Apoptotic Proteins Caspase-3 Research Articles

Discovery of New Pyrazole-Tosylamide Derivatives as Apoptosis Inducers Through BCL-2 Inhibition and Caspase-3 Activation.

In this presented study, a series of new carbonitrile-substituted pyrazole-tosyl amide derivatives were designed and synthesized according to previous studies. The antiproliferative effects of the synthesized compounds on MDA-MB-231, MCF-7, HepG2, PC-3, and A549 cancer cell lines were assessed by MTT assay compared with non-cancerous cells. The results demonstrate that compounds 9d, 9e, and 9f had a higher antiproliferative effect (IC50 <10 μM) against both breast cancer cells. To investigate the ability of these compounds (9d-f) to induce apoptosis against breast cancer cells, BCL-2 levels and Caspase-3 activities of compound-treated breast cancer cell lines were measured by ELISA. The results revealed that these compounds significantly inhibited the levels of anti-apoptotic protein BCL-2 and increased the activity of apoptotic protein Caspase-3 in MDA-MB-231 and MCF-7 cells. Molecular docking studies confirmed that the selected compounds have high binding affinity towards the active site in the crystal structures of BCL-2 and Caspase-3. Moreover, drug-likeness and pre-ADMET evaluation showed that the compounds had suitable drug properties. This study may be a new milestone in terms of the promising importance of carbonitrile-substituted pyrazole-tosyl amide scaffolds as apoptosis-inducing agents for cancer therapy in the future.

Evaluation of Vincamine Loaded with Silver Nanoparticles as a New Potential Therapeutic Agent Against Ehrlich's Solid Carcinoma in Mice.

Vincamine, a monoterpenoid indole alkaloid with vasodilatory properties, is extracted from the leaves of Vinca minor. The present study aimed to determine the potential anticancer effects of vincamine loaded in silver nanoparticles (VCN-AgNPs) in mice with Ehrlich solid carcinoma (ESC). After tumor transplantation, the mice were divided into five groups: ESC, ESC+Cisplatin (CPN; 5 mg/kg), ESC+VCN (40 mg/kg), ESC+AgNPs (6 mg/kg), and ESC+VCN-AgNPs (20 mg/kg). The administration of VCN-AgNPs to ESC-bearing mice improved their survival rate and reduced their body weight, tumor size, and tumor weight compared to the ESC group. Furthermore, VCN-AgNPs intensified oxidative stress in tumor tissues, as evidenced by elevated levels of lipid peroxidation (LPO) and nitric oxide (NO), along with a reduction in the levels of the antioxidants investigated (GSH, GPx, GR, SOD, CAT, and TAC). Furthermore, VCN-AgNPs increased the apoptotic proteins Bax and caspase-3, decreased the anti-apoptotic protein (Bcl-2), increased the inflammatory markers TNF-α and IL-1β, and inhibited angiogenesis by lowering VEGF levels in tumor tissues, all of which led to apoptosis. Furthermore, histopathological studies showed that VCN-AgNPs suppressed the progression of Ehrlich carcinoma and induced the formation of clusters of necrotic and fragmented tumor cells. VCN-AgNPs possess cytotoxic and genotoxic effects against ESC because of their pro-oxidant, pro-apoptotic, pro-inflammatory, and antiangiogenic effects. Additionally, the combination of VCN-AgNPs was more effective and safer than chemically synthesized AgNPs, as indicated by an increase in the lifespan of animals and the total tumor inhibition index.

10-Hydroxy Decanoic Acid and Zinc Oxide Nanoparticles Retrieve Nrf2/HO-1 and Caspase-3/Bax/Bcl-2 Signaling in Lead-Induced Testicular Toxicity.

There has been a significant increase in human exposure to heavy metals (HMs) over the course of the previous century, primarily due to the extensive industrial processes. Male infertility is a prominent complication associated with lead exposure, wherein lead has the potential to accumulate within the testes, resulting in oxidative stress and inflammation. In addition, 10-hydroxydecanoic acid (10-HDA) is a component found in the secretions of worker bees and possesses the capacity to mitigate oxidative stress and prevent inflammation. Due to their advantageous properties, zinc oxide nanoparticles (ZnO-NPs) possess a wide range of applications in the field of biomedicine. This study aimed to assess the therapeutic effect of 10-HDA and ZnO-NPs on testicular toxicity in rats induced by lead acetate (PbAc). PbAc was administered orally for a period of 3months. Following that, 10-HDA and/or ZnO-NPs were administrated for 1month. PbAc deformed seminal analysis, decreased seminal fructose and sex hormonal levels, and resulted inthe development of histopathological complications. Additionally, PbAc increased MDA and decreased Nrf2 and HO-1 expression, confirmed by the declined antioxidant defense system. Furthermore, an increase in testicular inflammatory markers and the Bax/Bcl-2 ratio was observed subsequent to the administration of PbAc. The administration of 10-HDA and ZnO-NPs demonstrated significant efficacy in the restoration of semen quality, pituitary/gonadal hormones, antioxidants, and testicularhistoarchitecture. Moreover, 10-HDA and ZnO-NPs decreased testicular inflammatory markers and apoptotic proteins (caspase-3 and Bax expression levels). In conclusion, combining 10-HDA and ZnO-NPs demonstrated synergistic potential in treating PbAc-induced testicular toxicity, thereby presenting a promising approach in nanomedicine and natural drugs.

Abstract C026: Analyzing the effect of annonacin on diverse breast cancer cell lines as a potential therapeutic agent for triple negative breast cancer

Abstract Triple negative breast cancer (TNBC) grows and spreads faster than most invasive breast cancer. Additionally, available treatment options and prognosis are poor. Due to limited available treatment options, finding the appropriate and effective chemotherapeutic agents for TNBC patients is a key research effort. Importantly, differential therapeutic response reported between ethnic and racial groups has prompted a need for better treatment modalities. In previous studies, Annonacin (Annona Muricata-a plant with anti-inflammatory properties) has been tested and shown to have cytotoxic effects on various cancer cell lines. The objective of this study is to analyze the effects of Annonacin across racial backgrounds. For this study, we used two TNBC cell lines; MDA-MB-468 (derived from a 51-year old African American female) and MDA-MB-231 (derived from a 40-year old Caucasian American female). Our hypothesis is that Annonacin will inhibit cell proliferation in both TNBC cell lines, potentially giving rise to a beneficial therapeutic agent that will address health disparity. To assess the efficacy of Annonacin, cell viability was measured using a MTT assay providing IC50 values for each cell line. The 50% inhibitory concentrations were 8.5 uM and 15 uM for MDA-MB-468 and MDA-MB-231, respectively. Cell cycle analysis was performed using flow cytometry to assess the effect of Annonacin on the induction of cell cycle arrest. MDA-MB-231 was arrested at the G2 phase in a dose-dependent manner. MDA-MB-468 was arrested at the G2 phase at 7.5 uM. Induction of apoptosis was at 15 uM (MDA-MB-468) and 30uM (MDA-MB-231). In addition, protein expression for both cell lines, as determined by western blotting analysis, showed an increase of the cell cycle regulatory proteins cyclin B1 and CDK1 and the apoptotic protein caspase-3. Results for this small sample size demonstrate efficacy of treatment for both racial populations. Further studies analyzing the effect of Annonacin on additional diverse TNBC cell lines, organoids, and patient derived xenografts will advance the progression of alternative treatments for all patients with TNBC. Citation Format: Jennifer Hernandez-Torres, Kelly Argueta Guzman. Analyzing the effect of annonacin on diverse breast cancer cell lines as a potential therapeutic agent for triple negative breast cancer [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C026.

Discussion of spinal cord neurons apoptosis and neuroprotection mechanism of NGF gene transfection mediated by recombinant adenovirus in EAE mice

To investigate the spinal cord neuron apoptosis and neuroprotective mechanism of nerve growth factorganismsor (NGF) gene mediated by recombinant adenovirus (Ad-NGF) via peripheral transfection in mice with experimental autoimmune encephalomyelitis (EAE). Forty healthy female C57BL/6 mice were randomly divided into a control group, adenovirus (AdV) group, EAE group, and Ad-NGF transfection group; the control group received no treatment; the AdV group received adenovirus injection via the tail vein; the EAE and Ad-NGF transfection groups were induced with experimental autoimmune encephalomyelitis (EAE) using myelin oligodendrocyte glycoprotein 35–55 (MOG35-55), Ad-NGF transfection group received Ad-NGF injection via the tail vein, and daily neurological impairment scores were obtained. AQThe TUNEL method was employed to observe spinal neuron apoptosis in each group of mice; protein immunoblotting (western blot) and RT-PCR were used to measure NGF levels in the spinal cord tissues of each group, and western blotting was used to assess levels of cleaved caspase-3, Bax, and Bcl-2. ELISA and RT-PCR were employed to detect protein and mRNA levels of neuron-specific enolase (NSE) in spinal cord tissues, respectively. The control group and AdV mice did not develop symptoms. Compared to the EAE group, in the Ad-NGF transfection group, neurological function scores, TUNEL-positive cell counts, the ratio of NeuN + TUNEL to NeuN, levels of Bax and cleaved caspase-3 apoptotic proteins were significantly reduced, while Bcl-2 protein expression was increased. Expression levels of NGF, NGF-mRNA, NSE, and NSE-mRNA in spinal cord tissues were significantly elevated (P < 0.01). Immunofluorescence labeling revealed a significant punctate aggregation of apoptotic cells in spinal neurons of the EAE group, while the aggregation phenomenon was less pronounced in the Ad-NGF transfection group. Ad-NGF transfected by the periphery has a protective effect on spinal cord neurons in EAE mice by up-regulation NGF level, down-regulating apoptotic protein Caspase-3 in spinal cord neurons, inhibiting spinal cord neuron apoptosis and promoting NSE expression.

The effect of repetitive magnetic stimulation on neuronal apoptosis and PI3K/Akt protein expression in rats with incomplete spinal cord injury.

The pathological and physiological process of spinal cord injury is complex, and there is currently no effective treatment method. Magnetic stimulation is an emerging electromagnetic therapy method in recent years, and studies have shown its potential to reduce cell apoptosis. This study used an improved Allen's method to replicate an incomplete spinal cord injury rat model, and repetitive magnetic stimulation (rMS) intervention was performed on the rats for 21 days. The research plan consists of two parts. The first part aims to observe the effects of rMS on motor function and neuronal cell apoptosis in rats. The Basso-Beattie-Bresnahan (BBB) score results indicate that rMS promotes the recovery of motor function in rats; H&E staining showed that rMS improved spinal cord structural damage and inflammatory infiltration; TUNEL and NeuN staining suggest that rMS can reduce cell apoptosis and promote neuronal cell survival. The second part aims to explore the mechanism of action of rMS. Immunofluorescence staining showed that after rMS intervention, the positive counts of PI3K and Akt increased, whereas the positive counts of caspase-3 decreased. Western blot showed that after rMS intervention, the expression of phospho-phosphatidylinositol-3 kinase (p-PI3K)/PI3K, phospho (p)-Akt/Akt, and Bcl-2 increased, whereas the expression of Bcl-2-associated X protein(Bax) and caspase-3 decreased. In summary, rMS can significantly reduce cell apoptosis in the damaged spinal cord and promote neuronal cell survival. Its mechanism of action may be related to promoting the expression of PI3K/Akt pathway proteins, upregulating the antiapoptotic protein Bcl-2, downregulating the proapoptotic protein Bax, and thereby inhibiting the expression of apoptotic protein caspase-3. NEW & NOTEWORTHY Spinal cord injury is a serious disabling central nervous system disease. Recently, research on magnetic stimulation therapy for spinal cord injury has been increasing, and its potential has gradually attracted the attention of experts. This study found that repetitive magnetic stimulation (rMS) can improve motor function and reduce neuronal apoptosis in spinal cord injury rats. The mechanism may be related to increasing the expression of phosphatidylinositol-3 kinase (PI3K)/Akt protein, thereby inhibiting cell apoptosis and promoting neuronal survival.

Dopaminergic neurons lacking Caspase-3 avoid apoptosis but undergo necrosis after MPTP treatment inducing a Galectin-3-dependent selective microglial phagocytic response

Parkinson’s Disease (PD) is a progressive neurodegenerative disorder characterized by the loss of dopaminergic neurons in the Substantia nigra pars compacta (SNpc). Apoptosis is thought to play a critical role in the progression of PD, and thus understanding the effects of antiapoptotic strategies is crucial for developing potential therapies. In this study, we developed a unique genetic model to selectively delete Casp3, the gene encoding the apoptotic protein caspase-3, in dopaminergic neurons (TH-C3KO) and investigated its effects in response to a subacute regime of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration, which is known to trigger apoptotic loss of SNpc dopaminergic neurons. We found that Casp3 deletion did not protect the dopaminergic system in the long term. Instead, we observed a switch in the cell death pathway from apoptosis in wild-type mice to necrosis in TH-C3KO mice. Notably, we did not find any evidence of necroptosis in our model or in in vitro experiments using primary dopaminergic cultures exposed to 1-methyl-4-phenylpyridinium in the presence of pan-caspase/caspase-8 inhibitors. Furthermore, we detected an exacerbated microglial response in the ventral mesencephalon of TH-C3KO mice in response to MPTP, which mimicked the microglia neurodegenerative phenotype (MGnD). Under these conditions, it was evident the presence of numerous microglial phagocytic cups wrapping around apparently viable dopaminergic cell bodies that were inherently associated with galectin-3 expression. We provide evidence that microglia exhibit phagocytic activity towards both dead and stressed viable dopaminergic neurons through a galectin-3-dependent mechanism. Overall, our findings suggest that inhibiting apoptosis is not a beneficial strategy for treating PD. Instead, targeting galectin-3 and modulating microglial response may be more promising approaches for slowing PD progression.

Oral Administration of Carotenoid-Rich Dunaliella salina Powder Inhibits Colon Carcinogenesis via Modulation of Wnt/β-catenin Signaling Cascades in a Rat Model.

The present study aims to investigate the oral therapeutic and molecular role of carotenoid-rich Dunaliella salina powder (DSP) against 1,2-dimethylhydrazine (DMH)-triggered colon carcinogenesis. In this study, thirty six male Wistar rats were categorized into six distinct groups (G1-G6): G1 group with no intervention, G2 group received only DSP (1000mg/kg), G3 group received only DMH carcinogen (20mg/kg), and G4-G6 group received both DMH and DSP at various phases (pre-initiation, post-initiation and entire phases) for 32weeks. Body weight, tumor incidence, tumor volume, histopathological examination, antioxidants, and detoxification enzymes activities were analyzed in the experimental rats. In addition, the protein expression profile of components involved in the Wnt/β-catenin signaling pathway was determined by western blot analysis. Matrix metalloproteinases (MMP-7 and MMP-9), proliferation marker (PCNA), and pro-apoptotic (Bcl-2 and Bax) proteins were analyzed using immunohistochemistry. Colorimetric assay was used to determine the levels of anti-inflammatory (iNOS and COX-2) and apoptotic proteins (Caspase-3 and Caspase-9). Results showed that concomitant administration of DSP with DMH significantly reduced tumor progression and prevented colon carcinogenesis in rats. However, treatment with DSP before or after DMH exposure did not significantly prevent colon carcinogenesis. DMH and DSP treatment group showed increased activities of antioxidant enzymes with significant reduction in the oxidative stress. Additionally, the detoxification enzymes and colonic histopathology of those rats were restored to that of control rats. The administration of DSP to rats exposed to DMH exhibited antitumor effects via inhibition of the Wnt/β-catenin signaling pathway with induced apoptosis through the Bcl-2/Bax/caspases signaling cascades. Moreover, the same group also showed significant anti-inflammatory activity via regulating iNOS and COX-2 biomarkers. Our findings revealed molecular chemopreventive activity of carotenoid-rich DSP through regulating Wnt/beta-catenin and intrinsic apoptotic pathways. Thus, DSP is propound to function as a potent antioxidant, anti-proliferative, and anti-inflammatory therapeutic agent against colon carcinogenesis.

Synergistic effects of plasma-activated medium in combination with Baicalin against neuronal damage

Neurodegenerative disorders are chronic conditions that progressively damage and destroy parts of the nervous system, and are currently considered permanent and incurable. Alternative strategies capable of effectively healing neuronal damage have been actively pursued. Here, we report the neuroprotective effects of baicalin (BA) combined with plasma-activated medium (PAM) against glutamate-induced excitotoxicity in SH-SY5Y cells. Through in vitro assays, the cell viability, inflammation, apoptosis, and oxidative stress were evaluated. The co-application of BA and PAM significantly enhanced cell viability, reduced pro-inflammatory markers (TNF-α and NF-κB), decreased apoptotic proteins (Bax and Caspase-3) and boosted antioxidative defenses (increased SOD activity and lowered ROS levels). This study confirms the potential of combining BA with PAM as an effective therapeutic strategy for mitigating the effects of excitotoxicity. PAM is a promising adjunct and potential drug delivery method in neuroprotective therapy, providing a new avenue for developing treatments for diseases characterized by neuronal damage.

Solanine Inhibits Proliferation and Angiogenesis and Induces Apoptosis through Modulation of EGFR Signaling in KB-ChR-8-5 Multidrug-Resistant Oral Cancer Cells.

Background: The most important factors contributing to multi-drug resistance in oral cancer include overexpression of the EGFR protein and the downstream malignancy regulators that are associated with it. This study investigates the impact of solanine on inflammation, proliferation, and angiogenesis inhibition in multidrug-resistant oral cancer KB-Chr-8-5 cells through inhibition of the EGFR/PI3K/Akt/NF-κB signaling pathway. Methods: Cell viability was assessed using an MTT assay to evaluate cytotoxic effects. Production of reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨM), and AO/EtBr staining were analyzed to assess apoptosis and mitochondrial dysfunction. Western blotting was employed to examine protein expression related to angiogenesis, apoptosis, and signaling pathways. Experiments were conducted in triplicate. Results: Solanine treatment at concentrations of 10, 20, and 30 μM significantly increased ROS production, which is indicative of its antioxidant properties. This increase was associated with decreased mitochondrial membrane potential (ΔΨM) with p < 0.05, suggesting mitochondrial dysfunction. Inhibition of EGFR led to reduced activity of PI3K, Akt, and NF-κB, resulting in decreased expression of iNOS, IL-6, Cyclin D1, PCNA, VEGF, Mcl-1, and HIF-1α and increased levels of the apoptotic proteins Bax, caspase-9, and caspase-3. These changes collectively inhibited the growth of multidrug-resistant (MDR) cancer cells. Conclusions: Solanine acts as a potent disruptor of cellular processes by inhibiting the EGFR-mediated PI3K/Akt/NF-κB signaling pathway. These results suggest that solanine holds promise as a potential preventive or therapeutic agent against multidrug-resistant cancers.

Investigation of the Antarctic krill tripeptide Phe-Pro-Phe (FPF) for its antioxidant properties and mechanism

Oxidative stress causes damage to cells and brain injury-associated disorders. The study aimed to investigate how the tripeptide Phe-Pro-Phe (FPF), derived from an antioxidant-rich Antarctic krill peptide Ser-Ser-Asp-Ala-Phe-Phe-Pro-Phe-Arg (SSDAFFPFR), could protect against oxidative stress caused by scopolamine. Utilizing mouse and cell models to induce oxidative stress injury with scopolamine, coupled with Fourier transform infrared (FTIR) microscopy, immunofluorescence, and mitochondrial function assays, the study demonstrated that FPF significantly reduced malondialdehyde (MDA) and reactive oxygen species (ROS) levels, as well as decreasing the expression level of the apoptotic protein Caspase-3, while also increasing superoxide dismutase (SOD) activity and unsaturated lipid levels. Furthermore, FPF significantly improved mitochondrial aerobic respiration oxygen consumption rate (OCR) and anaerobic glycolytic metabolism extracellular acidification rate (ECAR) in mouse hippocampal neuron (HT22) cells. The results indicated that FPF protected against scopolamine-induced oxidative stress by regulating anti-oxidative activity, enhancing Caspase-3 expression, increasing unsaturated lipid levels, and improving cellular energy metabolism.

Synthesis, X-ray, Hirshfeld combined with DFT, anticancer effects with molecular docking confirmation of (E)-4-cyanobenzaldehyde oxime and (Z)–N-methyl-C-4-substituted phenyl aldonitrones

(E)-4-cyanobenzaldehyde oxime 1 and (Z)–N-methyl-C-4-substituted phenyl aldonitrones 2 and 3 were synthesised and fully characterised. With the help of Hirshfeld calculations, the molecular packing of 1 is found to be controlled by short N…H contacts (28.3%) in addition to the aromatic π-π stacking where the%C…C interactions is calculated to be 9.7%. In this case, the second most abundant interaction is H…H contact (28.1%). For 2, the N…H (24.4%) and O…H (13.8%) contacts are the most important while the most abundant interaction is H…H contacts (36.0%). DFT calculations were utilised to explore the structural and electronic parameters of both compounds at their optimised geometries which were found in good agreement with the X-ray structures. The structure of 2 (6.9829 Debye) is more polar than 1 (4.5282 Debye). The compounds 1–3 reduced the growth of MCF-7 and T47D. Compound 1 which has the lowest IC50 values against the MCF-7 and T47D cell lines, is thought to be the most promising applicant as an anticancer drug. The in-silico combination of compounds 1–3 with proliferative proteins PIK3 and CDK5, autophagic proteins mTOR and beclin, antiapoptotic proteins BCL2, and apoptotic proteins caspase 3 was examined. Compound 3 binds to PIK3, CDK5, mTOR, Beclin, BCL2 and caspase 3, and displays higher free energy bindings of −5.12, −5.45, −4.923, −5.72, −4.45 and −4.73 kcal/mol, which suggested that these proteins had the strongest inhibitory or enhanced effects. The findings show that compound 3 was the most well-docked chemical with the greatest IC50. Compound 3 impacted the autophagy, apoptotic and mitotic proteins of cell growth.

CD38 Deficiency Protects Mouse Retinal Ganglion Cells Through Activating the NAD+/Sirt1 Pathway in Ischemia-Reperfusion and Optic Nerve Crush Models.

The purpose of this study was to investigate the protective effect of CD38 deletion on retinal ganglion cells (RGCs) in a mouse retinal ischemia/reperfusion (I/R) model and an optic nerve crush (ONC) model, and to elucidate the underlying molecular mechanisms. Retinal I/R and ONC models were constructed in mice. PCR was used to identify the deletion of CD38 gene in mice, hematoxylin and eosin (H&E) staining was used to evaluate the changes in retinal morphology, and electroretinogram (ERG) was used to evaluate the changes in retinal function. The survival of RGCs and activation of retinal macroglia were evaluated by immunofluorescence staining. The expression of Sirt1, CD38, Ac-p65, Ac-p53, TNF-α, IL-1β, and Caspase3 proteins in the retina was further evaluated by protein imprinting. In retinal I/R and ONC models, CD38 deficiency reduced the loss of RGCs and activation of macroglia and protected the retinal function. CD38 deficiency increased the concentration of NAD+, reduced the degree of acetylation of NF-κB p65 and p53, and reduced expression of the downstream inflammatory cytokines TNFα, IL-1β, and apoptotic protein Caspase3 in the retina in the ONC model. Intraperitoneal injection of the Sirt1 inhibitor EX-527 partially counteracted the effects of CD38 deficiency, suggesting that CD38 deficiency acts at least in part through the NAD+/Sirt1 pathway. CD38 plays an important role in the pathogenesis of retinal I/R and ONC injury. CD38 deletion protects RGCs by attenuating inflammatory responses and apoptosis through the NAD+/Sirt1 pathway.

Curcumin synergistically augments the chemotherapeutic activity of Doxorubicin in prostate cancer cells

Objective Prostate cancer is one of the most commonly diagnosed cancer types in men and many people die every year due to recurring or acquiring aggressive forms of prostate cancer. Numerous chemotherapeutics such as paclitaxel and doxorubicin are commonly used in the treatment of prostate cancer. However, acquired resistance to chemotherapeutics and broad systemic side effects substantially limit their efficacy. Curcumin is one of the most examined phytochemicals of the herbal remedy turmeric. Herein, we aimed to investigate the synergistic capability of curcumin on doxorubicin in prostate cancer cells. Method The human adenocarcinoma cell line LNCaP was used in cell culture studies. Cell viability was examined by WST-1 assay. The protein expression levels of Beclin1, p62/SQSTM1, LC3-I/II, Hrd1, gp78, polyubiquitin, PERK, eIF2, phospho-(Ser51) eIF2, IRE1, XBP-1s, PARP-1, caspase-3, AR, PSA, c-Myc, E-cadherin, N-cadherin and VEGF-A were investigated by immunoblotting assay. Results Our data indicated that co-administration of curcumin with doxorubicin significantly improved the cytotoxic effect of doxorubicin in LNCaP cells. Also, the combination of curcumin and doxorubicin reduced the autophagic flux and remarkably induced endoplasmic reticulum-associated-degradation (ERAD) and unfolded protein response (UPR) signaling. Also, activation of apoptotic proteins PARP-1 and caspase-3 were strongly enhanced by combined treatment in a dose-dependent manner. Moreover, combined treatment markedly decreased AR, PSA, c-Myc and VEGF-A levels. Additionally, the epithelial-mesenchymal transition (EMT) was reduced by decreasing N-cadherin and increasing E-cadherin protein levels. Conclusion Present data strongly suggest that curcumin synergistically improves the anti-cancer features of doxorubicin in prostate cancer cells.

Assessment of the in Vitro Effects of Folate Core-Shell Conjugated Iron Oxide Nanoparticles as a Potential Agent for Acute Leukemia Treatment.

There is a growing need to comprehend the potential outcomes of nanoparticles (NPs) on human well-being, including their potential for detecting and treating leukemia. This study examined the role of iron folate core-shell and iron oxide nanoparticles in inducing apoptosis and altering the expression of the B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X-protein (Bax), and Caspase-3 genes in leukemia cells. The obtained iron oxide and iron folate core-shell nanoparticles were analyzed using a variety of analytical techniques, including ultraviolet-visible (UV-Vis) absorption spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), dynamic light scattering (DLS), zeta potential, and transmission electron microscopy (TEM). Additionally, FTIR and UV-Vis were used to characterize doxorubicin. The MTT test was utilized to investigate the cytotoxicity of iron oxide and iron folate core-shell nanoparticles. The expression of the apoptotic signaling proteins Bcl-2, Bax, and Caspase-3 was evaluated using the real-time reverse transcription polymerase chain reaction (RT-qPCR) method. Additionally, flow cytometry was performed to gauge the degrees of necrosis and apoptosis. UV-Visible spectroscopy analysis showed that the generated iron oxide and iron folate core-shell NPs had a distinctive absorption curve in the 250-300 nm wavelength range. The XRD peaks were also discovered to index the spherical form with a size of less than 50 nm, which validated the crystal structure. The FTIR analysis determined the bonds and functional groups at wavenumbers between 400 and 4000 cm-1. A viable leukemia treatment approach is a nanocomposite consisting of iron and an iron folate core-shell necessary for inhibiting and activating cancer cell death. The nearly resistant apoptosis in the CCRF-CEM cells may have resulted from upregulating Bax and Casepase-3 while downregulating Bcl-2 expression. Our study documents the successful synthetization and characterization of iron oxide, which has excellent anticancer activities. A metal oxide conjugation with the nanoparticles' core-shell enhanced the effect against acute leukemia.

Protective Effect of Prunella vulgaris L. Whole Plant or Spicas Water Extracts on Hyperthyroidism with Liver-yang Hyperactivity Syndrome

Background: Prunella vulgaris L. (Lamiaceae) is a perennial medicinal plant commonly used to treat thyroid dysfunction, infectious hepatitis, and mastitis in north-east Asia and Europe. In China, there has been a historical change in the medicinal use of P. vulgaris from the use of whole plants to the use of the spicas. Purpose: This study aimed to evaluate the protective effects of whole plant P. vulgaris water extracts and spicas on hyperthyroidism with liver-yang hyperactivity syndrome and to preliminarily explain the mechanism of action of these water extracts. Methods: Rats were induced with aconite decoction and levothyroxine and treated with P. vulgaris whole plant and spicas water extract for 28 days at the same time. Body weight, food intake, and rectal temperature were monitored. The serum thyroid function indicators thyroid stimulating hormone (TSH), thyroid hormone-free triiodothyronine (FT3) and free thyroxine (FT4), liver function index alanine aminotransferase (ALT) and aspartate aminotransferase (AST), oxidative damage index malondialdehyde (MDA), and superoxide dismutase (SOD) were detected. Liver and thyroid tissues were taken for sectioning and observation. The liver inflammatory factors human tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by immunohistochemistry, and the expression levels of the apoptotic proteins caspase-3 and B-cell lymphoma-2 (Bcl-2) were detected. Results: Both P. vulgaris whole plant and spica water extract groups exhibited markedly reduced FT3 and FT4 contents, increased the TSH content, and decreased ALT and AST activities. Furthermore, these agents improve thyroid and liver lesion tissue; reduce MDA content; increase SOD activity; reduce IL-6, TNF-α, and caspase-3 expression; and increase Bcl-2 expression. Conclusion: The whole plant and spicas water extracts of P. vulgaris have obvious protective effects on hyperthyroidism and hyperactivity in liver yang, and the protective effect of the water extract of spicas is greater than that of the whole plant water extract.

TonEBP/NFAT5 expression is associated with cisplatin resistance and migration in macrophage-induced A549 cells

BackgroundMacrophages promote angiogenesis, metastasis, and drug resistance in several cancers. Similarly, TonEBP/NFAT5 induces metastasis in renal carcinoma and colon cancer cells. However, the role of this transcription factor and that of macrophages in lung cancer cells remains unclear. Therefore, this study investigated the effects of macrophages and TonEBP/NFAT5 expression on cisplatin resistance and migration in A549 lung adenocarcinoma cells.ResultsA549 cells were cultured alone or indirectly co-cultured with THP-1-derived macrophages using a transwell culture chamber. Cisplatin-induced cell death was markedly decreased and migration increased in co-cultured A549 cells. Macrophage-conditioned media (CM) showed a similar effect on drug resistance and migration. Cisplatin-induced apoptosis, DNA fragmentation, and cleaved apoptotic proteins PARP and caspase-3 were markedly reduced in macrophage CM-induced A549 cells. Here, ERK, p38, JNK, and NF-κB activities were increased by macrophage CM. Furthermore, the proteins involved in cisplatin resistance and cancer cell migration were identified using specific inhibitors of each protein. ERK and NF-κB inhibition considerably reduced cisplatin resistance. The increase in macrophage CM-induced migration was partially reduced by treatment with ERK, JNK, and NF-κB inhibitors. TonEBP/NFAT5 expression was increased by macrophages, resulting in increased cisplatin resistance, cell migration, and invasion. Moreover, RNAi-mediated knockdown of TonEBP/NFAT5 reduced cisplatin resistance, migration, and invasion in macrophage CM-induced A549 cells.ConclusionsThese findings demonstrate that paracrine factors secreted from macrophages can change A549 cells, resulting in the induction of drug resistance against cisplatin and migration. In addition, the TonEBP/NFAT5 ratio, increased by macrophages, is an important regulator of the malignant transformation of cells.

PEI, a new transfection method, augments the inhibitory effect of RBM5 on prostate cancer

PEI is a cationic polymer, serving as a non-viral transfection carrier grounded in nanotechnology that enhances transfection efficiency via the proton sponge effect. RBM5 is an RNA-binding protein that can inhibit tumor development. This study involved the transfection of RBM5 in prostate cancer cells with PEI, Lipo2000, and their combination. Transwell and wound healing assays were used to observe invasion and migration of prostate cancer cells and flow cytometry was used to observe the apoptosis. Detect the expression of invasion and migration-related protein MMP9 through western blotting experiment. An activity detection kit was used to detect the activity of apoptotic protein caspase-3. We found that there was no significant difference in transfection efficiency when PEI and Lipo2000 were used alone but it significantly improved when they are combined. RBM5 reduced invasion, migration, and proliferation of prostate cancer and enhanced apoptosis. MMP9 expression was reduced, and the activity of caspase-3 was increased. PEI transfection could improve the inhibition of RBM5 on tumors more than Lipo2000. The inhibitory effect is more obvious when the two are used together. RBM5 transfected with PEI can amplify its inhibitory effect on prostate cancer, and this effect is more evident when combined with Lipo2000.

Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway.

Gynecologic tract melanoma is a malignant tumor with poor prognosis. Because of the low survival rate and the lack of a standard treatment protocol related to this condition, the investigation of the mechanisms underlying melanoma progression is crucial to achieve advancements in the relevant gynecological surgery and treatment. Mitochondrial transfer between adjacent cells in the tumor microenvironment regulates tumor progression. This study investigated the effects of endothelial mitochondria on the growth of melanoma cells and the activation of specific signal transduction pathways following mitochondrial transplantation. Mitochondria were isolated from endothelial cells (ECs) and transplanted into B16F10 melanoma cells, resulting in the upregulation of proteins associated with tumor growth. Furthermore, enhanced antioxidation and mitochondrial homeostasis mediated by the Sirt1-PGC-1α-Nrf2-HO-1 pathway were observed, along with the inhibition of apoptotic protein caspase-3. Finally, the transplantation of endothelial mitochondria into B16F10 cells promoted tumor growth and increased M2-type macrophages through Nrf2/HO-1-mediated pathways in a xenograft animal model. In summary, the introduction of exogenous mitochondria from ECs into melanoma cells promoted tumor growth, indicating the role of mitochondrial transfer by stromal cells in modulating a tumor's phenotype. These results provide valuable insights into the role of mitochondrial transfer and provide potential targets for gynecological melanoma treatment.

A peptide encoded by the circular form of the SHPRH gene induces apoptosis in neuroblastoma cells.

Circular RNAs (circRNAs) and their derived peptides represent largely unchartered areas in cellular biology, with many potential roles yet to be discovered. This study aimed to elucidate the role and molecular interactions of circSHPRH and its peptide derivative SHPRH-146aa in the pathogenesis of neuroblastoma (NB). NB samples in the GSE102285 dataset were analyzed to measure circSHPRH expression, followed by in vitro experiments for validation. The role of SHPRH-146aa in NB cell proliferation, migration, and invasion was then examined, and luciferase activity assay was performed after SHPRH-146aa and RUNX1 transfection. Finally, the regulation of NB cell apoptosis by SHPRH-146aa combined with NFKBIA was tested. The GSE102285 dataset indicated overexpression of circSHPRH in NB samples, further supported by in vitro findings. Overexpression of circ-SHPRH and SHPRH-146aa inhibited proliferation, migration, and invasion of NB cells. A significant increase in apoptosis was observed, with upregulation of Caspase-3 and downregulation of Bcl-2. Furthermore, the peptide derivative SHPRH-146aa, derived from circSHPRH, suppressed NB cell malignancy traits, suggesting its role as a therapeutic target. A direct interaction between SHPRH-146aa and the transcription factor RUNX1 was identified, subsequently leading to increased NFKBIA expression. Notably, NFKBIA knockdown inhibited the pro-apoptotic effect of SHPRH-146aa on NB cells. The study demonstrates that circ-SHPRH and SHPRH-146aa play significant roles in inhibiting the malignant progression of NB. They induce apoptosis primarily by modulating key apoptotic proteins Caspase-3 and Bcl-2, a process that appears to be regulated by NFKBIA. The SHPRH-146aa-RUNX1 interaction further elucidates a novel pathway in the regulation of apoptosis in NB. These findings indicate that circ-SHPRH and its derived peptide SHPRH-146aa could be potential therapeutic targets for NB treatment.