL2120 fibroblasts. The neutral red uptake (NRU) assay was used for cytotoxicity testing. Caspase-Glo assay, cell cycle analysis, measurements of mitochondrial membrane potential (MMP) and levels of reactive oxygen species (ROS) were used to evaluate apoptosis induction. Compounds 4 and 6 as well as doxorubicin had IC50 values below 45 pM in the four tested cancer cell lines meanwhile other compounds displayed selective activity. The IC50 values ranged from 11.23 mu M (towards breast adenocarcinoma MCF-7 cells) to 44.65 mu M (colon carcinoma Caco-2 cells) for 4, from 14.07 mu M (towards MCF-7 cells) to 77.73 mu M (Caco-2 cells) for 6 and from 0.07 mu M (towards SPC212 cells) to 1.01 mu M (A549 cells) for doxorubicin. Compound 4 induced apoptosis in MCF-7 cells mediated by MMP loss. The constituents of Desbordesia glaucescens and especially ellagic acid (6) and its derivative 4 are potential cytotoxic compounds that deserve more investigations towards developing novel antiproliferative drugs against human carcinoma. (C) 2017 SAAB. Published by Elsevier B.V. All rights reserved.Scientific and Technological Research Counsel of Turkey (TUBITAK); Scientific Research Projects Commission of Anadolu University, Eskisehir, Turkey [1507F563, 1306F110]; International Science Programme, Uppsala University, Sweden (ISP) [KEN-02]V.K. and H.S. are thankful to Scientific and Technological Research Counsel of Turkey (TUBITAK) for 6 months travel grant (to V.K.) and to Scientific Research Projects Commission of Anadolu University, Eskisehir, Turkey for the funding grant 1507F563 (to V.K. and H.S.). A grant for part of this work was also provided by International Science Programme, Uppsala University, Sweden (ISP)-KEN-02 project. IC would like to thank the Scientific Research Projects Commission of Anadolu University, Eskisehir, Turkey for the funding grant (1306F110). Authors are also thankful to Sennur Gorgulti for FACS measurements