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Amaç: Bu çalışmanın amacı, P. chamaepeuce (L.) Less. (Asteraceae)’nin topraküstü kısımlarından hazırlanan n-hekzan, kloroform ve metanol ekstrelerinin antimikrobiyal ve sitotoksik aktivitelerinin araştırılmasıdır.Gereç ve Yöntem: Ekstrelerin antimikrobiyal aktiviteleri, standart bakteri kökenleri (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Salmonella enterica, Pseudomonas aeruginosa, Bacillus subtilis ve Streptococcus pneumoniae) ve mantar kökenleri (Candida albicans ve C. parapsilosis) üzerinde mikrodilüsyon metodu ile araştırılmış ve minimum inhibitör konsantrasyon (MİK) değerleri belirlenmiştir. Ekstrelerin sitotoksik aktivitesi, WST-1 reaktifi kullanılarak hücre proliferasyon analiz yöntemi ile HeLa, U2OS, PC3, MCF-7 ve A549 olmak üzere farklı kanser hücre hatları üzerinde incelenmiştir.Sonuç ve Tartışma: Bitkiden elde edilen n-hekzan, kloroform ve metanol ekstrelerinin MİK değerlerinin S. aureus, E. faecalis ve B. subtilis’e karşı 250-1000 µg/mL konsantrasyon aralığında olduğu saptanmıştır. n-hekzan ve kloroform ekstrelerinin tüm kanser hücre hatlarında ise değişen düzeyde sitotoksik aktiviteye sahip oldukları belirlenmiştir (IC50: 21.0-67.1 μg/mL).

The heterogeneity of cancer strongly suggests the need to explore additional pathways to target. As cancer cells have increased proteotoxic stress, targeting proteotoxic stress-related pathways such as endoplasmic reticulum stress is attracting attention as new anticancer treatments. One of the downstream responses to endoplasmic reticulum stress is endoplasmic reticulum-associated degradation (ERAD), a major degradation pathway that facilitates proteasome-dependent degradation of unfolded or misfolded proteins. Recently, SVIP, an endogenous ERAD inhibitor, has been implicated in cancer progression, especially in glioma, prostate, and head and neck cancers. Here, the data of several RNA-seq and gene array studies were combined to evaluate the SVIP gene expression analysis on a variety of cancers, with a particular focus on breast cancer. SVIP was found to be overexpressed in primary breast tumors compared to normal tissues correlated well with its promoter methylation status and genetic alterations. Similarly, immunoblotting analysis showed that SVIP was expressed significantly higher in breast cancer cell lines compared to non-tumorigenic epithelial cell line. On the other hand, the expression of the key proteins of gp78-mediated ERAD did not exhibit such a pattern. Interestingly, silencing of SVIP enhanced the proliferation of p53 wt MCF7 cells but not p53 mutant T47D cells, however increased migration ability of both cell lines. Interestingly, SVIP expression is high in primary breast tumors but low in breast metastatic tumors. This correlates well with a lower probability of survival of breast cancer patients with lower SVIP expression compared to the patients with overexpressed SVIP. Overall, our data revealing the differential expression and function of SVIP on breast cancer cell lines together within silicodata analysis suggest that SVIP may have complex functions in breast cancer progression and has the potential to be a therapeutic target for breast cancer.

Background/aim: Nonsteroidal antiinflammatory drugs (NSAIDs) including diclofenac, naproxen, ibuprofen, acetylsalicylic acid, and acetaminophen have been shown to have antimicrobial effects on various microorganisms. The aim of this study was to investigate the antibacterial effects of NSAIDs on Staphylococcus aureus. Materials and methods: Susceptibilities of S. aureus strains to NSAIDs with or without antimicrobials (moxifloxacin, vancomycin, ciprofloxacin, clindamycin, and gentamicin) were determined using the microdilution method and disk diffusion test. Expression levels of genes in the presence of drugs were investigated by real-time quantitative RT-PCR (qRT-PCR), and immunoblotting analysis was performed for staphylococcal protein A (SpA). Results: Our results showed that all NSAIDs were active against S. aureus strains with MIC values ranging from 195 mu g/mL to 6250 mu g/ mL. NSAIDs increased the antibiotic susceptibility of the strains, and diclofenac was found to be more effective than the other drugs. Drugs showed different effects on expression levels of virulence factor and/or regulatory genes. Immunoblotting analysis of SpA protein was mostly in accordance with qRT-PCR results. Conclusion: The regulatory/virulence factor genes and proteins of S. aureus investigated in this study may be reasonable targets for these drugs, and we suggest that the data may contribute to the field of infection control and antimicrobial resistance., Scientific and Technological Research Council of Turkey (TUBITAK) [114S821]; Ege University BAP Coordinatorship [13ECZ007]; Ege University EBILTEM [2015BIL003], This work was supported by the Scientific and Technological Research Council of Turkey (TUBITAK, 114S821), Ege University BAP Coordinatorship (13ECZ007) and Ege University EBILTEM (2015BIL003). The authors thank Prof. Dr. Mine Hosgor-Limoncu, Asst. Prof. Yalcin Erzurumlu, and Pharm. Recep Ilhan, and would like to acknowledge the assistance of Ege University/Faculty of Pharmacy/Pharmaceutical Sciences Research Center (FABAL) and Izmir Institute of Technology/Biotechnology and Bioengineering Application and Research Center (BIYOMER).

In the present study, a sensitive electrochemical aptamer-based biosensing strategy for human non-small cell lung cancer (NSCLC) detection was proposed using nanofiber-modified disposable pencil graphite electrodes (PGEs). the composite nanofiber was comprised of polyacrylonitrile (PAN) and polypyrrole (PPy) polymers, and fabrication of the nanofibers was accomplished using electrospinning process onto PGEs. Development of the nanofibers was confirmed using scanning electron microscopy (SEM). the high-affinity 5 '-aminohexyl-linked aptamer was immobilized onto a PAN/PPy composite nanofiber-modified sensor surface via covalent bonding strategy. After incubation with NSCLC living cells (A549 cell line) at 37.5 degrees C, the recognition between aptamer and target cells was monitored by electrochemical impedance spectroscopy (EIS). the selectivity of the aptasensor was evaluated using nonspecific human cervical cancer cells (HeLa) and a nonspecific aptamer sequence. the proposed electrochemical aptasensor showed high sensitivity toward A549 cells with a detection limit of 1.2 x 10(3)cells/mL. the results indicate that our label-free electrochemical aptasensor has great potential in the design of aptasensors for the diagnostics of other types of cancer cells with broad detection capability in clinical analysis., Ege University, Graduate School of Natural and Applied Science [18/FBE/001], The authors acknowledge financial support from Ege University, Graduate School of Natural and Applied Science, Project Coordinator (18/FBE/001). We also acknowledge the technical support from the Pharmaceutical Sciences Research Center (FABAL), Faculty of Pharmacy, Ege University.

Background: The complexity of cancer biology and the development of chemotherapy resistance are two main obstacles to cancer treatment and necessitate novel anticancer molecules that target different cell death pathways. Modulation of Endoplasmic Reticulum (ER) stress and subsequent activation of the Unfolded Protein Response (UPR) has been proposed as a potential chemotherapeutic target, as prolonged ER stress can lead to cell death via apoptosis or necrosis. Objective: The present study aims to evaluate the molecular mechanism underlying the cytotoxic activity of selected urea and carbohydrazide derivatives. Methods: Cell proliferation assays were performed on HeLa, Capan-1, MCF-7, HCC-1937, and MRC-5 cell lines by WST-1 assay. The expression levels of selected ER stress, autophagy, and apoptosis marker proteins were compared by immunoblotting to characterize the underlying mechanism of cytotoxicity. Flow cytometry was used to detect apoptosis. Results: Of the tested cytotoxic compounds, 3a, 4a, 5a, 6a, and 1b dramatically and 5b moderately increased ER stress-related CHOP protein levels. Interestingly, 5b but not 3a, 4a, 5a, 6a, or 1b increased the expression of pro-apoptotic proteins such as cleaved PARP-1 and cleaved caspase-3 and -7. The flow-cytometry analysis further confirmed that the cytotoxic activity of 5b but not the other compounds is mediated by apoptosis, demonstrated by a significant increase in the percentage of late apoptotic cells (7-AAD/annexin V double-positive cells). Conclusion: Our results suggest that changing a substituent from trifluoromethyl to nitro in urea and carbohydrazide core structure alters the cell death mechanism from apoptosis to an apoptosis-independent cell death pathway. This study shows an example of how such simple modifications of a core chemical structure could cause the induction of divergent cell death pathways., Scientific and Technical Research Council of Turkey (TUBITAK) Research Fund [215S112], This study was supported by the Scientific and Technical Research Council of Turkey (TUBITAK) Research Fund under project number 215S112.

Purpose Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death worldwide with. lack of effective systemic chemotherapy. In this study, we aimed to evaluate the value of ATPase family AAA domain-containing protein 2 (ATAD2) as a biomarker and potential therapeutic target for HCC. Methods The expression of ATAD2 was tested in different HCC patient cohorts by immunohistochemistry and comparative transcriptional analysis. The co-expression of ATAD2 and proliferation markers was compared during liver regeneration and malignancy with different bioinformatics tools. The cellular effects of ATAD2 inactivation in liver malignancy was tested on cell cycle, apoptosis and colony formation ability as well as tumor formation using RNA interference. The genes affected by ATAD2 inactivation in three different HCC cell lines were identified by global gene expression profiling and bioinformatics tools. Results ATAD2 is overexpression is closely correlated with HCC tumor stage. There was gradual increase from dysplasia, well differentiated and poorly differentiated HCC, respectively. We also observed transient upregulation of ATAD2 expression during rat liver regeneration in parallel to changes in Ki-67 expression. ATAD2 knockdown resulted in apoptosis and decreased cell survival in vitro and decreased tumor formation in some HCC cell lines. However, three other HCC cell lines tested where not affected. Similarly, gene expression response to ATAD2 inactivation in different HCC cell lines was highly heterogeneous. Conclusions ATAD2 is a potential proliferation marker for liver regeneration and HCC. It may also serve as a therapeutic target despite heterogeneous response of malignant cells.

Ekiz, Guner/0000-0001-5741-1343; Bedir, Erdal/0000-0003-1262-063X; Ballar, Petek/0000-0002-6189-1818, WOS: 000499741400007, PubMed: 31713424, Biotransformation of Astragalus sapogenins (cycloastragenol (1) and astragenol (2)) by Astragalus species originated endophytic fungi resulted in the production of five new metabolites (3, 7, 10, 12, 14) together with 10 known compounds. the structures of the new compounds were established by NMR spectroscopic and HRMS analysis. Oxygenation, oxidation, epoxidation, dehydrogenation, and ring cleavage reactions were observed on the cycloartane (9,19-cyclolanostane) nucleus. the ability of the compounds to increase telomerase activity in neonatal cells was also evaluated. After prescreening studies to define potent telomerase activators, four compounds were selected for subsequent bioassays. These were performed using very low doses ranging from 0.1 to 30 nM compared to the control cells treated with DMSO. the positive control cycloastragenol and 8 were found to be the most active compounds, with 5.2- (2 nM) and 5.1- (0.5 nM) fold activations versus DMSO, respectively. At the lowest dose of 0.1 nM, compounds 4 and 13 provided 3.5- and 3.8-fold activations, respectively, while cycloastragenol showed a limited activation (1.5-fold)., Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [114Z958], This project was supported by the Scientific and Technological Research Council of Turkey (TUBITAK, Project No. 114Z958). We are very grateful to Bionorm Natural Products for donating cycloastragenol and astragenol, and special thanks are due to the NMR spectrometer operator, A. Anwarulhaque, of Prince Sattam bin Abdulaziz University, Al-Kharj, Saudi Arabia.

A series of novel N-aryl-1-(4-sulfamoylphenyl)-5-(thiophen-2-yl)-1H-pyrazole-3-carboxamides was synthesized and examined as inhibitors of cytosolic (human) hCA I and hCA II, and cancer-related transmembrane hCA IX and hCA XII isoenzymes. AC2 was the most selective inhibitor towards cancer-related hCA IX while AC8 and AC9 selectively inhibited hCA XII over off-target isoenzymes. Anticancer effects of the compounds were evaluated towards human oral squamous cell carcinoma (OSCC) cell lines, human mesenchymal normal oral cells, breast (MCF7), prostate (PC3), non-small cell lung carcinoma cells (A549), and non-tumoral fetal lung fibroblast cells (MRC5). Compounds moderately showed cytotoxicity towards cancer cell lines. Among others, AC6 showed cell specific cytotoxic activity and induced apoptosis in a dose-dependent manner without a significant change in the cell cycle distribution of MCF7. These results suggest that pyrazole-3-carboxamides need further molecular modification to increase their anticancer drug candidate potency., Scientific and Technological Research Council of Turkey (TUBITAK) [219S076], We gratefully acknowledge financial support from The Scientific and Technological Research Council of Turkey (TUBITAK, Project no: 219S076) .

A series of novel 4-aminobenzensulfonamide/carboxamide derivatives bearing naphthoquinone pharmacophore were designed, sythesized and evaluated for their proteasome inhibitory and anti-proliferative activities against human breast cancer cell line (MCF-7). The structures of the synthesized compounds were confirmed by spectral and elemental analyses. The proteasome inhibitory activity studies were carried out using cell-based assay. The antiproteasomal activity results revealed that most of the compounds exhibited inhibitory activity with different percentages against the caspase-like (C-L, beta 1 subunit), trypsin-like (T-L, beta 2 subunit) and chymotrypsin-like (ChT-L, beta 5 subunit) activities of proteasome. Among the tested compounds, compound 14 bearing 5-chloro-2-pyridyl ring on the nitrogen atom of sulfonamide group is the most active compound in the series and displayed higher inhibition with IC50 values of 9.90 +/- 0.61, 44.83 +/- 4.23 and 22.27 +/- 0.15 mu M against ChT-L, C-L and T-L activities of proteasome compared to the lead compound PI-083 (IC50 = 12.47 +/- 0.21, 53.12 +/- 2.56 and 26.37 +/- 0.5 mu M), respectively. The antiproliferative activity was also determined by MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay in vitro. According to the antiproliferative activity results, all of the compounds exhibited cell growth inhibitory activity in a range of IC50 = 1.72 +/- 0.14-20.8 +/- 0.5 mu M and compounds 13 and 28 were found to be the most active compounds with IC50 values of 1.79 +/- 0.21 and 1.72 +/- 0.14 mu M, respectively. Furthermore, molecular modeling studies were carried out for the compounds 13, 14 and 28 to investigate the ligand-enzyme binding interactions. (c) 2020 Elsevier Masson SAS. All rights reserved., Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [116S300]; Ege UniversityEge University [14ECZ042], This study was supported by grants from The Scientific and Technological Research Council of Turkey (TUBITAK, Project Number:116S300) and Ege University (Project Number: 14ECZ042). The authors thank also to the Pharmaceutical Sciences Research Centre (FABAL) at Ege University, Faculty of Pharmacy for spectral and elemental analyses of the compounds.

Small molecules which activate distinct cell death pathways have promising high potential for anticancer drug research. Especially, regulated necrosis draws attention as an alternative cell death mechanism to overcome the drug resistance. Here, we report that a new semisynthetic saponin analogue (AG-08) triggers necrotic cell death with unprecedented pathways. AG-08-mediated necrosis depends on enhanced global proteolysis involving calpains, cathepsins, and caspases. Moreover, AG-08 generates several alterations in lysosomal function and physiology including membrane permeabilization, redistribution toward the perinuclear area, and lastly excessive tubulation. As a consequence of lysosomal impairment, the autophagic process was abolished via AG-08 treatment. Collectively, in addition to its ability to induce necrotic cell death, which makes AG-08 a promising candidate to cope with drug resistance, its unique activity mechanisms including autophagy/lysosome impairment and enhancement of proteolysis leading a strong death capacity emphasizes its potential for anticancer drug research., Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [118S709, 109S345], This project was supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Project No: 118S709) as a continuation of a prior project (TUBITAK, Project No: 109S345).

Proteostasis regulates key cellular processes such as cell proliferation, differentiation, transcription, and apoptosis. The mechanisms by which proteostasis is regulated are crucial and the deterioration of cellular proteostasis has been significantly associated with tumorigenesis since it specifically targets key oncoproteins and tumor suppressors. Prostate cancer (PCa) is the second most common cause of cancer death in men worldwide. Androgens mediate one of the most central signaling pathways in all stages of PCa via the androgen receptor (AR). In addition to their regulation by hormones, PCa cells are also known to be highly secretory and are particularly prone to ER stress as proper ER function is essential. Alterations in various complex signaling pathways and cellular processes including cell cycle control, transcription, DNA repair, apoptosis, cell adhesion, epithelial-mesenchymal transition (EMT), and angiogenesis are critical factors influencing PCa development through key molecular changes mainly by posttranslational modifications in PCa-related proteins, including AR, NKX3.1, PTEN, p53, cyclin D1, and p27. Several ubiquitin ligases like MDM2, Siah2, RNF6, CHIP, and substrate-binding adaptor SPOP; deubiquitinases such as USP7, USP10, USP26, and USP12 are just some of the modifiers involved in the regulation of these key proteins via ubiquitin-proteasome system (UPS). Some ubiquitin-like modifiers, especially SUMOs, have been also closely associated with PCa. On the other hand, the proteotoxicity resulting from misfolded proteins and failure of ER adaptive capacity induce unfolded protein response (UPR) that is an indispensable signaling mechanism for PCa development. Lastly, ER-associated degradation (ERAD) also plays a crucial role in prostate tumorigenesis. In this section, the relationship between prostate cancer and proteostasis will be discussed in terms of UPS, UPR, SUMOylation, ERAD, and autophagy.

Proteostasis regulates key cellular processes such as cell proliferation, differentiation, transcription, and apoptosis. the mechanisms by which proteostasis is regulated are crucial and the deterioration of cellular proteostasis has been significantly associated with tumorigenesis since it specifically targets key oncoproteins and tumor suppressors. Prostate cancer (PCa) is the second most common cause of cancer death in men worldwide. Androgens mediate one of the most central signaling pathways in all stages of PCa via the androgen receptor (AR). in addition to their regulation by hormones, PCa cells are also known to be highly secretory and are particularly prone to ER stress as proper ER function is essential. Alterations in various complex signaling pathways and cellular processes including cell cycle control, transcription, DNA repair, apoptosis, cell adhesion, epithelial-mesenchymal transition (EMT), and angiogenesis are critical factors influencing PCa development through key molecular changes mainly by posttranslational modifications in PCa-related proteins, including AR, NKX3.1, PTEN, p53, cyclin D1, and p27. Several ubiquitin ligases like MDM2, Siah2, RNF6, CHIP, and substrate-binding adaptor SPOP; deubiquitinases such as USP7, USP10, USP26, and USP12 are just some of the modifiers involved in the regulation of these key proteins via ubiquitin-proteasome system (UPS). Some ubiquitin-like modifiers, especially SUMOs, have been also closely associated with PCa. on the other hand, the proteotoxicity resulting from misfolded proteins and failure of ER adaptive capacity induce unfolded protein response (UPR) that is an indispensable signaling mechanism for PCa development. Lastly, ER-associated degradation (ERAD) also plays a crucial role in prostate tumorigenesis. in this section, the relationship between prostate cancer and proteostasis will be discussed in terms of UPS, UPR, SUMOylation, ERAD, and autophagy., Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [SBAG-108S056/114S062]; Ege University internal funds; BAGEP Award of the Science Academy; Pfizer-TurkeyPfizer; COST Action (PROTEOSTASIS)European Cooperation in Science and Technology (COST) [BM1307]; COST (European Cooperation in Science and Technology)European Cooperation in Science and Technology (COST), Work by PBK is supported by the Scientific and Technological Research Council of Turkey (TUBITAK, SBAG-108S056/114S062), Ege University internal funds, BAGEP Award of the Science Academy with funding supplied by Pfizer-Turkey, COST Action (PROTEOSTASIS BM1307), and by COST (European Cooperation in Science and Technology).

Amaç: Bu çalışmada, endoplazmik retikulum-aracılı yıkım yolağındakiretrotranslokasyon basamağının anahtar proteini olan p97/VCP ve etkileşimpartnerleri olan Ufd1 ve Npl4 proteinlerinin NF-κB yolağı üzerine etkileriprostat kanser hücre hattında incelendi.Gereç veYöntem: p97/VCP, Ufd1 ve Npl4 ifadeleri RNA interferans RNAi(RNA interferans) teknolojisi ile LNCAP hücrelerinde susturuldu ve NF-κBaktivitesi ikili lusiferaz yöntemi ile NF-κB yolağı proteinlerinin ifadesi iseimmünoblotlama ile değerlendirildi.Bulgular: p97/VCP, Ufd1 ve Npl4 ifadelerinin susturulması LNCaP hücrelerinde NF-κBaktivitesini anlamlı olarak azaltmış ve IκBα protein seviyesini arttırırkenfosforile NF-κB ve fosforile IκBα seviyelerini azaltmıştır.Sonuç: Retrotranslokasyon kompleks üyelerinin susturulması ile LNCaPhücrelerinde NF-κB aktivitesinin azalmasının NF-κB inhibitörü olan IκBαseviyesinin artmasına bağlı olduğu düşünülmektedir. Bulgularımız p97/VCP veetkileşim partnerleri Ufd1-Npl4 proteinlerinin prostat kanserinde NF-κByolağının düzenlenmesinde önemli bir etken olabileceğini önermektedir., Aim: In this study, the effects of p97/VCP and itsinteracting partners Ufd1 and Npl4 proteins, which function at theretrotranslocation step of endoplasmic reticulum-associated degradation wereinvestigated on the NF-κB pathway in the prostate cancer cell line.Materials and Methods: The expressions of p97/VCP,Ufd1 and Npl4 were silenced in LNCaP cells using RNAi (RNA interference)technology. NF-κB activity was evaluated by dual luciferase assay system andthe expressionof NF-κB pathway proteins were investigated using immunoblotting.Results: Silencing of expression of p97/VCP, Ufd1 and Npl4significantly diminished NF-κB activity and increased the level of IκBα proteinwhile decreased phosphorylated NF-κB and phosphorylated IκBα levels in LNCaPcells. Conclusion: The decrease NF-κB activityupon silencing the expressions of retrotranslocation complex members in LNCaPcells might be associated with the increased level of NF-κB inhibitor IκBα. Ourdata suggests that p97/VCP and its interacting partners might be importantfactors on the regulation of NF-κB pathway in the prostate cancer.

Hypericum genus holds an important place in the flora of Turkey with 46 endemic species out of its naturally growing 96 species. H. perforatum is the most popular and common species. This species has been traditionally used in pain control, wound healing and especially as antidepressant. Its anticancer effects have been lately attracting much attention. To investigate and compare the antiproliferative effects of extracts of the samples collected from eleven different "States" of Turkey. the antiproliferative effect of extracts was determined by WST-1 method on HeLa, U2OS, HCC-1937 cancer cells and MRC-5 noncancer fibroblast cells. Furthermore, cell death mechanism of the extracts was analyzed by investigating the cleavage level of caspase-7 and PARP-1, which are hallmarks of apoptosis and also the levels of LC3-II and p62 protein levels as markers of autophagy. Only HP10 (collected from Cankm-Ilgaz) was found to induce apoptosis both using immunoblotting and flow cytometry analysis, while almost all of the extracts cause the induction of autophagy in HeLa cells at their IC50 values. HP10 coded sample was collected from the transition zone between the Black Sea and Central Anatolian Regions, the collection area with highest altitude. According to our results different climatic features effect plant contents as well as their therapeutic effects. Moreover, the amount of compounds that have anticancer activity in the plant undergo a change with altitude.

In this study, new dibenzensulfonamides, 7–9, having the chemical structure 4,4′-(5′-chloro-3′-methyl-5-aryl-3,4-dihydro-1′H,H-[3,4′-bipyrazole]-1′,2-diyl)dibenzenesulfonamide were synthesized in five steps to develop new anticancer drug candidates. Their chemical structures were confirmed by 1H NMR, 13C NMR and HRMS spectra. Cytotoxicities of the dibenzensulfonamides were investigated towards HCC1937, MCF7, HeLa, A549 as tumor cell lines and towards MRC5 and Vero as non-tumor cells. Carbonic anhydrase (CAs, EC 4.2.1.1) inhibitory effects of the dibenzensulfonamides 7–9 were also evaluated on the cytosolic human (h) hCA I and II and the tumor-associated hCA IX and XII isoenzymes. Results indicate that both 7 and 8 induced cleavage of poly (ADP ribose) polymerase (PARP), activation of caspases -3, -7 and -9 which are the hallmarks of apoptosis. Meanwhile both compounds induced autophagy in HCC1937 cells which is shown by enhanced expression of LC3 and decreased level of p62 protein. The compounds tested were also effectively inhibited tumor-associated hCA IX and hCA XII isoenzymes in the range of 20.7–28.1 nM and 4.5–9.3 nM, respectively.

The hydrazones of 4-hydrazinylpyridinium bearing alkylphenyl groups on pyridinium nitrogen were synthesized and evaluated for their cytotoxic activity against MCF-7, PC3, U2OS, and HEK293 cell lines by Wst1 cell proliferation assay. Cytotoxic activity results indicated that d derivatives having butylene chain; 4 and 5 series having naphthalene and anthracene ring systems showed high cytotoxic activity (IC50 = 3.27-8.54 μm) on cancer cells. 3d (4-(2-(4-hydroxybenzylidene)hydrazinyl)-1-(4-phenylbutyl)pyridinium bromide) was the most cytotoxic compound with IC50 value of 3.27 μm against MCF-7. The most active derivatives (1d, 2d, 3d, 4, and 5 series) were selected to investigate for the effects on autophagy by analyzing the expression of autophagy marker proteins. The conversion of LC3-I to its lipidated form LC3-II is essential for autophagy and related to autophagosomes. According to our results, all tested compounds except for 3d induced lipidated form LC3-II accumulation. Then, the effects of the compounds on p62 protein level were also analyzed by the immunoblotting as the autophagy inhibition results in accumulation of p62. Further molecular mechanistic studies including morphological analysis and live-death assays indicated that all tested compounds (1d, 2d, 3d, 4, and 5 series) are potent antitumoral molecules and all except for 3d have potential to inhibit autophagic flux.

Background: Poly(ADP-ribosyl)ation (PARylation) catalyzed mainly by PARP1 is a highly regulated posttranslational modification associated with several pathways in cellular physiology and genotoxic deoxyribonucleic acid (DNA) damage response. PAR polymers and PARP enzyme function in DNA integrity maintenance and several PARP inhibitors have entered clinical phase studies for cancer therapies. Material and methods: the effect of bosutinib, a dual Src/ Abl kinase inhibitor, on PARylation was fluorometrically measured. the cytotoxic and chemosensitizing effects were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. the levels of DNA repair proteins and PARP enzyme were examined by immunoblotting. Results: in this study, bosutinib is characterized as a novel PARP inhibitor. Bosutinib inhibited oxidative stress-induced cellular PARylation and nuclear foci formation by downregulating PARP1 levels. Bosutinib was found to be more cytotoxic on Capan1 cells with BRCA2 mutation. Furthermore by acting as a chemosensitizer, bosutinib enhanced the cytotoxicity of doxorubicin (DOXO) and etoposide (ETP) by decreasing phosphorylation of DNA repair enzymes checkpoint kinase 1 (Chk1) and ataxiatelangiectasia mutated (ATM). Conclusion: By inhibition of both PARP and DNA damage checkpoint kinases, bosutinib increased the phospho-H2AX levels, an early indicator of DNA double strand breaks., Amaç: Başlıca PARP1 enzimi ile katalize edilen poli(ADP-ribozil) asyon (PARilasyon), oldukça sıkı regüle edilen bir posttranslasyonal modifikasyon olup hücresel fizyoloji ve genotoksik DNA hasar yanıtındaki çeşitli yolaklar ile ilişkilendirilmiştir. PAR polimerleri ve PARP enzimi, DNA bütünlüğünün korunmasında fonksiyon göstermektedir ve çeşitli PARP inhibitörleri kanser terapisi için klinik faz çalışmalarına dahil edilmiştir. Gereç ve Yöntem: Bir Src/Abl kinaz inhibitörü olan bosutinibin PARilasyona etkisi fluorometrik olarak ölçülmüştür. Sitotoksik ve kemohassaslaştırıcı etki, 3-(4,5dimetilmiazoll- 2-yl)-2,5-difeniltetrazolyum bromit (MTT) yöntemi ile değerlendirilmiştir. DNA onarım proteinleri ve PARP enzim seviyesi immunoblotlama ile incelenmiştir. Bulgular: Bu çalışmada, bosutinib yeni bir PARP inhibitörü olarak karakterize edilmiştir. Bosutinib, PARP1 protein seviyesini azaltarak oksidatif stres ile indüklenmiş hücresel PARilasyonu ve nüklear odak oluşumunu inhibe etmektedir. Bosutinib, BRCA2 mutasyonu içeren Capan1 hücrelerinde daha fazla sitotoksik bulunmuştur. Kemohassaslaştırıcı olarak davranan bosutinib, DNA tamir enzimlerinden Chk1 ve ATM fosforilasyonunu inhibe ederek doksorubisin (DOXO) ve etoposidin (ETP) sitotoksisitesini arttırmaktadır. Hem PARP hem de DNA hasar tamir kinazlarının inhibisyonu ile bosutinib, DNA çift zincir kırıklarının erken bir belirteci olan fosfo-H2AX seviyesini arttırmaktadır. Sonuç: Bu çalışmadaki verilerimiz bosutinibin bir PARP inhibitörü olarak davrandığını ve kemohassaslaştırıcı etkisinin hem PARP hem de DNA tamir proteinlerinin inhibisyonu sonucu gözlendiğini önermektedir.

Incorporation of antioxidants into sunscreens is a logical approach, yet co-delivery of them with UV filters is a challenge. Here, we purposed a combination therapy, in which the chemical UV filter, octyl methoxycinnamate, was accumulated on upper skin while the antioxidant, melatonin, can penetrate deeper layers to show its effects. Melatonin-loaded elastic niosomes and octyl methoxycinnamate Pickering emulsion were prepared separately. Lyophilized elastic niosomes were dispersed into the Pickering emulsion to prepare the proposed combination formulation. The characterization studies of the formulations revealed that elastic niosomes can be prepared with tunable nanometer sizes, whereas Pickering emulsions can encapsulate the UV filter in micrometer-sized droplets. Melatonin-loaded elastic niosomes prepared with Tween80/Span80 mixture were 146 nm with a PI of 0.438, and 58.42% entrapment efficiency was achieved. The mean diameter size of the combination formulation was 27.8 μm. Ex vivo permeation studies revealed that 7.40% of octyl methoxycinnamate and 58% of melatonin were permeated through the rat skin while 27.6% octyl methoxycinnamate and 37% of melatonin accumulated in the skin after 24 h. Cell culture studies with real-time cell analyzer showed that the proposed formulation consist of melatonin-loaded elastic niosomes and octyl methoxycinnamate Pickering emulsion had no negative effect on the cell proliferation and viability. According to α,α-diphenyl-β-picrylhydrazyl free radical scavenging method, the proposed formulation showed as high antioxidant activity as melatonin itself. It is concluded that the proposed formulation would be a promising dual therapy for UV-induced skin damage with co-delivery strategy.

The ubiquitine−proteasome pathway (UPP) plays. [...]

PubMed: 31059704 Polyether compounds, a large group of biologically active metabolites produced by Streptomyces species have been reported to show a variety of bioactivity such as antibacterial, antifungal, antiparasitic, antiviral, and tumour cell cytotoxicity. Since some of these compounds target cancer stem cells and multi-drug resistant cancer cells, this family of compounds have become of high interest. In this study, three polyether-type metabolites (1–3), one of which was a new natural product (3), were isolated from the marine derived Streptomyces cacaoi via antimicrobial activity-guided fractionation studies. As several polyether compounds with structural similarity such as monensin have been linked with autophagy and cell death, we first assessed the cytotoxicity of these three compounds. Compounds 2 and 3, but not 1, were found to be cytotoxic in several cell lines with a higher potency towards cancer cells. Furthermore, 2 and 3 caused accumulation of both autophagy flux markers LC3-II and p62 along with cleavage of caspase-3, caspase-9 and poly (ADP-ribose)polymerase 1 (PARP-1). Interestingly, prolonged treatment of the compounds caused a dramatic downregulation of the proteins related to autophagasome formation in a dose dependent manner. Our findings provide insights on the molecular mechanisms of the polyether-type polyketides, and signify their potency as chemotherapeutic agents through inhibiting autophagy and inducing apoptosis. © 2019 Elsevier B.V. 2012/BİL/028 109S361 This research was supported by grants from TUBITAK (109S361) and EBILTEM (2012/BİL/028) (E.B.).

Poly (ADP-ribose) polymerase (PARP) enzyme catalyzes the transfer of ADP-ribose into target proteins. Therefore, PARP is responsible for DNA repair, cell proliferation, and cell death. In this study, potential PARP enzyme inhibitors were designed and synthesized. The synthesized compounds were elucidated by Fourier-transform infrared spectroscopy, 1 H NMR, 13 C NMR, heteronuclear single-quantum correlation, and mass spectrometry, and their purity was checked via thin-layer chromatography, high-performance liquid chromatography, and elemental analysis. A total of 63 newly synthesized compounds were screened in terms of PARP inhibition by cellular PARylation assay in the HeLa cell line. It was found that 19 compounds significantly inhibited the H 2 O 2 -induced cellular PARylation. The chemosensitizer effect of these compounds in cancer cells treated with doxorubicin (doxo) was investigated. It was found that the combination of potent PARP inhibitors with doxo potentiated a cytotoxic effect, similar to that of olaparib. The results of the molecular docking and absorption, distribution, metabolism, excretion, and toxicity (ADMET) analysis revealed that compound 60 might be classified as a potential PARP inhibitor candidate. Taken together, all of the results suggested that carbohydrazide derivatives could be a promising lead for the treatment for cancer disorders. © TÜBİTAK SAG-C-DRP-100616-0260 Türkiye Bilimsel ve Teknolojik Araştirma Kurumu, TÜBITAK 215S112 This study was supported by the Scientific and Technical Research Council of Turkey (TÜBİTAK) Research Fund under project number: 215S112 and the Marmara University Scientific Research Commission under grant number: SAG-C-DRP-100616-0260.

Cycloastragenol [20(R),24(S)-epoxy-3 beta,6 alpha,16 beta,25-tetrahydroxycycloartane] (CA), the principle sapogenol of many cycloartane-type glycosides found in Astragalus genus, is currently the only natural product in the anti-aging market as telomerase activator. Here, we report biotransformation of 20(27)-octanor-cycloastragenol (1), a thermal degradation product of CA, using Astragalus species originated endophytic fungi, viz. Penicillium roseopurpureum, Alternaria eureka, Neosartorya hiratsukae and Camarosporium laburnicola. Fifteen new biotransformation products (2-16) were isolated, and their structures were established by NMR and HRESIMS. Endophytic fungi were found to be capable of performing hydroxylation, oxidation, ring cleavage-methyl migration, dehydrogenation and Baeyer-Villiger type oxidation reactions on the starting compound (1), which would be difficult to achieve by conventional synthetic methods. in addition, the ability of the metabolites to increase telomerase activation in Hekn cells was evaluated, which showed from 1.08 to 12.4-fold activation compared to the control cells treated with DMSO. Among the compounds tested, 10, 11 and 12 were found to be the most potent in terms of telomerase activation with 12.40-, 7.89- and 5.43-fold increase, respectively (at 0.1, 2 and 10 nM concentrations, respectively)., Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [114Z958], This project was supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Project No: 114Z958) . We are very grateful to Bionorm Natural Products for donating the substrate, and special thanks are due to the NMR spectrometer operator, Anzarulhaque Anwarulhaque, of Prince Sattam bin Abdulaziz University, AlKharj, Saudi Arabia.

WOS: 000450780900010, Objectives: Urea and carbohydrazide derivatives are important compounds exhibiting cytotoxic activities. In this study, a series of new urea and carbohydrazide derivatives containing an pyridine ring were synthesized and evaluated for cytotoxic activity. Materials and Methods: The proposed structures of the synthesized compounds were confirmed using elemental analysis, IR, and H-1-NMR spectroscopic techniques. The cytotoxic potencies of synthesized compounds were determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT) on BRCA mutant-carrying HCC1937 and Capan-1 cell lines, as well as on MCF7, HeLa, and MRC5 cells. Results: 3a, 3b, 3c and 3d showed cytotoxic activity against all cancer cell lines. Conclusion: Our data indicate that compounds 3a-d are more selective to cancer cells compared with nontumoral fibroblasts; however, these compounds are not more potent on HR defective cells with BRCA mutants., TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [215S112]; Marmara University Scientific Research CommissionMarmara University [SAG-C-DRP-100616-0260], This study was supported by TUBITAK 215S112 and Marmara University Scientific Research Commission (project number: SAG-C-DRP-100616-0260).

In this study, non-rigid analogs of thalidomide have been designed in order to develop potentially active, more effective and safer lead molecules for disorders caused or contributed by inflammation. Five different series of acylaminoacetamide compounds were synthesized, and the biological inhibitory potency of the title compounds has been determined by evaluating their effects on COX-2 isoenzyme expression and PGE2 production in A549 (human lung adenocarcinoma) cell lines. Among the studied series, N-[2-(isopropylamino)-2-oxoethyl]isonicotinamide is the most active inhibitory compound on COX-2 isoenzyme expression, and N-[2-oxo-2-(pyrolydine-1-yl)etyl]isonicotinamide is the most active inhibitory compound on the biosynthesis of PGE2. Molecular docking studies and molecular dynamics simulations were also applied to investigate non-covalent interactions of the most active compounds inside the active side of the crystal structure of murine cyclooxygenase 2 (mCOX-2) isoenzyme.

Breast cancer is one of the most common malignancies in women and metastasis is the cause of morbidity and mortality in patients. In the development of metastasis, the matrix metalloproteinase (MMP) family has a very important role in tumor development. MMP-2 and MMP-9 work together for extracellular matrix (ECM) cleavage to increase migration. Tomatine is a secondary metabolite that has a natural defense role against plants, fungi, viruses and bacteria that are synthesized from tomato. In addition, tomatine is also known that it breaks down the cell membrane and is a strong inhibitor in human cancer cells. In this study, it was aimed to evaluate the effect of tomatine on cytotoxicity, apoptosis and matrix metalloproteinase inhibition in MCF-7 cell lines. Human breast cancer cell line (MCF-7) was used as a cell line. In MCF-7 cells, the IC50 dose of tomatine was determined to be 7.07μM. According to the control cells, apoptosis increased 3.4 fold in 48thh. Activation of MMP-2, MMP-9 and MMP-9\NGAL has been shown to decrease significantly in cells treated with tomatine by gelatin zymography compared to the control. As a result, matrix metalloproteinase activity and cell proliferation were suppressed by tomatine and this may provide support in treatment methods.

Isatin, 1H-indoline-2,3-dione, an endogenous compound, is also a synthetically versatile molecule that possesses a diversity of biological activities including anticonvulsant, antibacterial, antifungal, antiviral, anticancer, and cytotoxic properties. Based on the promising cytotoxic activity studies on N-substituted isatin derivatives, a series of 18 derivatives of 2-(2,3-dioxo-2,3-dihydro- 1H-indol-1-yl)-N-phenylacetamide were designed, synthesized, and characterized according to their analytical and spectral data. All of the compounds were evaluated for their cytotoxic activity against MCF7, A549, HeLa, and HEK293 cell lines by real time cell analyzer. Etoposide was used as a standard compound. Briefly, ortho substitutions gave better results compared to meta and para substitutions on the N-phenyl ring and compounds bearing ortho substitutions were more effective on MCF7 cell lines than A549 and HeLa cell lines. 2-(2,3-Dioxo-2,3-dihydro-1H-indol-1-yl)-N- (2-isopropylphenyl)acetamide was the most active compound against all the tested cell lines.

A series of new "hybrid compounds", Mannich base derivatives of planar polycyclic/heterocyclic starting materials, was designed and synthesized. The structures of the compounds were confirmed by spectroscopic methods and elemental analysis. Cytotoxicity of compounds was investigated in three cancer cell lines (PC3, HeLa, and MCF7) and one non-tumoral cell line (293 HEK). We tested the DNA-intercalating capability of the molecules by ethidium bromide (EtBr) fluorescence displacement experiment. Compounds' alkylation potency was investigated via in vitro incubation test using 2-mercaptoethanol, a biomimetic nucleophile. The five of the compounds (7s, 9d, 10b, 11b, 12c) are reported for first time in the literature. Our results suggest that compound 9d has a biological activity close to the reference compound doxorubicin, an intercalating agent in clinical use.