Caspase-3 Enzyme Research Articles

Aging Oocytes: Exploring Apoptosis and Its Impact on Embryonic Development in Common Carp (Cyprinus carpio)

Abstract Ovulation, fertilization, and embryo development are orchestrated and synchronized processes essential for the optimal health of offspring. Post-ovulatory aging disrupts this synchronization and impairs oocyte quality. In addition, oocyte aging causes fertilization loss and poor embryo development. This investigation aimed to unravel the endpoint of in vitro oocyte aging in common carp (Cyprinus carpio) to understand the involvement of apoptosis in postovulatory oocyte death. It was observed that the fertilization ability significantly declined (p < 0.001) at 8 hours post-stripping (HPS), subsequently triggering apoptosis in the advanced stage of oocyte aging, i.e. 48 HPS. This process included an increase in proapoptotic transcripts (fas, bax, cathepsin D, caspase 8, caspase 9, and caspase 3a) (p < 0.05), elevated levels of caspase 3 protein (p < 0.05), and activation of caspase 3 enzyme (p < 0.001), a key player in apoptosis, in aging oocytes. Furthermore, the effects of oocyte aging on the embryonic apoptosis machinery were examined in embryos at 5 hours post-fertilization (HPF) and 24 HPF derived from fresh and aged oocytes. Expression of apoptotic genes and caspase enzyme activity remained at the basal level in 5 HPF (early blastula embryos) from both fresh and aged oocytes. In contrast, the zymogenic and active forms of caspase 3 increased in 24 HPF embryos from 8-h-aged oocytes (p < 0.01) compared to those from fresh oocytes. Thus, apoptosis intensified in 24 HPF embryos from aged oocytes without affecting the apoptotic machinery of early blastula embryos. Our findings demonstrate that apoptosis initiated by the Fas/FasL system is an important physiological process accompanying oocyte aging in common carp.

Bolanthus turcicus: a promising antidiabetic with in-vitro antioxidant, enzyme inhibitory and antiadipogenic activities.

It is crucial to investigate new anti-diabetic agents and therapeutic approaches targeting molecules in potential signaling pathways for the treatment of Type 2 diabetes mellitus (T2DM). The objective of the study was to investigate the total phenolic content, antioxidant capacity, α-glucosidase, and α-amylase inhibitory activities of Bolanthus turcicus (B. turcicus), as well as their cytotoxic, anti-adipogenic, anti-diabetic, apoptotic, and anti-migration potential on adipocytes. B. turcicus samples were extracted with methanol (MeOH), ethyl acetate (EA) and aqueous (Aq) solvents. The MeOH extract had the highest phenolic content (81.14mg GAE/g), followed by EA (74.93mg GAE/g) and Aq (51.09mg GAE/g). All extracts exhibited dose-dependent increases in α-glycosidase and α-amylase inhibitory activity. B. turcicus extracts showed cytotoxic effect on adipocytes with IC50 values of MeOH (141.0µg/mL) < Aq (155.3µg/mL) < EA (199.5µg/mL). Furthermore, B. turcicus extracts reduced lipid droplet formation and adipocyte diameter size. All extracts altered cell morphology to resemble fibroblasts. B. turcicus extracts exhibited anti-migratory effect delaying wound healing for up to 96h. The B. turcicus extracts showed a pro-apoptotic effects on adipocytes by increasing Caspase-3 enzyme activity and the population of DAPI-positive cell with apoptotic nuclear-morphology. B. turcicus extracts upregulated the expression of the Glut-4 gene at the mRNA, protein and intracellular level in adipocytes. In conclusion, our findings indicate that B. turcicus not only exhibits strong antioxidant properties and enzyme inhibitory activities but also exerts significant anti-adipogenic and pro-apoptotic effects in adipocytes, thereby providing a comprehensive mechanism through which it may contribute to the management of T2DM. These effects highlight the potential of B. turcicus as a therapeutic agent for improving glucose homeostasis and insulin sensitivity.

Tropisetron-Induced Apoptosis: A Study on SKOV3 Cell Viability and Gene Expression

Background: Ovarian tumors account for 1% of all childhood neoplasms and are the most common genital neoplasm in children. Considering the role of serotonin in the promotion of tumor growth and metastasis in some cancers, tropisetron as a 5-hydroxytryptamine (5-HT3) receptor antagonist can exert anti-neoplastic effects. The current study seeks to determine the association between the use of tropisetron and ovarian cancer­cell lines (SKOV3) by evaluating apoptotic regulators. Materials and Methods: In this experimental study, after the addition of tropisetron to SKOV3 cancer cells at the concentrations of 1, 10, and 100 μM, the MTT assay was employed to assess the cell viability percentage within 24, 48, and 72 hours. The level of expression of Bcl2 and Bax genes after 24 hours were determined by Real Time-PCR, and Caspase 3 enzyme activity was measured by the ELISA method. Differences between groups were statistically analyzed by one-way analysis of variance (ANOVA). A p-value less than 0.05 is considered to be statistically significant. Results: Based on the MTT test, tropisetron significantly decreased the viability of SKOV3 cancer cells. It decreased the levels of Bcl2 expression according to real-time PCR results (P =0.0015) but increased the expression levels of Bax (P = 0.011) in SKOV3 cells. Both caspase-3 enzyme activity and the ratio of Bax to Bcl2 expression (Bax/Bcl2) were increased (P = 0.008 and P = 0.011, respectively). Conclusion: Tropisetron could inhibit tumor cell growth via apoptosis induction and exert proapoptotic effects in an ovarian cancer ¬cell line. Therefore, it seems that it can be considered as a possible chemotherapy drug for ovarian cancer. However, more studies should be conducted in this regard.

Bromelain augments ionizing radiation sensitivity in breast cancer cells: A synergistic approach

BackgroundRadiation-sensitizing agents, such as Bromelain (pineapple extract) have been shown to significantly impact cancer cell cycle arrest. This study investigates the anticancer properties of Bromelain on MCF-7 breast cancer cell line under both two-dimensional (2D) and three-dimensional (3D) culture conditions. MethodsBromelain enzyme was initially applied at concentrations of 75, 96, and 115 μg/ml to sensitize the MCF-7 cells under both 2D and 3D culture conditions. Following treatment, the cells were irradiated with doses of 2, 4, and 6 Gy. The synergistic effects of bromelain enzyme treatment and gamma irradiation on MCF-7 cells were assessed using the MTT assay, neutral red uptake assay, alkaline comet assay, nitric oxide (NO) release measurement, catalase activity assay, reduced glutathione quantification, and caspase 3 assay. Data were analyzed and presented as Mean ± SD using GraphPad InStat software V. 8. ResultsThe MTT assay indicated that the combination of bromelain and ionizing radiation have synergistic effects compared to cells treated with 2 Gy of X-ray alone. The results from of the MTT assay indicated showed that 75 μg/ml of bromelain did not induce exhibit significant cytotoxicity. Consistent with the MTT assay results, the Neutral red uptake assay demonstrated increased mortality in MCF-7 cells following treatment with both bromelain and ionizing radiation. Additionally, treatment with bromelain and ionizing radiation resulted in elevated levels of released NO, GSH content, and catalase enzyme activity compared to cells treated with bromelain alone. Furthermore, the Combination of ionizing radiation and bromelain synergistically enhanced caspase-3 enzyme activity and increased the number of necrotic and apoptotic cells compared to treatment with bromelain alone. ConclusionBromelain's capacity to induce apoptosis and enhance radio-sensitization offers promising potentialfor improving cancer treatment outcomes. By promoting programmed cell death and increasing the sensitivity of cancer cells to radiation, bromelain could contribute to more effective treatment regimens while potentially reducing the adverse effects associated with more aggressive therapies.

Mechanisms of ROS-induced mitochondria-dependent apoptosis in Phaeodactylum tricornutum under AgNPs exposure

IntroductionNanomaterials such as silver nanoparticles (AgNPs) have gained widespread application across various fields. However, the large-scale production and application of AgNPs have raised concerns about their distribution in the environment and potential pollution issues.MethodsThis study investigates the toxic effects of AgNPs on the diatom Phaeodactylum tricornutum by employing electron microscopy for cellular observation, quantifying apoptotic cell numbers, and measuring antioxidant indicators. The research examines how varying concentrations of AgNPs induce stress in P. tricornutum and the specific mechanisms of the toxic effect.ResultsThe findings reveal that AgNPs induce apoptosis in P. tricornutum cells by triggering a mitochondria-mediated pathway, marked by a decrease in mitochondrial membrane potential and the activation of caspase enzymes. Additionally, AgNP exposure results in an overproduction of reactive oxygen species (ROS) within the algal cells, leading to lipid peroxidation of the cell membrane and a consequent increase in malondialdehyde (MDA) levels. This oxidative stress response induces the upregulation of antioxidant enzyme activities in an attempt to mitigate the excessive ROS.DiscussionROS is identified as the primary factor responsible for inducing mitochondria-mediated apoptosis. The research results will provide a theoretical basis for understanding the toxic effects and mechanisms of AgNPs on marine microalgae.

Toxic Effects of Synthesized Bismuth Oxide/Reduced Graphene Oxide (Bi2O3/RGO) Nanocomposites in Two Distinct Mammalian Cell Lines: Role Oxidative Stress and Apoptosis.

Researchers have shown substantial interest in bismuth oxide/reduced graphene oxide (Bi2O3/RGO) nanocomposites due to their superior features that are not achievable by each material alone. The growing applications and manufacturing of Bi2O3/RGO nanocomposites have raised concerns regarding their potential human health risks. This work was designed to explore the possible toxicity mechanisms of Bi2O3/RGO nanocomposites in two distinct mammalian cell lines, normal rat kidney cells (NRK52E) and human liver cancer cells (HepG2). Bi2O3/RGO nanocomposites were prepared by a simple hydrothermal technique. X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and dynamic light scattering (DLS) were used to characterize the synthesized nanocomposites. The cytotoxicity of Bi2O3/RGO nanocomposites in NRK52E and HepG2 cells was examined by MTT cell viability assay. Reactive oxygen species (ROS) and glutathione (GSH) were measured as the biomarkers of oxidative stress. The apoptosis study was carried out by measuring several parameters, including cell cycle and caspase-3. High-quality Bi2O3/RGO nanocomposites of ≈33-38 nm size without impurities, where crystalline Bi2O3 particles are evenly attached to the RGO sheets. Bi2O3/RGO nanocomposites exhibit cytotoxic effects on NRK52E and HepG2 cells, which were dose- and time-dependent. Interestingly, NRK52E exhibited marginally higher vulnerability to Bi2O3/RGO nanocomposites compared to HepG2. Bi2O3/RGO nanocomposites also cause a dose-dependent increase in ROS production and a decrease in GSH levels. Exposing NRK52E and HepG2 cells to Bi2O3/RGO nanocomposites results in activation of the caspase-3 enzyme and chromosomal condensation. The apoptotic response of Bi2O3/RGO nanocomposites against both types of cells was further confirmed by AO-EB dual staining and altered cell cycle. This study demonstrated that the toxicity of Bi2O3/RGO nanocomposites in both NRK52E and HepG2 cells is attributed to their ability to produce ROS, leading to apoptosis and cell cycle arrest as a consequence of oxidative stress.

Cytotoxic effect of polystyrene nanoplastics in human umbilical vein endothelial cells (HUVECs) and normal rat kidney cells (NRK52E)

Plastics play a crucial role in nearly each aspect of societal production and everyday life, therefore making them one of the most widespread pollutants on a global scale. Because synthetic plastics are not entirely biodegradable, they often remain in the environment and fragment into micro- and nanoplastic particles. The detrimental impacts of nanoplastics on the environment and human health have received substantial worldwide interest in recent years. However, the effects of nanoplastics on human health have not yet been fully explored. Our study aimed to investigate the cytotoxic effects of polystyrene nanoplastics on two distinct mammalian cell lines: normal rat kidney cells (NRK52E) and human umbilical vein endothelial cells (HUVECs). Results showed that polystyrene nanoplastics generate cytotoxicity in both NRK52E and HUVECs in a concentration- and time-dependent manner. Additionally, polystyrene nanoplastics induced pro-oxidant levels (e.g., reactive oxygen species and hydrogen peroxide) and reduced antioxidants (glutathione content and glutathione peroxidase enzyme activity) in both types of cells. We also found that polystyrene nanoplastics cause apoptosis in both NRK52E and HUVECs, as shown by the activation of the caspase-3 enzyme and the loss of mitochondrial membrane potential. Interestingly, it was noticed that the vulnerability of HUVECs cells against polystyrene nanoplastics was a little higher than that of NRK52E cells. Also, the cell toxicity caused by polystyrene nanoplastics in NRK52E and HUVECs was effectively alleviated by co-exposure to a reactive oxygen species inhibitor, N-acetyl-cysteine. This suggests that oxidative stress could be one of the possible pathways by which polystyrene nanoplastics cause cell toxicity. The present work warrants future study to explore the toxicity mechanisms of polystyrene nanoplastics in appropriate in vivo models.

Pyroptosis and sepsis-associated acute kidney injury.

Pyroptosis is a form of programmed cell death triggered by inflammatory caspases, dependent on the gasdermin (GSDM) family proteins forming membrane pores in the plasma membrane, with GSDM proteins serving as the executors of pyroptosis. This process can activate a robust inflammatory response through a cascade effect. Sepsis-associated acute kidney injury (SA-AKI) is a classical inflammatory disease with no specific therapeutic drug available. Studies have highlighted the role of pyroptosis in the onset and progression of SA-AKI, yet the specific renal cell populations affected by pyroptosis and the detailed regulatory mechanisms remain unclear. Pyroptosis may be closely related to SA-AKI, with current strategies for regulating pyroptosis focusing on targeting inflammasomes, key caspase enzymes, GSDM proteins, and downstream inflammatory factors. Although these strategies still present some off-target effects or side effects, they provide a foundation for research into sepsis-targeted therapies and clarify future research directions and the necessity of such studies.

Hepatoprotective effect of date fruit extract against ethanol-induced apoptosis in human hepatoma (HepG2) cells

Ethanol is a well-known hepatotoxic agent and date fruits have been associated with their biological actions. In current study, we have investigated the hepatoprotective potential of DFE on ethanol-induced cellular damages in human hepatoma (HepG2) cells. The hepatoprotective potential was assessed by exposing the HepG2 cells to non-toxic concentrations (15, 30, and 60 μg/mL) of DFE for 24 h; then toxic concentration (500 μM) of ethanol. Our results demonstrated that pretreatment with DFE significantly prohibited ethanol-induced hepatotoxicity in HepG2 cells. We observed that DFE treatment increased cell viability, reduced LDH leakage, restored cellular morphology, and inhibited caspase-3 enzyme activity in a dose dependent way, induced by ethanol. Further DFE was also effective in restoring the LPO, GSH, and catalase levels towards normal altered by ethanol. Our results also revealed that ethanol-induced ROS generation was significantly inhibited by DFE. The ethanol-induced mRNA expression of apoptotic related genes (p53, caspase-3, caspase-7, Bax, and Bcl-2) were also normalized by pretreatment with DFE. The findings from this study indicated that DFE can significantly protect HepG2 cells against ethanol-induced hepatotoxicity. Our study also provides scientific validation for the traditional use of DFE, aiming to understand its hepatoprotective potential. Altogether, to the best of our knowledge, this is the first study demonstrated that ethanol-induced hepatotoxicity can be prohibited by the DFE. Thus, DFE has a potential application in nutraceuticals as a therapeutic agent to prevent liver diseases.

Differential effects of montelukast and zafirlukast on MDA‑MB‑231 triple‑negative breast cancer cells: Cell cycle regulation, apoptosis, autophagy, DNA damage and endoplasmic reticulum stress.

Montelukast and zafirlukast, cysteinyl leukotriene receptor antagonists (LTRAs), trigger apoptosis and inhibit cell proliferation of triple‑negative breast cancer MDA‑MB‑231 cells. By contrast, only zafirlukast induces G0/G1 cell cycle arrest. The present study compared the effects of these drugs on proteins regulating cell proliferation, apoptosis, autophagy, and endoplasmic reticulum (ER) and oxidative stress using reverse transcription‑quantitative PCR, western blotting and flow cytometry. The expression of proliferating markers, Ki‑67 and proliferating cell nuclear antigen, was decreased by both drugs. Zafirlukast, but not montelukast, decreased the expression of cyclin D1 and CDK4, disrupting progression from G1 to S phase. Zafirlukast also increased the expression of p27, a cell cycle inhibitor. Both drugs decreased the expression of anti‑apoptotic protein Bcl‑2 and ERK1/2 phosphorylation, and increased levels of the autophagy marker LC3‑II and DNA damage markers, including cleaved PARP‑1, phosphorylated (p)‑ATM and p‑histone H2AX. The number of caspase 3/7‑positive cells was greater in montelukast‑treated cells compared with zafirlukast‑treated cells. Montelukast induced higher levels of the ER stress marker CHOP compared with zafirlukast. Montelukast activated PERK, activating transcription factor 6 (ATF6) and inositol‑requiring enzyme type 1 (IRE1) pathways, while zafirlukast only stimulated ATF6 and IRE1 pathways. GSK2606414, a PERK inhibitor, decreased apoptosis mediated by montelukast, but did not affect zafirlukast‑induced cell death. The knockdown of CHOP by small interfering RNA reduced apoptosis triggered by montelukast and zafirlukast. In conclusion, the effects on cell cycle regulator proteins may contribute to cell cycle arrest caused by zafirlukast. The greater apoptotic effects of montelukast may be caused by the higher levels of activated caspase enzymes and the activation of three pathways of ER stress: PERK, ATF6, and IRE1.

Novel Thiazole-Hydrazide Derivatives and Their Anticancer Properties

Objective: Cancer is described as uncontrolled cell division, and it is a major problem in Türkiye, as well as around the world. Current treatment options are insufficient in some cases, particularly the treatment rate for lung cancer cases, which is very low. Meanwhile, current pharmaceuticals have several side effects, such as drug-drug interactions, and cognitive disorders. Additionally, developing drug resistance is a major problem for current and future management of the disease. Accordingly, the search for new molecules or alternative treatment options is actively achieved. Methods: In this study, eight novel thiazole-hydrazide analogs were designed and synthesized, and their structural elucidation was performed via HRMS, 1H-NMR, and 13C-NMR. Their biological activity profile was investigated on A549 lung carcinoma and MCF7 breast adenocarcinoma cells. To determine the selective cytotoxicity on cancer cells, they were also tested against NIH/3T3 healthy cell line. Besides that, an in silico study was performed to understand the binding modes of the compounds. Results: The results showed that in the serial 4f and 4g, the most bulky analogues, showed no inhibition against any cell type, even at the highest concentration tested. On the other hand, 4a, 4b, 4d, 4e, and 4h showed less cytotoxicity on healthy cells than A549 cells, so they exhibited significant cytotoxicity and a selective profile against A549 cancer cells. While they also inhibited MCF7 cells. The major point is that para-chlorophenyl analogs at the fourth position on thiazole (4a and 4d) displayed a better anticancer profile than ortho-chlorophenyl analogs. These two compounds were also investigated for their apoptotic effects using in silico studies. Both experimental and in silicon studies revealed that the combination of thiazole and hydrazinoacetyl has a significant impact against cancer cells, and in silico study also suggested that tri-substitute thiazole ring has anticancer potential that induced cancer cell death via apoptosis. Conclusion: Results of this study was presented that compound 4a was the most potent compound against lung cancer cells (A549) and 4d was the most potent compound against breast cancer cells (MCF-7). Furthermore, analyzing the molecular docking study for promising compounds (4a and 4d) suggested that interactions with the loop region residues have a pivotal role in inducing caspase-3 enzyme activity. It was concluded that hybridization of thiazole and hydrazinoacetyl moieties is responsible for the anticancer activity.

High levels of mitochondrial dynamics, autophagy, and apoptosis contribute to stable testicular status in hibernating Daurian ground squirrels

Daurian ground squirrels (Spermophilus dauricus) experience various stress states during winter hibernation, but the impact on testicular function remains unclear. This study focused on the effects of changes in testicular autophagy, apoptosis, and mitochondrial homeostasis signaling pathways at various stages on the testes of Daurian ground squirrels. Results indicated that: (1) During winter hibernation, there was a significant increase in seminiferous tubule diameter and seminiferous epithelium thickness compared to summer. Spermatogonia number and testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels were higher during inter-bout arousal, suggesting that the testes remained stable during hibernation. (2) An increased number of mitochondria with intact morphology were observed during hibernation, indicating that mitochondrial homeostasis may contribute to testicular stability. (3) DNA fragmentation was evident in the testes during the hibernation and inter-bout arousal stages, with the highest level of caspase3 enzyme activity detected during inter-bout arousal, together with elevated levels of Bax/Bcl-2 and Lc3 II/Lc3 I, indicating an up-regulation of apoptosis and autophagy signaling pathways during hibernation. (4) The abundance of DRP1, MFF, OPA1, and MFN2 proteins was increased, suggesting an up-regulation of mitochondrial dynamics-related pathways. Overall, testicular autophagy, apoptosis, and mitochondrial homeostasis-related signaling pathways were notably active in the extreme winter environment. The well-maintained mitochondrial morphology may favor the production of reproductive hormones and support stable testicular morphology.

Exogenous application of melatonin attenuates waterlogging stress through adopting quiescence adaptation technique in tomato seedlings

Waterlogging (WL) is a major limiting factor in global crop production and seriously limits growth and yield improvement in low-lying rainfed regions. Melatonin (MT) is a vital phytohormone that functions as a "master regulator" in multiple facets related to plant growth and development, in addition to maintaining a potential role in response to stresses. However, the pharmacological role of MT in attenuating waterlogging stress in tomato largely remains elucidated. The objective of the current investigation is to justify the physiological regulatory mechanism of tomato seedlings exposed to WL and the putative functions of MT to mitigate the adverse effects of WL. Tomato seedlings were grown on substrate (peat: vermiculite, 2:1, v/v) and at the 4th leaf stage subjected to waterlogging stress for 10 days and seedlings were foliar sprayed with melatonin during waterlogging stress. The results revealed that WL significantly arrested tomato seedlings growth, and reduced pigment content coincided with enhanced leaf senescence. MT supplementation attenuated WL-induced oxidative damage through increasing osmoprotectants activity, elevating antioxidant enzyme functioning synchronized with inhibiting excess reactive oxygen species (ROS) production. The concentrations of MT (28 %), abscisic acid (ABA, 170 %), and 1-aminocyclopropane-1-carboxylic acid (ACC, 129 %) were increased, while indole acetic acid (IAA, 15 %), jasmonic acid (JA, 55 %) and gibberellic acid (GA3, 26 %) content were decreased in only WL seedlings roots relative to control seedlings. The core anaerobic respiration enzyme alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC) activity were elevated by 127 % and 163 %, respectively at day 10 in WL+MT received seedlings than control. WL treatment varyingly contributed on nutrient content, as evidenced that N+, K+, and Ca2+ content decreased, whereas Mn2+, Fe2+, and Mg2+ content increased and MT addition reversed their concentrations under similar stress conditions. Exogenous MT promoted WL-tolerance of tomato by positively suppressing respiratory burst oxidase homologs (RBOH)-regulating gene expression while up-regulating ethylene biosynthesis gene transcription. Most importantly, programmed cell death (PCD) regulated enzyme caspase-3 activity concurred with PCD-induced gene (caspase-3, pirin, TBN1) expression significantly inhibited by MT application. In general, these findings reveal that external supplementation with MT can improve plant tolerance to WL through complex processes and multifaceted mechanisms.

G protein-coupled estrogen receptor agonist G-1 decreases ADAM10 levels and NLRP3-inflammasome component activation in response to Staphylococcus aureus alpha-hemolysin.

The G protein-coupled estrogen receptor, also known as GPER1 or originally GPR30, is found in various tissues, indicating its diverse functions. It is typically present in immune cells, suggesting its role in regulating immune responses to infectious diseases. Our previous studies have shown that G-1, a selective GPER agonist, can limit the pathogenesis mediated by Staphylococcus aureus alpha-hemolysin (Hla). It aids in clearing bacteria in a mouse skin infection model and restricts the surface display of the Hla receptor, ADAM10 (a disintegrin and metalloprotease 10) in HaCaT keratinocytes. In this report, we delve into the modulation of GPER in human immune cells in relation to the NLRP3 inflammasome. We used macrophage-like differentiated THP-1 cells for our study. We found that treating these cells with G-1 reduces ATP release, decreases the activity of the caspase-1 enzyme, and lessens cell death following Hla intoxication. This is likely due to the reduced levels of ADAM10 and NLRP3 proteins, as well as the decreased display of the ADAM10 receptor in the G-1-treated THP-1 cells. Our studies, along with our previous work, suggest the potential therapeutic use of G-1 in reducing Hla susceptibility in humans. This highlights the importance of GPER in immune regulation and its potential as a therapeutic target.

Anticancer activity of zinc-tin oxide/dextran/geraniol nanocomposites against 1,2-dimethylhydrazine-induced colon cancer in rats: In vitro and In vivo study

BackgroundColorectal cancer (CRC) is the third most common cause of cancer-related mortality and the fourth most common cancer worldwide. Nanotechnology has experienced various developments, resulting in the emergence of novel cancer treatment methods. ObjectiveThe current work was planned to synthesize and characterize the zinc-tin-oxide/dextran/geraniol nanocomposites (ZSD-Ger-NCs) and evaluate their anticancer properties against colon cancer in vitro and in vivo studies. MethodologyThe ZSD-Ger-NCs were synthesized by the co-precipitation method and characterized using several techniques. The in vitro studies were conducted to evaluate the effect of ZSD-Ger-NCs on growth, apoptosis, and caspase activities in the HCT-116 cells. In vivo experiments were performed on 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in rats. The effect of ZSD-Ger-NCs on body weight, and tumor volume was studied. The oxidative biomarker levels and myeloperoxidase (MPO) activity were analyzed using the corresponding kits. The colon tissues were subjected to histopathological analysis and immunohistochemical analysis. ResultsThe findings of various characterization methods revealed the generation of agglomerated ZSD-Ger-NCs with a well-defined shape and an average size of 197.40 nm. The ZSD-Ger-NCs treatment inhibited growth and promoted apoptosis in HCT-116 cells via augmenting caspase enzymes. In DMH-induced rats, the ZSD-Ger-NCs effectively increased body weight and reduced tumor volume. The ZSD-Ger-NCs treatment effectively mitigated the oxidative stress in the DMH-induced rats by augmenting the antioxidants. Histopathological findings proved that the ZSD-Ger-NCs reduced epithelial damage and dysplasia in the colon tissues. The expression of COX-2 was down-regulated by the ZSD-Ger-NCs treatment in DMH-induced rats. ConclusionThe findings of this work proved the anticancer properties of synthesized ZSD-Ger-NCs in inhibiting DMH-induced colon cancer development in rats. Therefore, it can be a talented anticancer candidate for treating colon cancer.

Molecular probes for monitoring pyroptosis: design, imaging and theranostic application.

Pyroptosis is a recently discovered process of programmed cell death that is linked with tumor progression and potential treatment strategies. Unlike other forms of programmed cell death, such as apoptosis or necrosis, pyroptosis is associated with pore-forming proteins gasdermin D (GSDMD), which are cleaved by caspase enzymes to form oligomers. These oligomers are then inserted into the cell surface membrane, causing pores to consequently result in rapid cell death. Pyroptosis, in conjunction with immunotherapy, represents a promising avenue for prognostication and antitumor therapy, providing a more precise direction for disease treatment. To gain deeper insight into the mechanisms underlying pyroptosis in real-time, non-invasive and live cell imaging techniques are urgently needed. Non-invasive imaging techniques can enhance future diagnostic and therapeutic approaches for inflammatory diseases, including different types of tumors. This review article discusses various non-invasive molecular probes for detecting pyroptosis, including genetic reporters and nanomaterials. These strategies can enhance scientists' understanding of pyroptosis and help discover personalized and effective ways to treat inflammatory diseases, particularly tumors.

Determination of the therapeutic effects of apigenin on chronic myeloid leukemia stem cells; a mechanistic approach

Aims: Chronic myeloid leukemia (CML) is a type of leukemia characterized by the Philadelphia (Ph) chromosome encoding the BCR-ABL protein. Significant success has been achieved in the treatment of CML with the development of agents that target this protein and inhibit its activity, but the resistance that develops in patients against these drugs limits absolute success. One of the main reasons for the resistance problem is the ineffectiveness of current therapies for CML stem cells. Apigenin (4',5,7- trihydroxyflavone) is a flavanoid found in various vegetables and fruits and has the effects of suppressing proliferation and inducing apoptosis in different cancer types. In this study, we aimed to determine the therapeutic potential of apigenin on K562 CML stem cells. Methods: The effects of apigenin on the proliferation of K562 CML stem cells were determined by an XTT cell proliferation assay. Apoptotic effects of apigenin on K562 CML stem cells were determined by changes in mitochondrial membrane potential (MZP), caspase-3 enzyme activity, and the Annexin-V method using flow cytometry. Results: The proliferation of K562 CML stem cells exposed to increasing doses of Apigenin (1-40 nM) for 48 hours decreased dose-dependently, and the IC50 value of Apigenin was calculated at 3 nM. Compared to the control group, an increase in caspase-3 enzyme activity was calculated in stem cells treated with apigenin at the same doses. Disruptions in the mitochondrial membrane potential of Apigenin-treated CML stem cells and Annexin-V assay results also showed that Apigenin dose-dependently induced apoptosis and caused significant increases in the apoptotic cell population. Conclusion: It was shown for the first time in this study that apigenin may have therapeutic effects on CML stem cells. It was determined that this effect of apigenin was achieved by triggering caspase-3 enzyme activity and disruptions in the mitochondrial membrane.

H-ras-targeted genetic therapy remarkably surpassed docetaxel treatment in inhibiting chemically induced hepatic tumors in rats

AimsHepatocellular carcinoma (HCC) is still a leading cause of cancer-related death worldwide. But its chemotherapeutic options are far from expectation. We here compared H-ras targeted genetic therapy to a commercial docetaxel formulation (DXT) in inhibiting HCC in rats. Main methodsAfter the physicochemical characterization of phosphorothioate-antisense oligomer (PS-ASO) against H-ras mutated gene, the PS-ASO-mediated in vitro hemolysis, in vivo hepatic uptake, its pharmacokinetic profile, tissue distribution in some highly perfused organs, its effect in normal rats, antineoplastic efficacy in carcinogen-induced HCC in rats were evaluated and compared against DXT treatment. Mutated H-ras expression by in situ hybridization, hep-par-I, CK-7, CD-15, p53 expression patterns by immunohistochemical methods, scanning electron microscopic evaluation of hepatic architecture, various hepatic marker enzyme levels and caspase-3/9 apoptotic enzyme activities were also carried out in the experimental rats. Key findingsPS-ASO showed low in vitro hemolysis (<3 %), and had a sustained PS-ASO blood residence time in vivo compared to DTX, with a time-dependent hepatic uptake. It showed no toxic manifestations in normal rats. PS-ASO distribution was although initially less in the lung than liver and kidney, but at 8 h it accumulated more in lung than kidney. Antineoplastic potential of PS-ASO (treated for 6 weeks) excelled in inhibiting chemically induced tumorigenesis compared to DTX in rats, by inhibiting H-ras gene expression, some immonohistochemical modulations, and inducing caspase-3/9–mediated apoptosis. It prevented HCC-mediated lung metastatic tumor in the experimental rats. SignificancePS-ASO genetic therapy showed potential to inhibit HCC far more effectively than DXT in rats.

The Effect of Continuous Positive Airway Pressure (CPAP) Therapy on Serum Caspase-3 Level in Patients with Obstructive Sleep Apnea (OSA).

Intermittent hypoxemia has an important role in the physiopathogenesis of obstructive sleep apnea (OSA) complications. Increased apoptosis due to intermittent hypoxemia may be an important clinical entity in OSA. In this study, we aimed to evaluate caspase-3 enzyme level, which is an indirect marker of increased apoptosis in patients with OSA and to evaluate the effect of OSA treatment with continuous positive airway pressure on caspase-3 enzyme level. This study included 141 consecutive patients admitted to the sleep-disordered breathing laboratory within 6 months. Caspase-3 was measured in routine blood samples obtained on the morning of polysomnography (PSG) performed at night. The compliance of the patients to CPAP treatment was evaluated and caspase-3 levels were checked again after treatment. A total of 141 patients, 39 females (27,7%) and 102 males (72,3%) were included in the study. The mean age of the patients was 49 ± 12 years (min-17, max-77). According to PSG results, OSA was detected in 95.7% (135/141) of the cases. Mild OSA was 35 (24.8%), moderate OSA 39 (27.7%) and severe OSA 61 (43.3%) cases. Median caspase-3 enzyme levels were similar in men and women in the study group. There was no statistically significant difference in hemogram parameters and caspase-3 enzyme levels between the groups divided according to the presence and severity of OSA. It was determined that caspase-3 enzyme level did not change significantly after 3 months of CPAP treatment in OSA compared to pretreatment. Caspase-3 was found to have a negative correlation with both the percentage of daily use of CPAP therapy and the percentage of CPAP device use for more than 1 h per night. It was found that the control caspase-3 level decreased statistically significantly as the percentage of daily use of CPAP therapy increased (r = -0.397, p = 0.030). It was found that the control caspase-3 level decreased statistically significantly as the percentage of CPAP therapy use for more than 1 h per night increased (r = -0.411, p = 0.024). The results of this study did not reveal a relationship between the severity of OSA and caspase-3 levels. However, blood caspase-3 levels decreased as treatment compliance increased, suggesting that CPAP treatment may correct increased apoptosis in OSA. There is a need for more comprehensive studies on this issue.

Abstract 4700: Induced phenotype targeted ADC by using caspase enzymatic cleavable peptide linker to overcome tumor heterogeneity

Abstract Antibody-drug conjugates (ADCs) have ushered in a new era of precision cancer therapy, and the caspase-3-cleavable linker has demonstrated its potential as a game-changing addition to the ADC toolkit. The DEVD peptide, specifically cleaved by caspase and lysosomal enzymes, in combination with the KG peptide, provides the necessary flexibility to achieve targeted cancer cell apoptosis. This research explores the key attributes and actions of the linker, aiming to highlight its potential for revolutionizing ADC therapy. The caspase-3-cleavable linker operates through caspase-3 activation during apoptosis. After targeting cancer cells with specific antigens, the ADC is internalized into the cells, and the linker is cleaved by caspase-3, releasing the drug and initiating apoptosis. The specificity of caspase-3 extends the drug’s effect to neighboring cells, even those lacking the target antigen. This cascade effect ensures the continuous activation of ADCs until all cancer cells are eradicated. Firstly, it overcomes tumor heterogeneity by activating the drug outside of the cell. Traditional bystander killing effects rely on internalization followed by drug release and penetration of neighbor cancer cells. However, this linker's unique mechanism ensures that the drug is activated by caspase-3 outside the cell without the need for internalization, effectively targeting all cancer cells within the tumor tissue. This is achieved through the induced phenotype targeting effect (IPTE), which involves caspase-3 cleavage of the linker, inducing a phenotype in the tumor tissue that targets all cancer cells, regardless of their specific markers. Secondly, it maintains therapeutic efficacy even as the target antigen decreases. Current targeted therapies face a challenge as target antigens may diminish during treatment, leading to reduced drug effectiveness. However, the unique mechanism of the linker ensures that the drug continues to be activated as long as there is caspase-3 secretion, resulting in continuous activation of ADCs in the tumor, with a feedback amplification loop that ensures therapeutic efficacy until all cancer cells are eliminated. Lastly, the linker's superior physicochemical properties allow for a maximum drug-to-antibody ratio (DAR). Its hydrophilic nature enables independent drug conjugation, increasing therapeutic potential and mitigating aggregation issues. This innovation addresses the current limitations in ADC conjugation technologies. In summary, the caspase-3-cleavable linker represents a significant leap forward in the field of ADCs. Its unique attributes offer promising prospects for more effective and targeted cancer treatments. This linker is poised to revolutionize the landscape of targeted cancer therapy, bringing us one step closer to the dream of eradicating cancer cells with precision and efficiency. Citation Format: Ha Kyeong Lee, Byoungmo Kim, Hyo Won Chang, Sang Yoon Kim, Youngro Byun. Induced phenotype targeted ADC by using caspase enzymatic cleavable peptide linker to overcome tumor heterogeneity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4700.