Design, synthesis and evaluation of Nur77 modulators for lysosome dysfunction-driven therapy of gastric cancer.

The role of mTORC2 in the in vitro neurotoxicity of Parkinsonian mimetics 6-OHDA and MPP.

Breast cancer remains the most prevalent malignancy worldwide, with triple-negative breast cancer (TNBC) being particularly aggressive and resistant to standard therapies. Plant-derived compounds have emerged as promising alternatives due to selective cytotoxicity and reduced adverse effects. Portulaca grandiflora (Portulacaceae), ornamental succulent rich in flavonoids, phenolics, carotenoids, and sterols-phytochemicals commonly associated with anticancer activity-was evaluated for its pro-apoptotic and anti-metastatic potential. Three crude extracts, methanolic (PGM), ethanolic (PGE), and n-hexane (PGH), were initially screened for cytotoxicity, after which the two most active extracts, PGM and PGH, were selected for detailed mechanistic analysis. MTT assay was used for antiproliferative screening against MDA-MB-231 TNBC cells and non-tumorigenic MCF-10A breast epithelial cells. Apoptosis induction was assessed using sub-G1 cell cycle arrest, Annexin V/PI staining, DNA fragmentation, and caspase-3/7 activation assays. NF-κB signaling and associated survival proteins were evaluated through Western blot, luciferase reporter, and DNA-binding assays. Anti-migratory and anti-invasive effects were analyzed using wound healing and transwell invasion assays. PGM showed the lowest IC₅₀ (79.5µg/mL at 72h) with minimal toxicity to MCF-10A cells. Both PGM and PGH triggered sub-G1 accumulation and dose-dependent apoptosis, with PGM eliciting stronger caspase-3/7 activation, DNA fragmentation, and PARP cleavage. Notably, PGM inhibited NF-κB DNA-binding activity by ~ 60% compared to ~ 40% with PGH, reduced IκBα, TAK1, and Akt phosphorylation, and prevented nuclear p65 translocation, accompanied by downregulation of Bcl-2, Bcl-xL, XIAP, survivin, and Cyclin D1. Migration and invasion were significantly suppressed (p < 0.01), confirming anti-metastatic efficacy. Collectively, these findings demonstrate that P. grandiflora extracts-especially PGM-selectively induce apoptosis and impede metastasis in TNBC cells through suppression of NF-κB signaling activity (association-based evidence), supporting its potential as a phytochemical source for targeted breast cancer therapy. Because pathway-causality was not tested using NF-κB rescue or pharmacological inhibition controls, the mechanistic conclusions are presented as supportive associations rather than definitive proof. Future studies should focus on bioassay-guided compound purification, synergistic combination assessment, and in vivo validation for translational advancement.

Lung cancer remains the leading cause of both cancer incidence and mortality worldwide, rendering it a primary focus in oncology research. This pressing global health challenge has created an urgent demand for novel therapeutic agents with high efficacy and low toxicity. Owing to their distinctive chemical structures and wide range of biological activities, natural products have become essential resources in the development of medicines. Alepterolic acid is a natural diterpenoid from the plant Aleuritopteris argentea (Gmél.) Fée, a fern used in folk medicine in China. In this study, a total of 31 derivatives were gained by the incorporation of urea and piperazine groups into alepterolic acid. Among them, the inhibition rate of 21 compounds against A549 cells exceeded 50% at 10 μM. Specifically, the IC50 values of compounds 15o and 15u against A549 cells were 1.8 ± 0.3 and 1.5 ± 0.2 μM, respectively. Moreover, it was found that the proliferation and migration of A549 cells were inhibited remarkably by both compounds 15o and 15u as the cell cycle was arrested at the G1 phase. At the molecular level, compounds 15o and 15u exhibited the ability to induce cleavage and activation of caspase-3 and caspase-9, promote PARP-1 cleavage, downregulate Bcl-2 expression, and upregulate Bax protein levels. The docking simulation results suggested that compounds 15o and 15u could stably bind to caspase-3, Bax, Bcl-2, caspase-9, and PARP-1. The physicochemical property/ADMET evaluation demonstrated that the physicochemical characteristics of compound 15o were indicative of an enhanced bioavailability. To sum up, urea- and piperazine-functionalized alepterolic acid derivatives possess significant anti-cancer activities and are worthy of further research.

Pristimerin drives ROS-dependent apoptosis in cutaneous T-cell lymphoma via inhibition of the AKT-SKP2 axis.

Cutaneous T-cell lymphoma (CTCL) is a rare non-Hodgkin lymphoma with limited durable treatment options and poor overall survival, underscoring the need for new therapeutic approaches. Sanguinarine (SNG), a natural benzophenanthridine alkaloid, has demonstrated anticancer activity in several malignancies, but its potential role in CTCL remains unexplored. We hypothesized that SNG induces oxidative stress and mitochondrial dysfunction, leading to the inhibition of the PI3K/AKT/GSK3 pathway and apoptosis in CTCL cells. Invitro experiments were performed using HH and H9 CTCL cell lines. Cytotoxicity, apoptosis, and mitochondrial function were evaluated by cell viability assays, caspase activation, PARP cleavage, Bax/Bcl-2 ratio, mitochondrial membrane depolarization, and cytochrome c release. ROS generation and glutathione depletion were measured with and without N-acetyl cysteine (NAC) rescue. Western blotting assessed modulation of the AKT/GSK3α/β/mTOR pathway and expression of anti-apoptotic proteins (XIAP, cIAPs, Mcl-1). Synergistic effects with bortezomib (BTZ) were analyzed. Network pharmacology, molecular docking, molecular dynamics (MD) simulations, and binding free energy (BFE) calculations were used to identify SNG's molecular targets. SNG reduced CTCL cell viability (IC₅₀ < 5 μM) and triggered mitochondrial-mediated apoptosis, accompanied by Bax/Bcl-2 modulation, ΔΨm loss, cytochrome c release, and caspase-9/3 activation. ROS accumulation and glutathione depletion contributed to cytotoxicity, effects that were reversed by NAC. SNG suppressed AKT/GSK3/mTOR signaling and downregulated anti-apoptotic proteins. Notably, SNG enhanced the anticancer activity of BTZ in combination studies. Computational analyses supported AKT and Bcl-2 as key binding targets. SNG exerts potent anticancer effects in CTCL by inducing ROS-dependent mitochondrial apoptosis and inhibiting the PI3K/AKT/GSK3 signaling pathway. Its synergy with BTZ and computational validation of AKT/Bcl-2 targeting underscore its potential as a novel therapeutic candidate for CTCL, warranting further preclinical investigation.

Discovery of novel sophocarpine derivatives as potential dual Bcl-2 and Mcl-1 inhibitors: design, synthesis and anti-hepatocellular carcinoma evaluation.

Endometriosis is a chronic, hormone-dependent disorder characterized by ectopic implantation of endometrial tissue, often accompanied by pain and infertility. Although the progesterone receptor modulator RU486 is effective for pain relief, its impact on lesion regression is limited, possibly due to apoptosis resistance and overexpression of PARP1 in endometriotic cells. This study aimed to evaluate the synergistic therapeutic efficacy of combining RU486 with the PARP inhibitor olaparib, focusing on reactivating p53-dependent apoptosis in endometriotic lesions. Primary ectopic endometrial stromal cells (EESCs) were used to assess the effects of RU486 and olaparib on cytotoxicity, apoptosis, DNA damage, mitochondrial membrane potential, and cell migration. Mechanistic investigations included flow cytometry, western blotting, RT-qPCR, immunofluorescence, and comet assays. A murine endometriosis model was established to evaluate in vivo lesion suppression, pain behaviour, hormone levels, and treatment safety following combination therapy. RU486 and olaparib exhibited strong synergy at a 1:4 ratio, significantly enhancing apoptosis in EESCs by coactivating p53 through distinct mechanisms-transcriptional up-regulation by RU486 and posttranslational stabilization by olaparib. This synergy intensified Bax/Bcl-2-mediated mitochondrial dysfunction, caspase-3 activation, and PARP1 cleavage. In vivo, the combination therapy achieved a 77.5% lesion weight regression, significantly exceeding monotherapies, while maintaining robust analgesic effects. Histological and molecular analyses confirmed DNA damage and enhanced apoptosis in lesions. The combination medication enhanced apoptotic sensitivity in endometriosis by restoring p53 activity through targeting hormonal signalling and DNA repair pathways. This strategy offered strong potential for clinical translation.

Febuxostat, a selective xanthine oxidase inhibitor, was modified via 1,2,3-triazole-based molecular hybridization to generate a series of derivatives with potential anticancer activity. Among them, compounds 8c and 8e exhibited significant antitumor effects across multiple cancer cell lines, particularly in HepG2 cells, with IC50 values of 6.96 ± 1.76 and 8.56 ± 1.93 μM, respectively. Colony formation assays confirmed that both compounds markedly inhibited tumor cell proliferation. Annexin V-FITC/PI and TUNEL assays further demonstrated that 8c and 8e induced DNA damage and apoptosis in HepG2 cells, as shown by increased γ-H2AX expression and enhanced DNA strand breaks. They also activated intrinsic apoptotic pathways, including the cleavage and activation of caspase-9, caspase-3, and PARP. In vivo, nude mouse xenograft experiments validated their antitumor efficacy, with elevated cleaved caspase-3 levels in tumor tissues. Collectively, these findings indicate that 8c and 8e possess strong therapeutic potential against liver cancer and exhibit promising efficacy and safety profiles.

Design, synthesis and biological evaluation of novel chalcone analogs against castration-resistant prostate cancer.

p53-R248W evades doxorubicin mediated cell death through collective alteration in TOP2A and CYP1A1 genes in cancer cells.

This study aimed to determine whether repetitive transcranial magnetic stimulation (rTMS) is associated with changes in motor recovery and apoptosis-related pathways after spinal cord injury (SCI), and to characterize the temporal dependence of these effects. A rat model of SCI was established using Allen's method, and rTMS was initiated on day 2 post-injury. Western blot and immunofluorescence were used to quantify apoptosis-related execution-phase proteins, including caspase-3, cleaved caspase-3, caspase-7, PARP1, and cleaved PARP1, and to assess their cellular distribution. Double immunofluorescence of NeuN and cleaved caspase-3 was performed to identify the localization of apoptotic signals. TUNEL staining, HE staining, and BBB scores were used to assess apoptotic cell death, tissue injury, and motor recovery. rTMS was associated with time-dependent changes, with limited effects at early stages but reduced apoptosis-related signaling and improved tissue preservation and functional recovery at later time points. Co-localization of NeuN and cleaved caspase-3 indicated that apoptotic signaling was localized in neurons. These findings suggest that rTMS is associated with reduced activation of apoptosis-related execution-phase signaling after SCI in a time-dependent manner, together with enhanced tissue preservation and functional recovery. The caspase-3/caspase-7/PARP1 cleavage axis may be involved in the biological changes associated with rTMS treatment after SCI.

Esophageal squamous cell carcinoma (ESCC) is characterized by intrinsic radioresistance, primarily mediated by hyperactivation of DNA damage response (DDR) pathways. This repair capacity significantly diminishes radiotherapy efficacy, contributing to poor patient outcomes. Targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a master regulator of non-homologous end joining (NHEJ), represents a promising strategy to overcome radioresistance. Immunohistochemistry (IHC) was performed on paired biopsy specimens from esophageal squamous cell carcinoma (ESCC) patients (n = 20) collected before neoadjuvant radiotherapy and after surgical resection. Invitro, ESCC cell lines (TE13 and Eca9706) were co-treated with DNA-dependent protein kinase catalytic subunit (DNA-PKcs) inhibitors (AZD7648 or NU7741) or PRKDC-targeting siRNA plus irradiation. Subsequent functional assays were conducted to assess cell viability, apoptosis, cell cycle distribution, and underlying molecular mechanisms. Invivo, the combined efficacy of AZD7648 and irradiation was evaluated in xenograft tumor models. Phosphorylated DNA-PKcs (Ser2056) in ESCC tissues was significantly elevated following irradiation. Inhibition of DNA-PKcs markedly enhanced radiosensitivity in both ESCC cell lines and xenograft models, correlating with prolonged G2/M arrest and increased apoptosis. Mechanistically, the combined therapy activated the ATM (Ser1981)-Chk2 (Thr68) axis and concurrently enhanced phosphorylation of Chk1 at Ser345, reinforcing G2/M arrest through Wee1 kinase-mediated inhibitory phosphorylation of CDC2 at Tyr15. Consequently, irreparable DNA double-strand breaks (DSBs) accumulated, triggering Caspase-3-dependent cleavage of PARP and execution of the intrinsic apoptosis pathway. Our study reveals a novel mechanism by which DNA-PKcs inhibition augments radiosensitivity in ESCC. Targeting DNA-PKcs could represent a promising strategy to improve the efficacy of radiotherapy in ESCC, offering a potential therapeutic approach to overcome radioresistance.

Lactucin, a sesquiterpene lactone isolated from chicory (Cichorium intybus L.), demonstrates broad-spectrum anticancer activity across multiple malignancies, yet its mechanistic role in gastric cancer (GC) remains unexplored. This study aimed to elucidate the potential mechanism by which lactucin modulates GC cell proliferation and apoptosis. NUGC-3 and MKN-1 cells were used for in vitro studies, and xenograft tumor models were established using BALB/c nude mice for animal studies. To assess cell proliferation, cell cycle distribution, and apoptosis, CCK-8 assay and flow cytometry analyses were conducted, while cell morphology was examined under an inverted microscope. Levels of proteins associated with cell cycle, apoptosis, and mitogen-activated protein kinase (MAPK) signaling were determined by Western blotting. In vitro, lactucin significantly attenuated GC cell proliferation and induced alterations in their morphology. Lactucin induced cell cycle arrest in the G0/G1 phase by reducing protein levels of cyclins (cyclin B1 and cyclin D1) and cyclin-dependent kinases (CDKs), including CDK2 and CDK4. Moreover, lactucin promoted GC cell apoptosis in a dose-dependent manner by downregulating Bcl-2, upregulating Bax, and increasing cleavage of caspase-3 and PARP. In vivo, lactucin inhibited tumor growth and decreased Ki67 expression levels in xenograft tumors. Lactucin inhibited proliferation, induced cell cycle arrest and apoptosis in GC cells by inactivating the MAPK/p38 pathway.

Systemic toxicity and limited tumor selectivity remain major obstacles in HER2-positive breast cancer therapy with conventional chemotherapeutics. Extracellular vesicles (EVs) provide a promising delivery platform due to their intrinsic biocompatibility and intercellular communication. Here, we engineered EVs displaying the HER2-binding peptide P51 to create a peptide-mediated targeted delivery system for doxorubicin (Dox), aiming to enhance tumor specificity and apoptotic efficacy. EVs expressing the P51 peptide were generated by transient transfection of HEK-293 cells, followed by Dox loading to produce P51-EVDox. Cellular uptake and antitumor efficacy were evaluated in vitro and in vivo using HER2-overexpressing BT-474 breast cancer cells. RNA sequencing and apoptosis-related protein analyses, including cleaved caspase-3, PARP cleavage, and p53 expression, were performed to elucidate EV-mediated molecular mechanisms. P51-EVDox exhibited significantly enhanced binding and intracellular accumulation in HER2-positive cells compared with non-targeted EV-Dox. Transcriptomic analysis comparing free Dox and P51-EVDox revealed marked upregulation of thioredoxin-interacting protein (TXNIP). Consistently, protein-level analyses demonstrated increased activation of apoptosis-associated markers, including cleaved caspase-3, PARP cleavage, and p53, following P51-mediated EV delivery. These findings suggest activation of a ROS/TXNIP-driven apoptotic cascade. This study demonstrates that peptide-functionalized EVs represent a promising tumor-targeted chemotherapeutic platform for HER2-positive breast cancer. By integrating ligand-mediated cellular recognition with EV-based doxorubicin delivery, P51-EVDox activates the ROS/TXNIP signaling pathway, leading to enhanced tumor suppression and apoptosis.

Malignant mesothelioma is an aggressive cancer with limited therapeutic options, highlighting the need for novel strategies targeting metabolic vulnerabilities. Natural polyphenols have gained attention due to their ability to modulate cellular metabolism and apoptosis-related signaling pathways. In this study, we investigated the combined anticancer effects of quercetin (QUE) and arctigenin (ATG) in human malignant mesothelioma cells. QUE and ATG reduced the viability of MSTO-211H cells in a time-dependent manner, while non-malignant mesothelial MeT-5A cells showed relatively limited sensitivity under the tested conditions. Compared with single treatment, the combination treatment further enhanced growth inhibition, with combination index analysis suggesting a potential synergistic interaction. Co-treatment significantly decreased intracellular ATP levels and increased caspase-3/7 activity, suggesting metabolic stress-associated apoptotic responses. Annexin V analysis confirmed increased apoptotic cell populations following combination treatment. Western blot analysis demonstrated reduced expression of anti-apoptotic proteins Mcl-1 and Bcl-xL, along with increased cleavage of caspase-3 and PARP, consistent with involvement of intrinsic apoptosis-associated signaling pathways. In addition, increased phosphorylation of AMPK and altered expression of mitochondrial oxidative phosphorylation (OXPHOS) complex proteins were associated with potential alterations in mitochondrial respiratory protein expression. Collectively, these findings suggest that QUE and ATG co-treatment is associated with increased apoptotic cell death in malignant mesothelioma cells in association with metabolic stress–related mitochondrial functional alterations.

Inhibition of mTOR signaling pathway gains a lot of interests as promising strategies for cancer treatment. Recently, we have reported the anticancer potential of the other renieramycin T (RT) right-half compounds. The subsequent modifications were conducted to identify a novel compound capable of exerting anticancer activity through the inhibition of survival proteins and anti-apoptotic proteins. The right-half of RT analog, DH_27 was synthesized, and its activity was explored in this study. The cytotoxicity effects were defined with MTT assay and colony formation assay. The induction of apoptosis was elucidated through Hoechst33342/propidium iodide (PI) staining and Annexin V-FITC/PI staining, coupled with flow cytometry. Mitochondrial membrane potential was assessed using JC-1 staining to investigate mitochondrial-mediated apoptosis. An examination of apoptotic-related proteins and mTOR signaling proteins were evaluated. Binding affinity of DH_27 and mTOR and the stability of DH_27-stabilized protein-protein interactions were investigated using molecular docking and molecular dynamics simulation. DH_27 exhibited cytotoxic effects on NSCLC cells with an IC50 below 10 µM and hindered their ability to form colonies. Treatment with DH_27 induced apoptosis, as evidenced by reduced levels of the anti-apoptotic protein Bcl‑2 and increased cleavage of PARP and caspase-9. JC-1 staining revealed mitochondrial depolarization in DH_27-treated cells, further supporting apoptosis via the intrinsic pathway. Notably, DH_27 significantly downregulated p-mTOR/mTOR expression through stable interactions at both the allosteric and catalytic sites, leading to the suppression of downstream signaling pathways, including Akt and p85S6K. This study identifies DH_27 as a potent anticancer agent that inhibits mTOR through allosteric and catalytic mechanisms, suppressing Akt and p85S6K signaling. This leads to reduced cell growth and apoptosis, supporting DH_27 as a promising candidate for targeted anticancer therapy.

Background and aim Smp24 and Smp43 are cationic antimicrobial peptides from the Egyptian scorpion Scorpio maurus palmatus. Prior studies showed their cytotoxic effects on various cancer cell lines, including blood, liver, lung, and breast cancers. This study explored the mechanisms of Smp24 and Smp43 in glioma (U87MG) and prostate cancer (PC3) cells. Methods Cytotoxicity was assessed after 24-h treatment (2.5–40 µM) using CellTiter-Glo® for ATP levels and LDH assays for membrane integrity. At IC50 concentration, nuclear morphology was examined by Hoechst 33342/PI dual staining, and apoptosis by Annexin V/PI and cell cycle were analyzed by flow cytometry. Gene expression of apoptosis markers (CASP3, CASP8, CASP9, PARP1) was measured by RT-PCR. Protein activation of caspase-3 and PARP-1 was analyzed by Western blot, with caspase-3 involvement confirmed using the inhibitor Ac-DEVD-CHO. Key findings and conclusion Both peptides reduced viability and induced cytotoxicity in a dose-dependent manner, as evidenced by decreased ATP levels and increased LDH release. Morphological apoptosis features were observed with Hoechst/PI staining and confirmed by Annexin V/PI staining. In addition, PC3 cells only showed G2/M arrest. RT-PCR indicated upregulation of apoptosis markers, and Western blot confirmed caspase-3 activation and PARP-1 cleavage. Furthermore, Ac-DEVD-CHO attenuated peptide-induced cytotoxicity, supporting caspase-dependent apoptosis. Taken together, this study shows that Smp24 and Smp43 exhibit potent cytotoxicity against glioma and prostate cancer cells through caspase-dependent apoptosis, emphasizing their potential as the novel candidates for cancer therapy.

Natural product inspired scaffold hopping strategy: Identification of potent anticancer 2-arylpyridopyrimidinones that induce cytoplasmic and mitochondrial ROS and G2/M arrest.

Atherosclerosis, a leading cause of cardiovascular disease, is driven by the accumulation of oxidized low-density lipoproteins (oxLDL) in arterial walls. 7-Ketocholesterol (7KC), a major oxysterol found in oxLDL and atherosclerotic plaques, triggers multiple cell injuries including loss of lysosomal integrity, oxidative stress, apoptosis, and impaired autophagy in vascular cells. Bis(monoacylglycero)phosphate (BMP), also known as lysobisphosphatidic acid, is a unique phospholipid concentrated in the endolysosomal compartment, known to regulate vesicle dynamics, lysosomal enzyme activities, intracellular cholesterol trafficking and its oxidative metabolism. Using a validated model of BMP enrichment in murine RAW 264.7 macrophages, we investigated whether BMP could exert protective activity against 7KC-induced damage. Our findings revealed that BMP enrichment provides comprehensive protection against 7KC at the cellular level by preserving cell viability, morphology, and neutral lipid balance. Mechanistically, BMP enrichment prevented apoptosis by maintaining mitochondrial integrity and blocking caspase activation. This was demonstrated by normalized BAX/BCL2 ratios, preserved pro-Caspase-3 levels, and reduced PARP cleavage. Remarkably, BMP enrichment also restored autophagic flux, thereby preventing the pathological accumulation of LC3-II and p62 that characterizes autophagy dysfunction. Enhanced colocalization between LC3 and BMP suggests direct functional interactions in the stress response. Gene expression analysis confirmed that BMP enrichment normalized the transcriptional dysregulation of key autophagy regulators, including Sqstm1, Becn1, and Pink1. Taken together, these results suggest that BMP is an endogenous protective factor that counteracts 7KC-induced cellular damage at multiple steps by regulating cell death and autophagy pathways in a coordinated manner.