Search Results (1,130)

Background: Pancreatic cancer (PC) and colorectal cancer (CRC) are among the most lethal malignancies, with growing evidence pointing to the gut microbiota’s role in their progression. This study aimed to explore the anticancer potential of two microbiota-derived postbiotics, N-acetylcysteine (NAC) and tetrahydro β-carboline carboxylic acid (THC), in targeting some hallmark traits of PC and CRC, both as standalone agents and in combination with standard chemotherapeutics (gemcitabine for PC and 5-fluorouracil (5-FU) for CRC). Methods: Cell viability assays and IC₅₀ determination was assessed using either the Muse™ Count & Viability Kit or the Sulforhodamine B assay; cell death was determined by Annexin V/Propidium Iodide and cell cycle assessed by Propidium Iodide was analyzed by flow cytometry. Results: Here, we found that NAC selectively reduced the viability of PC cells BxPC-3 without triggering apoptosis, while effectively inducing apoptosis in PC cells Panc-1 and in CRC cell lines. THC exhibited stronger anticancer activity, inhibiting proliferation and promoting apoptosis in all tested PC and CRC cells, even at lower concentrations. Combination treatments yielded promising enhancement effects. NAC enhanced the cytotoxicity of gemcitabine in Panc-1 cells through increased apoptosis. NAC, when combined with 5-FU, also increased apoptosis of CRC cells. THC further potentiated gemcitabine’s impact on Panc-1 cells by increasing apoptosis and by inducing cell cycle changes in BxPC-3. In the CRC model, THC co-treatment with 5-FU reduced cell viability and increased apoptosis in all cells. Conclusions: These findings provide preliminary in vitro evidence supporting the potential of integrating microbiota-derived postbiotics with conventional chemotherapy both in PC and CRC. Full article

In this study, three distinct hydrolysates, which are designated Dur-I, Dur-II, and Dur-III, were generated from extrusion-pretreated Durvillaea antarctica biomass by applying viscozyme, cellulase, and α-amylase, respectively. Chemical analyses demonstrated distinct compositional differences among the extracts, whereas FTIR spectra verified the presence of fucose-containing sulfated polysaccharides. Furthermore, NMR analyses revealed pronounced structural variations among the extracts. To investigate neuroprotective properties of Dur-I, Dur-II, and Dur-III, rotenone (Rot) was added to SH-SY5Y cells that had been pretreated with Dur-I/II/III. Here, flow cytometry was employed to assess changes in mitochondrial membrane potential (MMP), Bcl-2 expression, cytochrome c release, caspase-9, -8, and -3 activation, as well as DNA fragmentation. The protective effect of Dur-I/II/III pretreatment of SH-SY5Y cells on the Rot-induced death process was further investigated using cell cycle and annexin V-fluorescein isothiocyanate (FITC)/PI (propidium iodide) double staining analyses. The results reveal that the Rot-induced apoptotic factors were all recovered by the pretreatment of Dur-I/II/III. Moreover, cell cycle and annexin V-FITC/PI double staining analyses also indicated that Dur-I/II/III were capable of protecting SH-SY5Y cells from Rot-induced cytotoxicity. Therefore, these Dur extracts are considered as good candidates for the prevention and treatment of neurodegeneration induced by oxidative stress. Full article

Extracellular vesicles (EVs) are emerging as key factors in maintaining cellular homeostasis, critical mediators of intercellular communication, potential biomarkers, and therapeutic tools. While small EVs have been extensively characterized, the molecular signatures of large EVs (including those generated during regulated cell death pathways) remain poorly defined. Here, we investigated the characteristics of large EVs released during apoptosis and pyroptosis by human monocytic cell lines (THP-1 and U937). Apoptosis was induced by staurosporine and blocked using the pan-caspase inhibitor Q-VD-OPh, whereas pyroptosis was triggered by LPS/nigericin and inhibited with a selective NLRP3 inhibitor. We found that both forms of regulated cell death markedly enhanced the release of large EVs. Both apoptotic and pyroptotic large EVs showed increased Annexin V binding and decreased CD9 expression compared with those released by healthy cells. Large EVs derived from apoptotic and pyroptotic cells exhibited distinct proteomic profiles. Pyroptotic large EVs carried interacting protein networks of RNA-binding proteins and chromatin-associated proteins many of which are known damage-associated molecular patterns or alarmins. In contrast, we found that a subpopulation of apoptotic large EVs was characterized by the presence of dsDNA, and active caspase-3/7. Together, our data shed light on the specific protein cargo of large EVs released by cells during apoptosis and pyroptosis. This study identifies candidate markers of large EVs released by dying cells and may enhance our understanding of the role of EVs in regulated cell death. Full article

Chronic inflammation constitutes a well-established driver of colorectal carcinogenesis, yet the molecular circuitry linking inflammatory receptor signalling to tumour cell survival remains incompletely delineated. Here we demonstrate that the HMG-CoA reductase inhibitors atorvastatin and rosuvastatin modulate inflammatory survival pathways in colorectal cancer cells in a manner consistent with targeted interference with the protease-activated receptor 2 (PAR-2)–extracellular signal-regulated kinase (ERK)–tumour necrosis factor-α (TNF-α) signalling axis. Using lipopolysaccharide-stimulated HT-29 and Caco-2 cells as complementary models of inflammatory colorectal malignancy, we show that both statins selectively attenuate PAR-2 expression at the protein and transcript levels while leaving structurally related PAR-1 unaffected. This pattern of receptor modulation is accompanied by suppression of total ERK1/2 expression, ERK1/2 phosphorylation, and the transcriptional target DUSP6, together with attenuation of TNF-α secretion. Importantly, these signaling shifts are associated with dual apoptotic programs; the extrinsic pathway, reflected by transcriptional upregulation and proteolytic activation of caspase-8; and the intrinsic mitochondrial pathway, evidenced by reciprocal modulation of Bcl-2 family proteins favoring Bax over Bcl-2. Both pathways converge upon activation of executioner caspase-3 and an increase in Annexin V-defined apoptotic fractions, indicating re-engagement of programmed cell death under inflammatory stress. Notably, rosuvastatin consistently demonstrates superior potency across signaling endpoints, achieving comparable biological effects at lower concentrations than atorvastatin. Collectively, these data indicate that clinically deployed statins target the PAR-2–ERK axis and are associated with re-activation of apoptotic pathways in inflammatory colorectal cancer models, while leaving open the possibility that additional statin-responsive networks contribute to their pro-apoptotic effects. This mechanistic framework provides biological plausibility for epidemiologic observations linking statin use with reduced colorectal cancer risk and improved outcomes, and supports further translational evaluation of PAR-2-directed statin strategies in colorectal malignancy. Full article

Glioblastoma (GBM) is a highly aggressive brain tumor characterized by invasive growth, intrinsic drug resistance, and the presence of the blood–brain barrier. All of these features make treatment extremely challenging and underscore the need for developing effective combination strategies and advanced drug delivery systems. This study aimed to develop a bovine serum albumin (BSA) nanoparticle (NP)-based delivery system to overcome the poor bioavailability and pharmacokinetic limitations of two potent anti-tumor agents, all-trans retinoic acid (ATRA) and curcumin (CURC), and to evaluate their antitumor activity in U87-MG GBM cells. Drug-free and ATRA/CURC-loaded BSA-NPs were synthesized using an optimized desolvation method and characterized in terms of particle size, polydispersity index, morphology, drug encapsulation efficiency, and release behavior. The cytotoxic, anti-migratory, and pro-apoptotic effects of the NPs on U87-MG GBM cells were assessed using real-time proliferation and migration assays and Annexin V/PI staining followed by flow cytometry. Collectively, the findings indicated that the co-delivery of ATRA and CURC using BSA-NPs showed enhanced antiproliferative, antimigratory, and pro-apoptotic effects. With its controlled release profile, high loading capacity, and favorable nanoscale dimensions, the ATRA-CURC-BSA–NP system represents a promising nanoplatform for GBM therapy that warrants further in vivo investigation. To the best of our knowledge, this is the first study demonstrating the inhibition of glioblastoma cell growth through the co-delivery of all-trans retinoic acid and curcumin using a bovine serum albumin-based nanoparticle system. Full article

Cotinus coggygria Scop., a member of the Anacardiaceae family, is known for its antiseptic, anti-inflammatory, and antitumor properties. In the present study, the ethyl acetate/water leaf extract of C. coggygria was evaluated for antioxidant and anticancer activities. The extract exhibited strong radical-scavenging potential, effectively neutralizing DPPH, ABTS^•+, and superoxide radicals in a concentration-dependent manner. The cytotoxic effects of the extract on human hepatocellular carcinoma HepG2 cells were also investigated. Flow cytometry revealed significant S-phase cell cycle arrest, while fluorescent microscopy and annexin V-FITC/PI staining demonstrated induction of apoptosis. DNA damage was confirmed by alkaline comet assay. Molecular docking was used to evaluate the binding affinity and inhibitory potential of penta-O-galloyl-β-D-glucose, a representative of gallotannins found in C. coggygria extracts, towards cyclin-dependent kinase 2 and checkpoint kinase 1. A high inhibition ability was demonstrated, which could explain the observed cell cycle block. Collectively, these findings suggest that C. coggygria extract exerts strong antioxidant capacity and selective antiproliferative activity in HepG2 cells. The anticancer effects of C. coggygria extract were associated with DNA damage, cell cycle arrest, disruption of mitochondrial membrane potential, and apoptosis induction. The results show the potential of the herb as a natural therapeutic agent for hepatocellular carcinoma. Full article

Background/Objectives: Cigarette smoke (CS) drives pathogenesis across the spectrum of chronic respiratory disorders, exerting its detrimental effects primarily through oxidative stress and programmed cell death. Scutellarein (Scu), a botanical-origin flavonoid enriched in respiratory therapeutics-oriented Chinese medicinal herbs, demonstrates established anti-inflammatory applications. This study systematically evaluated the protective roles of Scu against CS-induced lung injury and explored the underlying mechanisms. Methods: Subacute CS-exposed mice were used to evaluate the therapeutic effects of Scu on lung injury. Immunofluorescence and quantitative PCR were used to examine the expression levels of junctional proteins and proinflammatory mediators. Apoptotic cell death was quantified using Annexin V-FITC/7-AAD staining. Transepithelial electrical resistance and dextran permeability assay were used to access the barrier integrity in alveolar epithelial MLE-12 cells. Western blotting was used to detect the changes in the signal pathway. Results: In CS-exposed mice, Scu administration dose-dependently reduced histopathological scores, pulmonary edema, changes in the alveolar structure, and inflammatory cell infiltration. In MLE-12 cells, Scu significantly suppressed cigarette smoke condensate (CSC)-induced inflammatory mediators, oxidative stress, caspase-3 activation, and apoptosis and preserved CSC-suppressed tight junction protein expression and barrier disruption. Scu also rescued CSC-altered expression levels of Hrk, Ecscr, and Myo5b and mitigated the CSC-suppressed PI3K/AKT/mTOR pathway. Conclusions: Scu alleviates CS-induced subacute lung injury through its antioxidant, anti-apoptotic effects to maintain epithelial barrier integrity likely via the mitigation of the CSC-suppressed PI3K/AKT/mTOR pathway. Full article

Cervical cancer remains a prominent cause of cancer-related mortality among women worldwide because of chronic infection with high-risk human papillomavirus (HPV) and disparate access to prevention and treatment. The current research evaluates the anticancer activity of Gypenoside XVII, a bioactive saponin of Gynostemma pentaphyllum, in HeLa cells as a model of cervical cancer. MTT, Annexin V-PI, and Hoechst 33342 assays showed dose-dependent growth inhibition with typical apoptotic morphology. Flow cytometry revealed G₀/G₁ cell-cycle arrest, while pathway interrogation revealed participation of mitochondrial and death-receptor cascades, in agreement with caspase-9 and caspase-8 activation, respectively. Collectively, these findings position Gypenoside XVII as a natural-product bioactive with potential both as an anticancer lead and as a functional-food ingredient, deserving of further preclinical development. Full article

Introduction: Incomplete polymerization of in vivo composite resins (CR) poses a significant problem, with monomer-to-polymer conversion rates ranging from around 60 to 75%. Furthermore, oxygen exposure hampers polymerization in the surface layers. This research aims to evaluate the autophagy-inducing potential of three types of CRS and to explore the role of the Akt/mTOR–autophagy–apoptosis crosstalk in composite resin-induced autophagy. The study uses human gingival fibroblasts and three composite materials (M1 and M2, which are 3D printed, and M3, which is milled). Materials and Methods: SEM analysis was performed on the dental materials, and cells kept in contact for 24 h were subjected to tests including the following: MTT, LDH, NO, immunological detection of proteins involved in autophagy and apoptosis, as well as immunofluorescence tests (Annexin V and nucleus; mitochondria and caspase 3/7; detection of autophagosomes). Results: The results showed statistically significant decreases in cell viability with M1 and M2, linked to increases in cytotoxicity and oxidative stress (LDH and NO). Using multiplex techniques, significant increases in glycogen synthase kinase 3 beta (GSK3b) protein were observed in both M1 and M2; a decrease in mTOR (mechanistic target of rapamycin) expression was noted in M1 and M3. Immunofluorescence tests revealed an increase in Annexin V across all materials studied, and an increase in autophagosomes in M1 and M2, whereas a decrease was observed in M3. Conclusions: The relationship between apoptosis and autophagy is highly complex, indicating they may occur sequentially, coexist, or be mutually exclusive. Understanding this complex interplay can help in designing new 3D-printing protocols and monomer compositions to prevent autophagy imbalance. Full article

To investigate the regulatory effects of miR-16-5p and miR-142-3p on inflammation and autophagy in human corneal epithelial cells (HCEpiCs) exposed to hyperosmotic stress, a key pathogenic condition in dry eye disease, HCEpiCs were cultured under NaCl-induced hyperosmotic conditions (450 mOsm, 24 h) and transfected with miR-16-5p or miR-142-3p mimics. Expression of inflammatory cytokines (IL-1β, IL-6, TNF-α, IRAK1), autophagy-related genes (ATG5, Beclin-1, ATG16L1, p62), and apoptotic markers (Bax, Bcl-2, caspase-3) was analyzed by qRT-PCR and Western blot. Reactive oxygen species (ROS), autophagic vesicles, and apoptosis were evaluated using DCFH-DA, DAPRed, and Annexin V assays. The expression levels of antioxidant proteins (SOD1, catalase, NRF2) were also measured. Hyperosmotic stress induces marked inflammatory activation and excessive autophagy in HCEpiCs, accompanied by increased ROS generation and apoptosis. Overexpression of miR-16-5p or miR-142-3p significantly attenuated these effects by suppressing NF-κB-mediated cytokine expression and downregulating ATG5 and ATG16L1 expression, while restoring p62 expression. Both miRNAs reduced oxidative stress and COX-2 expression, enhanced antioxidant defenses, and normalized the expression of apoptotic markers. miR-16-5p and miR-142-3p are important regulators of inflammation and autophagy under hyperosmotic stress. Our findings suggest that modulating intracellular miR-16-5p and miR-142-3p levels in corneal epithelial cells may represent a potential approach to protect the ocular surface under hyperosmotic stress, although their systemic roles in autoimmune dry eye require further clarification. Full article

Background/Objectives: Cervical cancer remains a major global health challenge, and treatment resistance limits the long-term success of chemotherapy. Drug repurposing strategies offer new opportunities for improving therapeutic outcomes by combining existing agents with established chemotherapeutics. Sitagliptin, a DPP-4 inhibitor commonly used in type 2 diabetes, has recently gained attention for its potential anticancer effects. This study aimed to investigate the cytotoxic, apoptotic, and anti-metastatic effects of sitagliptin and doxorubicin, individually and in combination, on human cervical cancer cells (HeLa), and to determine whether their combined use exerts a synergistic anticancer effect. Methods: HeLa cells were treated for 48 h with increasing concentrations of sitagliptin, doxorubicin, or their combination. Cell viability was assessed using the MTT assay. Apoptosis was evaluated by Annexin V-FITC/PI staining and caspase-8/9 activity assays. Synergy was quantified using the Chou–Talalay method, and Combination Index (CI) values were used to determine synergistic interactions. Intracellular ROS levels were measured using the DCFDA assay. Migration and invasion capacities were analyzed using wound healing and Transwell assays. MMP-1, MMP-2, TIMP-1, and TIMP-2 levels were quantified via ELISA with normalization to viable cell counts. Gene expression levels of PI3K/Akt and MAPK/ERK pathway components were measured by qRT-PCR. Bioinformatic analyses (STRING, GeneMANIA, GO, KEGG) were performed to identify common molecular targets and enriched pathways affected by both agents. Results: The combination of sitagliptin and doxorubicin significantly reduced cell viability and demonstrated a synergistic interaction (CI < 1). Combined treatment induced a marked increase in ROS production and significantly elevated apoptosis rates compared to monotherapies. Caspase-8 and caspase-9 activities were also higher in the combination group. Migration and invasion assays revealed substantial suppression of cell motility and invasive capacity. After normalization to viable cell numbers, MMP and TIMP reductions remained significant, confirming true biological inhibition rather than cell-death–related artifacts. qRT-PCR analyses showed downregulation of Akt and ERK expression, indicating suppression of key survival and proliferation pathways. Bioinformatic analyses supported these findings by highlighting enrichment in apoptotic, oxidative stress, and metastasis-related pathways. Conclusions: Sitagliptin enhances the anticancer efficacy of doxorubicin by amplifying ROS-mediated apoptosis, inhibiting migration and invasion, and modulating PI3K/Akt and MAPK/ERK signaling in cervical cancer cells. The combination exhibits a clear synergistic effect and demonstrates strong potential as a supportive therapeutic strategy. These findings warrant further in vivo and clinical-level investigations to evaluate the translational applicability of sitagliptin in cervical cancer therapy. Full article

Background: Chronic inflammation and the development of cancer are closely linked, with components that comprise the tumour microenvironment—including proinflammatory cytokines—exerting essential tumourigenic effects. These proinflammatory cytokines include IL-1β, which has been reported to be overexpressed in several cancers and shown to activate several signalling pathways. These pathways may involve kinases such as AKT (serine/threonine kinase) and Src (Proto-oncogene tyrosine-protein kinase), and have a broad capacity to activate nuclear factors, including NF-κB (Nuclear Factor kappa-light-chain-enhancer of activated B cells), which can regulate the transcription of genes encoding proteins such as cIAP1 (Cellular Inhibitor of Apoptosis Protein 1), Bcl-2 (B-cell lymphoma 2), and cyclin D1, thereby regulating processes like apoptosis and cell cycle inhibition. Objectives: The aim of this study was to investigate the role of IL-1β (Interleukin-1 beta) in regulating cell death and proliferation in RS4:11 leukaemic lymphoblasts via the PI3K (Phosphoinositide 3-kinase)/AKT/Src/NF-κB pathway using an in vitro experimental approach. Methods: We employed flow cytometry to determine the expression levels and phosphorylation status of various proteins; proliferation was assessed using the CCK-8 kit, and apoptosis was evaluated with the Annexin V kit. Results: Our findings indicate that the IL-1β-activated signalling pathway modulates these cellular processes in leukaemic lymphoblasts. Conclusions: We therefore conclude that IL-1β exerts significant effects on cell death and proliferation in leukaemic lymphoblasts through the PI3K/AKT/NF-κB pathway, with the study’s findings indicating that an inflammatory environment may promote such lymphoblasts to acquire neoplastic characteristics. As such, the proteins involved in the effects evaluated in this work could be considered as potential therapeutic targets for the treatment of Acute Lymphoblastic Leukaemia (ALL). Full article

Background/Objectives: Several studies reported that olive leaf extract (OLE) may exert potent anti-cancer activities against human solid and hematological tumors. Such effects are mostly related to the polyphenol oleuropein and its derivatives, which are highly concentrated in OLE. Here, we investigated the anti-tumor effects of OLE in vitro against human acute leukemia and lymphoma cells. Methods: Cell proliferation and apoptosis have been evaluated by flow cytometry (using CFSE and Annexin-V/7AAD, respectively) in the presence or absence of OLE at different concentrations and in combination with or without chemotherapeutic drugs. Cellular pathways have been analyzed using antibody arrays. Results: OLE inhibited cell proliferation and induced apoptosis in B-acute lymphoblastic leukemia (B-ALL) and, to a lesser extent, in lymphomas and acute myeloid leukemia (AML) cell lines. Notably, OLE-induced apoptosis also occurs in primary leukemic blasts from B-ALL patients, both at diagnosis and at relapse, but only marginally in primary AML blasts. The expression and phosphorylation of proteins involved in the induction of apoptosis were modulated by OLE in B-ALL, whereas modest effects were observed in AML. Interestingly, some proteins were modulated in opposite ways in B-ALL and AML, potentially explaining their different responses to OLE. Finally, a synergistic and additive effect was observed for OLE in combination with cytarabine, but not with cyclophosphamide. Conclusions: We may envisage that OLE may be used as a food supplement in B-ALL patients treated with cytarabine, taking advantage of the potentiated effect of chemotherapy, without additional side effects. Full article

The enormous applications of various nanoparticles (NPs) have raised the possibility that humans may be simultaneously exposed to mixtures of them in real life. Realistically, this situation may apply to plastic NPs, mainly derived from the breakdown of larger plastics, and to silver NPs, both of which are among the most frequently detected NPs in the envirnment due to their applications in healthcare, consumer products, and water purification. Although numerous studies have examined the toxicity of plastic and silver NPs individually, knowledge of their combined toxicity remains limited. Hence, the main objective of our study was to investigate the toxicity of high-density polyethene nanoparticles (HDPE NPs), thermally isolated from food-cooking bags, in combination with citrate-stabilised silver nanoparticles (AgNPcit) to Caco-2 and HT29MTX cells growing in 2D monoculture or in 3D triple-culture with Raji cells. Cellular uptake of NPs was quantified from the side-scatter (SSC) signal in flow cytometry; toxicity was evaluated by the neutral red assay; apoptosis was evaluated by the Annexin V method; and induction of oxidative stress was evaluated by a fluorescent method using DCFDA and DHR probes. Both cell lines took up both types of NPs; however, HT29MTX cells were more effective in the NPs’ uptake. Interestingly, HDPE NPs and AgNPcit mutually inhibited each other’s uptake, which suggests a similar mechanism of entry. Both types of NPs were toxic to both cell lines growing in monoculture; Caco-2 cells were more susceptible than HT29MTX. The toxicity was attributed to the induction of oxidative stress and associated apoptosis. In line with the mutual inhibition of the NPs’ uptake, the toxic effect of both NPs in the mixture was less than that expected as the sum of individual treatments. The toxic effects of both NPs or their mixture were less pronounced in the triple-culture Caco-2/HT29MTX/Raji, than in Caco-2 and HT29MTX growing in monoculture. Full article

The rising incidence of cancer has demanded the development of new anti-cancer chemical sources. The presence of phenolics in hazelnut cell cultures has led to the development of new and potential pharmacotherapeutic uses. Hazelnut extract has emerged as a promising candidate due to its high phytochemical content. HCT-116 colorectal cancer IC₅₀ cell viability of Palaz and Tombul hazelnut extracts was determined as 400 μg/mL and 200 μg/mL, respectively. Flow cytometry annexin V-fluorescein isothiocyante (FITC) apoptosis detection indicated apoptosis of Tombul hazelnut extract and Palaz hazelnut extract as 23.53% and 17.47%, respectively. The apoptosis result of flow cytometry was also supported at the protein level. Hazelnut extracts resulted in an increased loss of MMP as well. The loss of MMP has significantly increased from an average of 0.61% to 16.17% in Tombul hazelnut extract and to 20.38% in Palaz hazelnut extract. This is further supported by screening MICU1, MICU2, PPAR-γ, PPARGC1A, UCP1, UCP2, and UCP3 gene expressions. Targeting apoptosis pathways, particularly MMP, is an effective strategy for cancer prevention and treatment. Hazelnut extract contains phenolic compounds, which activate these pathways, resulting in enhanced apoptosis in colorectal cancer cells. The phenolic contents of Palaz and Tombul hazelnut extracts were determined as 271.72 ± 5.3 mg gallic acid equivalent (GAE)/100 g sample dry weight (DW) and 85.23 ± 2.2 mg GAE/100 g sample DW, respectively. Further, hazelnut extract may reduce oxidative stress, contributing to its anti-cancer properties. The extracts could be utilized as functional ingredients in foods and nutraceuticals to assist with cancer prevention and treatment. Full article