49 research outputs found
Guidelines for cell viability assays
No full textCapanoglu, Esra/0000-0003-0335-9433; Kamiloglu, Senem/0000-0003-3902-4360; SARI, Gulce/0000-0002-8585-5889Recently, the interest in the application of cell viability assays has been increasing in various fields. Cell viability assays may be broadly classified as (a) dye exclusion assays, (b) colorimetric assays, (c) fluorometric assays, (d) luminometric assays, and (e) flow cytometric assays. Dye exclusion assays include trypan blue, eosin, congo red, and erythrosine B stain assays, whereas 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT), 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS), 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium, monosodium salt (WST-1), 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8), lactate dehydrogenase (LDH), sulforhodamine B (SRB), neutral red uptake (NRU), and crystal violet stain (CVS) assays are among the colorimetric assays. Similarly, resazurin and 5-carboxyfluorescein diacetate acetoxymethyl ester (5-CFDA-AM) assays are based on fluorometric measurements, whereas luminometric assays comprise adenosine triphosphate and real-time viability assays. Major flow cytometric assays include membrane asymmetry, membrane permeability, and mitochondria assays. In this guideline, the mechanisms and the practice of assessment of the most common cell viability assays applied in research labs are discussed in detail. An ideal cell viability assay should be safe, rapid, reliable, efficient, and time- and cost-effective, and should not interfere with the test compound. Overall, it can be concluded that more than one cell viability assay should be applied in order to obtain reliable results
Proteasome Inhibitors in Cancer Therapy and their Relation to Redox Regulation
No full textsahin, ali/0000-0001-5594-1551; Yilmaz, Betul/0000-0003-1762-0284; Daniyal, Muhammad/0000-0002-5663-5211; SARI, Gulce/0000-0002-8585-5889Redox homeostasis is important for the maintenance of cell survival. Under physiological conditions, redox system works in a balance and involves activation of many signaling molecules. Regulation of redox balance via signaling molecules is achieved by different pathways and proteasomal system is a key pathway in this process. Importance of proteasomal system on signaling pathways has been investigated for many years. In this direction, many proteasome targeting molecules have been developed. Some of them are already in the clinic for cancer treatment and some are still under investigation to highlight underlying mechanisms. Although there are many studies done, molecular mechanisms of proteasome inhibitors and related signaling pathways need more detailed explanations. This review aims to discuss redox status and proteasomal system related signaling pathways. In addition, cancer therapies targeting proteasomal system and their effects on redox-related pathways have been summarized
Evaluation of Turkish propolis for its chemical composition, antioxidant capacity, anti-proliferative effect on several human breast cancer cell lines and proliferative effect on fibroblasts and mouse mesenchymal stem cell line
No full textOzdal, Tugba/0000-0001-8344-5336; Capanoglu, Esra/0000-0003-0335-9433; mutlu Altundag, ergul/0000-0002-2377-4047; BOYACIOGLU, DILEK/0000-0002-8160-0619; SARI, Gulce/0000-0002-8585-5889Propolis is an extremely complex resinous natural compound collected by honey bees from various plant sources and exhibits pharmacological and biological properties attributed to the presence of polyphenols. This study examined the total phenolic and flavonoid contents as well as the total antioxidant capacity using 2,2 '-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), cupric-reducing antioxidant capacity (CUPRAC), and ferric-reducing antioxidant power (FRAP) methods. Turkish propolis has very high total phenolic (314.36 +/- 3.65 mg GAE/g propolis) and total flavonoid contents (522.71 +/- 11.45 mg QE/g propolis). The highest antioxidant capacity value was obtained by the CUPRAC method (1184.94 +/- 63.27 mg TE/g propolis). Phenolic profile of Turkish propolis was also determined by high performance liquid chromatography with photodiode array detection (HPLC-PDA) method. The main phenolic compounds identified in Turkish propolis was flavonoids including pinocembrin > chrysin > galangin > pinobanksin > pinostrobin and phenolic acids including caffeic acid > p-coumaric acid > ferrulic acid > t-cinnamic acid. In the present work, anti-proliferative and proliferative effects of propolis extracts were also investigated on two different breast cancer cell lines; MDA-MB-231, UACC-3199 and on two normal cell lines; fibroblasts and mouse mesenchymal stem cell lines. According to the XTT results, Turkish propolis sample showed significant anti-proliferative effect on MDA-MB-231 and UACC breast cancer cell lines. Interestingly, Turkish propolis sample had proliferative effect on both fibroblasts and mouse mesenchymal stem cells. These results suggest that Turkish propolis can be considered as a potent agent on breast cancer treatment for further investigations
Proteomic approach for understanding milder neurotoxicity of Carfilzomib against Bortezomib
No full textYilmaz, Betul/0000-0003-1762-0284; Bergquist, Jonas/0000-0002-4597-041X; Forsberg-Nilsson, Karin/0000-0003-0692-6245; Jannuzzi, Ayse Tarbin/0000-0003-0578-6893; SARI, Gulce/0000-0002-8585-5889; Grune, Tilman/0000-0003-4775-9973; Jung, Tobias/0000-0002-9159-8444; Wicher, Grzegorz/0000-0002-7737-1374The proteasomal system is responsible for the turnover of damaged proteins. Because of its important functions in oncogenesis, inhibiting the proteasomal system is a promising therapeutic approach for cancer treatment. Bortezomib (BTZ) is the first proteasome inhibitor approved by FDA for clinical applications. However neuropathic side effects are dose limiting for BTZ as many other chemotherapeutic agents. Therefore second-generation proteasome inhibitors have been developed including carfilzomib (CFZ). Aim of the present work was investigating the mechanisms of peripheral neuropathy triggered by the proteasome inhibitor BTZ and comparing the pathways affected by BTZ and CFZ, respectively. Neural stem cells, isolated from the cortex of E14 mouse embryos, were treated with BTZ and CFZ and mass spectrometry was used to compare the global protein pool of treated cells. BTZ was shown to cause more severe cytoskeletal damage, which is crucial in neural cell integrity. Excessive protein carbonylation and actin filament destabilization were also detected following BTZ treatment that was lower following CFZ treatment. Our data on cytoskeletal proteins, chaperone system, and protein oxidation may explain the milder neurotoxic effects of CFZ in clinical applications.Short Term Scientific Mission [COST-CM-1001]; Swedish Research Council (SRC) [2015-4870]; Scientific and Technological Research Council of Turkey (TUBITAK) [212T156]; and Okan University International Affairs Office Erasmus Staff Mobility for Training program in SwedenThis study was supported by COST-CM-1001 Short Term Scientific Mission and Okan University International Affairs Office Erasmus Staff Mobility for Training program for the stay of GS in Sweden, by research fundings from Swedish Research Council (SRC) grant 2015-4870 and by The Scientific and Technological Research Council of Turkey (TUBITAK) grant 212T156. We thank Ayse Mine Yilmaz, PhD, Ayca Arslanhan, MSc and Ali Sahin, PhD for their technical support and assistance during experimental procedures. We thank Annika Hohn, PhD for her support during experimental analysis in Department of Molecular Toxicology, German Institute of Human Nutrition Potsdam-Rehbrucke, Germany. We also thank to A. Suha Yalcin, Prof and Tolga Emre, Assoc Prof for English grammar editing
Proteasome Inhibitors in Cancer Therapy and their Relation to Redox Regulation
No full textRedox homeostasis is important for the maintenance of cell survival. Under physiological conditions, redox system works in a balance and involves activation of many signaling molecules. Regulation of redox balance via signaling molecules is achieved by different pathways and proteasomal system is a key pathway in this process. Importance of proteasomal system on signaling pathways has been investigated for many years. In this direction, many proteasome targeting molecules have been developed. Some of them are already in the clinic for cancer treatment and some are still under investigation to highlight underlying mechanisms. Although there are many studies done, molecular mechanisms of proteasome inhibitors and related signaling pathways need more detailed explanations. This review aims to discuss redox status and proteasomal system related signaling pathways. In addition, cancer therapies targeting proteasomal system and their effects on redox-related pathways have been summarized
HBV cccDNA:The Molecular Reservoir of Hepatitis B Persistence and Challenges to Achieve Viral Eradication
No full textHepatitis B virus (HBV) is a major global health issue, with an estimated 254 million people living with chronic HBV infection worldwide as of 2022. Chronic HBV infection is the leading cause of cirrhosis and liver cancer. Current treatment with nucleos(t)ide analogs is effective in the suppression of viral activity but generally requires lifelong treatment. They fail to eradicate the HBV viral reservoir, called covalently closed circular DNA (cccDNA), which replicates in the nucleus of liver cells. The cccDNA serves as the sole template for viral replication, as it generates the pregenomic RNA (pgRNA) necessary for producing new viral genomes. This stable form of viral DNA can reactivate the virus when treatment is stopped. HBV cccDNA is therefore one of the main challenges in curing chronic HBV infections. By targeting steps such as cccDNA formation, capsid assembly, or particle secretion, researchers continue to seek ways to interfere with HBV replication and to reduce its persistence, ultimately to eradicate HBV as a global health problem. This review provides an overview of what is currently known about cccDNA formation and biogenesis and the ongoing efforts to target and eradicate it to cure chronic HBV infections.</p
Proteasomal degradation of TP53INP1 and their relation to methotrexate treatment in prostate cancer cells
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