Two novel series including Schiff bases of the pyrrolizine-5-carboxamides and their Cu(II) complexes were designed, synthesized and analysed using spectral and analytical techniques. The analytical ...results indicated the formation of the complexes in 1:1 or 1:2 (Metal:Ligand) ratio. The geometry around the Cu centers was confirmed to be tetrahedral or octahedral. The cytotoxic activity of the new compounds was evaluated using MCF-7 (human breast adenocarcinoma), A2780 (human ovary adenocarcinoma) and HT29 (human colon adenocarcinoma), in addition to MRC5 (normal human fetal lung fibroblast) cells using the MTT cytotoxicity assay. The Schiff base 12c and the Cu complex 13b were the most active in the two series with IC50 values in the range of 0.14–2.54 μM against the three cell lines. Also, the Cu complex 13e showed excellent activity against HT29 with IC50 = 0.05μM. 7-Cyano-N-(4-methoxyphenyl)-6-((3-phenylallylidene) amino)-2,3-dihydro-1H-pyrrolizine-5-carboxamide (12c) showed high selectivity (6–13 folds) for cancerous cells over normal cells; and it induced marginal increases in the G1 and S phases of MCF-7 cells during cell cycle analysis, while compound 13b increased the MCF-7 Sub-G1 proapoptotic population, and blocked cells in the G2-M phase in a dose dependent manner. The annexin V apoptosis assay revealed the ability of compounds 12c and 13b to increase the early apoptotic MCF-7 cell populations two and three fold, respectively. Furthermore, these findings were supported by data showing that the two compounds (12c and 13b) elicit cytotoxic activity. Taken together, the data presented in this study warrants further in vitro and in vivo investigations.
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•The antitumor activity of novel pyrrolizines and their Cu(II) complexes were evaluated.•The tested compounds showed IC50 values in the range of 0.04–24.66 μM.•The substituted methyl-Cu complex blocked MCF-7 in the G2-M phase of the cell cycle.•The most active compounds increase the early apoptotic MCF-7 cell populations.
The Ccr4-Not complex is one of the major deadenylase factors present in eukaryotic cells. This multi-subunit protein complex is composed of at least seven stably associated subunits in mammalian ...cells including two enzymatic deadenylase subunits: one DEDD (Asp-Glu-Asp-Asp)-type deadenylase (either CNOT7/human Caf1/Caf1a or CNOT8/human Pop2/Caf1b/Calif) and one EEP (endonuclease-exonuclease-phosphatase)-type enzyme (either CNOT6/human Ccr4/Ccr4a or CNOT6L/human Ccr4-like/Ccr4b). Here, the role of the human Ccr4-Not complex in cytoplasmic deadenylation of mRNA is discussed, including the mechanism of its recruitment to mRNA and the role of the BTG/Tob proteins.
This study measured the effects of 5-Fluorouracil (5-FU), calcitriol (VD3), and/or thymoquinone (TQ) single/dual/triple therapies on cell cycle progression, apoptosis, inhibition of the PI3K/AKT/mTOR ...pathway, and oxidative stress against colorectal cancer (CRC).
The HT29, SW480 and SW620 cell lines were treated with 5-FU (50 μM), VD3 (25 μM), and TQ (75 μM), alone or combined for 12 h, prior to cell cycle/apoptosis analyses.
TQ monotherapy had greater anticancer effects to active VD3 or 5-FU, revealing higher expression of p21/p27/PTEN/BAX/Cyto-C/Casp-3 and increased levels of total glutathione, with inhibitions in CCND1/CCND3/BCL-2 and PI3K/AKT/mTOR molecules, alongside higher rates of apoptosis in HT29, SW480 and SW620 cells (P < 0.005 for all markers). Additionally, all combination protocols revealed enhanced modulations of the PI3K/PTEN/Akt/mTOR pathway, higher expression of p21/p27/PTEN/BAX/Cyto-C/Casp-3, and better anti-oxidant effects, than the monotherapies. Although TQ/5-FU and TQ/VD3 co-therapies were better relative to the VD3/5-FU regimen, the best tumoricidal effects were observed with triple therapy in the HT29 and SW480 cell lines, possibly by boosted attenuations of the PI3K/AKT/mTOR oncogenic pathway. In contrast, TQ single treatment was more effective than the triple therapy regimen in metastatic SW620 cells, suggesting that this protocol would be more useful therapeutically in late-stage CRC.
In conclusion, this study is the first to demonstrated enhanced anti-tumorigenic effects for VD3, TQ, and 5-FU triple therapy against CRC cells and could represent the best strategy for treating early stages of malignancy, whereas TQ monotherapy could be a better approach for treating metastatic forms of the disease.
•The anti-cancer effects of TQ were superior to active VD3/5-FU in all cell lines.•TQ + 5-FU co-therapy showed better regulation of the PI3K/Akt/mTOR pathway.•The triple therapy exhibited the best effects in primary cancer cell lines.•However, TQ-monotherapy was better against the metastatic CRC cell line.•Improvements of both approaches could be due to better inhibition of PI3K/Akt/mTOR.
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•Two new series of ethyl benzoate bearing pyrrolizine/indolizine moieties were synthesized.•The new compounds displayed anti-inflammatory and analgesic activities mediated by COX-1/2 ...inhibition.•Potent to moderate cytotoxicity against three (MCF-7, A2780 and HT29) cancer cell lines (IC50 = 0.02–23.35 µM).•Compound 9a induced G1 cell cycle arrest and apoptosis in MCF-7 cells.•The new compounds exhibited nice fitting into COX-1/2 and drug-likeness scores higher than that of ketorolac.
Two new series of ethyl benzoate bearing pyrrolizine and indolizine moieties 8–11 were synthesized and evaluated for their anti-inflammatory and anticancer activities. Among these derivatives, compounds 9a, 10b and 11b displayed in vivo anti-inflammatory and analgesic activity comparable to ibuprofen. The acute ulcerogenicity and histopathological studies revealed better GIT safety profile than ibuprofen. Mechanistic study of these compounds revealed inhibitory activity against COX-1/2 with preferential inhibition of COX-2. Evaluation of cytotoxic activity of the new compounds using MTT assay revealed potent to moderate activity against three human (MCF-7, A2780 and HT29) cancer cell lines (IC50 = 0.02–23.35 µM). Compounds 9a, 10a,b and 11a,b exhibited high cytotoxic selectivity against MCF-7 cells (SI = 4–84). Although the indolizine bearing derivatives 8-11b exhibited higher selectivity to COX-2 than their corresponding pyrrolizine analogs 8-11a, but they were less active and selective against MCF-7 cells. Cell cycle analysis and annexin V-FITC/PI assay revealed G1 cell cycle arrest and induction of apoptosis in MCF-7 cells by compound 9a. The docking study revealed nice fitting of the new compounds into the active site of COX-1/2 with higher affinity to COX-2. Compounds 8–11 displayed drug-likeness score in the range of 0.67–1.56 compared to 1.06 for licofelone. These results suggested that compounds 9a, 10b and 11b could be promising agents in future research as anti-inflammatory and anticancer agents.
This study investigated the chemotherapeutic effects of 5‐fluorouracil (5‐FU), metformin (Met), and/or thymoquinone (TQ) single/dual/triple therapies in the HT29, SW480 and SW620 colon cancer (CRC) ...cell lines. Cell cycle/apoptosis were measured by flow cytometry. The gene and protein expression of apoptosis (PCNA/survivin/BAX/Cytochrome‐C/Caspase‐3) and cell cycle (CCND1/CCND3/p21/p27) molecules, the PI3K/mTOR/HIF1α oncogenic pathway, and glycolysis regulatory enzymes were measured by quantitative‐PCR and Western blot. Markers of oxidative stress were also measured by colorimetric assays. Although all treatments induced anti‐cancer effects related to cell cycle arrest and apoptosis, the triple therapy showed the highest pro‐apoptotic actions that coincided with the lowest expression of CCND1/CCND3/PCNA/survivin and the maximal increases in p21/p27/BAX/Cytochrome‐C/Caspase‐3 in all cell lines. The triple therapy also revealed the best suppression of the PI3K/mTOR/HIF1α pathway by increasing its endogenous inhibitors (PTEN/AMPKα) in all cell lines. Moreover, the lowest expression of lactate dehydrogenase and pyruvate dehydrogenase kinase‐1 with the highest expression of pyruvate dehydrogenase were seen with the triple therapy, which also showed the highest increases in oxidative stress markers (ROS/RNS/MDA/protein carbonyl groups) alongside the lowest antioxidant levels (GSH/CAT) in all cell lines. In conclusion, this is the first study to reveal enhanced anti‐cancer effects for metformin/thymoquinone in CRC that were superior to all monotherapies and the other dual therapies. However, the triple therapy approach showed the best tumoricidal actions related to cell cycle arrest and apoptosis in all cell lines, possibly by enhancing oxidative glycolysis and augmenting oxidative stress through stronger modulation of the PI3K/mTOR/HIF1α oncogenic network.
The HT29, SW480, and SW620 colon cancer cells were treated with 5‐fluorouracil (5‐FU), metformin (Met), and/or thymoquinone (TQ) single/dual/triple therapies for 12 h. The anti‐cancer effects related to cell cycle and apoptosis alongside the expression of the PI3K/mTOR oncogenic pathways, glycolysis regulatory molecules, and markers of oxidative stress were measured. The triple therapy protocol disclosed the best tumoricidal actions in all cell lines.
Accurate gene expression requires the precise control of mRNA levels, which are determined by the relative rates of nuclear (pre-)mRNA synthesis and processing, and cytoplasmic mRNA turnover. A key ...step in mRNA degradation is the removal of the poly(A) tail, which involves several deadenylases including components of the Ccr4-Not complex. Here, we focused on the role of the human paralogues CNOT7 (hCaf1/Caf1a) and CNOT8 (hPop2/Caf1b/Calif), which possess deadenylase activity mediated by DEDD nuclease domains. We show that efficient proliferation requires both subunits, although combined knockdown of CNOT7 and CNOT8 further reduces cell proliferation indicating partial redundancy between these proteins. Interestingly, the function of CNOT7 in cell proliferation partly depends on its catalytic activity. On the other hand, the interaction between CNOT7 and BTG2, a member of the antiproliferative BTG/Tob family involved in transcription and mRNA decay appears less important for proliferation of MCF7 cells, suggesting that CNOT7 does not function solely in conjunction with BTG2. Further analysis of gene expression profiles of CNOT7 and/or CNOT8 knockdown cells underscores the partial redundancy between these subunits and suggests that regulation of several genes, including repression of the antiproliferative genes MSMB and PMP22, by the Ccr4-Not complex contributes to cell proliferation.
This study explored the roles of activins and follistatin in colorectal cancers. Paired malignant and normal colonic tissues were collected from archived paraffin-embedded (
= 90 patients) alongside ...fresh (
= 40 patients) specimen cohorts. Activin β-subunits, follistatin and Smad4 mRNAs and proteins were measured by real-time PCR and immunohistochemistry (IHC). Mature activin-A, -B, -AB and follistatin proteins were measured by ELISA. Cancer tissues having ≤ the 20th percentile of the Smad4 IHC score were considered as low (L-S4) group. The Smad4-intact SW480 and Smad4-null HT29 colon cancer cell lines were treated with activins and follistatin, and cell cycle was analysed by flow cytometry. The cell cycle inducing (CCND1/CCND3) and inhibitory (p21/p27) proteins alongside the survival (survivin/BCL2) and pro-apoptosis (Casp-8/Casp-3) markers were measured by immunofluorescence. Thirty-nine patients had right-sided cancers (30%) and showed higher rates of L-S4 tumours (
= 17; 13.1%) alongside worse clinicopathological characteristics relative to left-sided cancers. The βA-subunit and activin-A increased, whilst βB-subunit and activin-AB decreased, in malignant sites and the late-stage cancers revealed the greatest abnormalities. Interestingly, follistatin declined markedly in early-stage malignant tissues, whilst increased significantly in the advanced stages. All activin molecules were comparable between the early stage right- and left-sided tumours, whereas the late-stage right-sided cancers and L-S4 tumours showed more profound deregulations.
, activin-A increased the numbers of the SW480 cells in sub-G1 and G0/G1-phases, whereas reduced the HT29 cell numbers in the sub-G1 phase with simultaneous increases in the G0/G1 and S phases. The p21/p27/Casp-8/Casp-3 proteins escalated, whilst CCND1/CCND3/BCL2/survivin declined in the SW480 cells following activin-A, whereas activin-A only promoted p21 and p27 alongside reduced CCND3 in the HT29 cells. By contrast, activin-AB increased the numbers of SW480 and HT29 cells in Sub-G1 and G0/G1-phases and promoted the anti-cancer and reduced the oncogenic proteins in both cell lines. In conclusion, activins and follistatin displayed stage-dependent dysregulations and were markedly altered during the advanced stages of right-sided and L-S4 cancers. Moreover, the activin-A actions in CRC could be Smad4-dependent, whereas activin-AB may act as a Smad4-independent tumour suppressor protein.
Cadmium (Cd) is a major environmental pollutant and chronic toxicity could induce nephropathy by increasing renal oxidative stress and inflammation. Although vitamin D (VD) and calcium (Ca) ...prophylactic treatments attenuated Cd-induced cell injury, none of the prior studies measure their renoprotective effects against pre-established Cd-nephropathy.
To measure the alleviating effects of VD and/or Ca single and dual therapies against pre-established nephrotoxicity induced by chronic Cd toxicity prior to treatment initiation.
Forty male adult rats were allocated into: negative controls (NC), positive controls (PC), Ca, VD and VC groups. The study lasted for eight weeks and all animals, except the NC, received CdCl2 in drinking water (44 mg/L) throughout the study. Ca (100 mg/kg) and/or VD (350 IU/kg) were given (five times/week) during the last four weeks to the designated groups. Subsequently, the expression of transforming growth factor-β (TGF-β1), inducible nitric oxide synthase (iNOS), neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), VD synthesising (Cyp27b1) and catabolizing (Cyp24a1) enzymes with VD receptor (VDR) and binding protein (VDBP) was measured in renal tissues. Similarly, renal expression of Ca voltage-dependent channels (CaV1.1/CaV3.1), store-operated channels (RyR1/ITPR1), and binding proteins (CAM/CAMKIIA/S100A1/S100B) were measured. Serum markers of renal function alongside several markers of oxidative stress (MDA/H2O2/GSH/GPx/CAT) and inflammation (IL-6/TNF-α/IL-10) together with renal cell apoptosis and expression of caspase-3 were also measured.
The PC group exhibited hypovitaminosis D, hypocalcaemia, hypercalciuria, proteinuria, reduced creatinine clearance, and increased renal apoptosis/necrosis with higher caspase-3 expression. Markers of renal tissue damage (TGF-β1/iNOS/NGAL/KIM-1), oxidative stress (MDA/H2O2), and inflammation (TNF-α/IL-1β/IL-6) increased, whilst the antioxidants (GSH/GPx/CAT) and IL-10 decreased, in the PC group. The PC renal tissues also showed abnormal expression of Cyp27b1, Cyp24a1, VDR, and VDBP, alongside Ca-membranous (CaV1.1/CaV3.1) and store-operated channels (RyR1/ITPR1) and cytosolic Ca-binding proteins (CAM/CAMKIIA/S100A1/S100B). Although VD was superior to Ca monotherapy, their combination revealed the best mitigation effects by attenuating serum and renal tissue Cd concentrations, inflammation and oxidative stress, alongside modulating the expression of VD/Ca-molecules.
This study is the first to show improved alleviations against Cd-nephropathy by co-supplementing VD and Ca, possibly by better regulation of Ca-dependent anti-oxidative and anti-inflammatory actions.
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•Cd induced abnormal expression of renal VD/Ca2+-regulatory molecules.•The dysregulation of VD/Ca2+-pathways could contribute to nephropathy.•VD and Ca2+ co-supplementation showed enhanced protection than monotherapies.•Co-therapy had better anti-oxidative, anti-inflammatory and calcaemic actions.
In the wake of the COVID-19 pandemic, research indicates that the COVID-19 disease susceptibility varies among individuals depending on their ABO blood groups. Researchers globally commenced ...investigating potential methods to stratify cases according to prognosis depending on several clinical parameters. Since there is evidence of a link between ABO blood groups and disease susceptibility, it could be argued that there is a link between blood groups and disease manifestation and progression. The current study investigates whether clinical manifestation, laboratory, and imaging findings vary among ABO blood groups of hospitalized confirmed COVID-19 patients.This retrospective cohort study was conducted between March 1, 2020 and March 31, 2021 in King Faisal Specialist Hospital and Research Centre Riyadh and Jeddah, Saudi Arabia. Demographic information, clinical information, laboratory findings, and imaging investigations were extracted from the data warehouse for all confirmed COVID-19 patients.A total of 285 admitted patients were included in the study. Of these, 81 (28.4%) were blood group A, 43 (15.1%) were blood group B, 11 (3.9%) were blood group AB, and 150 (52.6%) were blood group O. This was almost consistent with the distribution of blood groups among the Saudi Arabia community. The majority of the study participants (79.6% n = 227) were asymptomatic. The upper respiratory tract infection (P = .014) and shortness of breath showed statistically significant differences between the ABO blood group (P = .009). Moreover, the incidence of the symptoms was highly observed in blood group O followed by A then B except for pharyngeal exudate observed in blood group A. The one-way ANOVA test indicated that among the studied hematological parameters, glucose (P = .004), absolute lymphocyte count (P = .001), and IgA (P = .036) showed statistically significant differences between the means of the ABO blood group. The differences in both X-ray and computed tomography scan findings were statistically nonsignificant among the ABO age group. Only 86 (30.3%) patients were admitted to an intensive care unit, and the majority of them were blood groups O 28.7% (n = 43) and A 37.0% (n = 30). However, the differences in complications' outcomes were statistically nonsignificant among the ABO age group.ABO blood groups among hospitalized COVID-19 patients are not associated with clinical, hematological, radiological, and complications abnormality.
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