Cell Apoptotic Bodies Research Articles

Mechanism of autophagy inhibition for sensitizing HeLa cell apoptosis induced by trichosanthin

This research investigated the effect and mechanism of trichosanthin(TCS) in inducing autophagy and apoptosis of HeLa cells in cervical cancer. Two-step chromatography was used to prepare TCS. MTT assay was used to detect the inhibition effect of TCS on the proliferation of HeLa cells. The formation of autophagic vesicles and apoptotic bodies in HeLa cells treated with TCS was observed under transmission electron microscopy, and green fluorescent protein(GFP)-microtubule-associated protein 1 light chain beta 3(LC3B) fusion protein location was observed under laser confocal microscopy. The expression of autophagy and apoptosis-related proteins was detected by Western blot. It was found that the prepared TCS had high purity and biological activity. TCS could inhibit the proliferation of HeLa cells significantly in a concentration-dependent and time-dependent manner. Obvious autophagic vesicles were observed after HeLa cells were treated with TCS for 12 h, and typical apoptotic bodies were formed after 48 h. Under laser confocal microscopy, LC3B protein was observed to shift from a diffuse distribution to a spotted aggregation distribution in the cytoplasm. Western blot results showed that the expression of autophagy-activating protein LC3BⅡ began to increase in HeLa cells after being treated with TCS for 12 h and increased with the increase in drug concentration. Apoptosis-related protein poly ADP-ribose polymerase(PARP) began to activate as cleaved PARP after being treated with TCS for 24 h. The combination of TCS and 3-methyladenine(3-MA) further promoted the activation of cleaved PARP. The results showed that TCS could significantly inhibit the growth of HeLa cells in cervical cancer. Moreover, TCS induced autophagy in HeLa cells earlier than apoptosis, and inhibiting autophagy could sensitize apoptosis of HeLa cells induced by TCS. This indicated that autophagy induced by TCS was a protective cell response, and TCS combined with autophagy inhibitors could enhance the anti-cervical cancer effect of TCS.

An optically responsive cancer vaccine for inducing robust anti-tumor immunity by apoptotic body carrying nanoadjuvants

An optically responsive cancer vaccine for inducing robust anti-tumor immunity by apoptotic body carrying nanoadjuvants

The effectiveness of semi-wild Sumatran mango (Mangifera spp.) leaves as a phytotherapy agent for breast cancer

Background Breast cancer ranks first in the world, standing at a mortality rate of 24.5% per year and is the leading cause of cancer death in Indonesia. The current management of breast cancer therapy is considered less effective because of its careful use due to side effects that are detrimental to the patient. The semi-wild species from Sumatra are neglected and underutilized species but have the potential as a therapeutic agent. Previous research has revealed that this species of mango is high in antioxidant compounds. Objective This study was carried out to discover the anticancer activities of the semi-wild mango species via inhibitory activities and morphological changes in Michigan Cancer Foundation-7 (MCF-7) cells. Materials and methods The IC50 value of Mangifera sumatrana, Mangifera foetida, and Mangifera laurina leaves in n-hexane, ethyl-acetate, and methanol extracts was determined using the reagent 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Data were analyzed by two-way analysis of variance using IBM SPSS Statistics 21. Results and conclusion M. laurina n-hexane extracts exhibited anticancer activity (IC50 13.25 ppm). Nonpolar solutions were chosen as the most effective extraction solvent in anticancer tests because lipids in nonpolar solvents can hit the lipid bilayer. A hexane fraction was created by separating the majority of nonpolar fatty acid esters, and this fraction had a considerable impact on cytotoxic and apoptotic effects on MCF-7 cells. Therefore, all treatments can transform MCF-7 cells’ morphology into blackened dead cells that are degraded into small parts, such as apoptotic bodies in cells undergoing apoptotic processes. MTT assays against MCF-7 on three species of semi-wild Sumatran mango in different extraction solvents showed that n-hexane extracts of M. laurina had stronger anticancer activity than other samples. This study provides new information to support the development of standardized herbal medicines and phytopharmaca in the future.

Da-Chai-Hu-Tang Formula inhibits the progression and metastasis in HepG2 cells through modulation of the PI3K/AKT/STAT3-induced cell cycle arrest and apoptosis

Da-Chai-Hu-Tang Formula inhibits the progression and metastasis in HepG2 cells through modulation of the PI3K/AKT/STAT3-induced cell cycle arrest and apoptosis

Synthesis, characterization, DFT calculations, and anticancer properties of VI-period and f-block elements-substituted zinc oxide nanopowders

Synthesis, characterization, DFT calculations, and anticancer properties of VI-period and f-block elements-substituted zinc oxide nanopowders

Cobalt oxide nanoparticles functionalized with glucose and conjugated to Ellagic acid (Co3O4@Glu-Ellagic acid NPs) induce apoptotic genes in HepG2, liver cancer cell line

Cobalt oxide nanoparticles functionalized with glucose and conjugated to Ellagic acid (Co3O4@Glu-Ellagic acid NPs) induce apoptotic genes in HepG2, liver cancer cell line

Anti-hepatocellular carcinoma activity of Sorbaria sorbifolia by regulating VEGFR and c-Met/apoptotic pathway

Anti-hepatocellular carcinoma activity of Sorbaria sorbifolia by regulating VEGFR and c-Met/apoptotic pathway

In vitro profiling and molecular dynamics simulation studies of berberine loaded MCM-41 mesoporous silica nanoparticles to prevent neuronal apoptosis.

Neuronal loss in Alzheimer's disease has been reported to display features of apoptosis, pyroptosis (programmed necrosis), or necroptosis. This study thoroughly examines the production and characterization of MCM-41 based berberine (BBR)-loaded porous silica nanoparticles (MSNs) by a modified Stöber method, focusing on their possible role in inhibiting the apoptotic process. Particle size, polydispersity index, morphology, drug loading, zeta potential, entrapment efficiency, and drug release were examined. The formulation was analyzed using various spectroscopic techniques. The surface area was computed by the Brunauer-Emmett-Teller plot. Computational models were developed for molecular dynamics simulation studies. A small PDI value indicated an even distribution of particles at nanoscale sizes (80-100 nm). Results from XRD and SEAD experiments confirmed the amorphous nature of BBR in nanoparticles. Nanoparticles had high entrapment (75.21 ± 1.55%) and drug loading (28.16 ± 2.5%) efficiencies. A negative zeta potential value (-36.861.1 mV) indicates the presence of silanol groups on the surface of silica. AFM findings reveal bumps due to the surface drug that contributed to the improved roughness of the MSNs-BBR surface. Thermal gravimetric analysis confirmed the presence of BBR in MSNs. Drug release was controlled by simple diffusion or quasi-diffusion. Molecular dynamics simulations confirmed the existence of diffused drug molecules. Cellular studies using SH-SY-5Y cells revealed dose-dependent growth inhibition. Fragmented cell nuclei and nuclear apoptotic bodies in DAPI-stained cells exposed to nanoparticles showed an increase in apoptotic cells. Flow cytometry analysis demonstrated a lower red-to-green ratio in SH-SY-5Y cells treated with nanoparticles. This suggests improved mitochondrial health, cellular viability restoration, and prevention of the apoptotic process. This study provides essential data on the synthesis and potential of MSNs loaded with BBR, which may serve as a viable therapeutic intervention for conditions associated with apoptosis.

A second-generation M1-polarized CAR macrophage with antitumor efficacy.

Chimeric antigen receptor (CAR) T cell therapies have successfully treated hematological malignancies. Macrophages have also gained attention as an immunotherapy owing to their immunomodulatory capacity and ability to infiltrate solid tumors and phagocytize tumor cells. The first-generation CD3ζ-based CAR-macrophages could phagocytose tumor cells in an antigen-dependent manner. Here we engineered induced pluripotent stem cell-derived macrophages (iMACs) with toll-like receptor 4 intracellular toll/IL-1R (TIR) domain-containing CARs resulting in a markedly enhanced antitumor effect over first-generation CAR-macrophages. Moreover, the design of a tandem CD3ζ-TIR dual signaling CAR endows iMACs with both target engulfment capacity and antigen-dependent M1 polarization and M2 resistance in a nuclear factor kappa B (NF-κB)-dependent manner, as well as the capacity to modulate the tumor microenvironment. We also outline a mechanism of tumor cell elimination by CAR-induced efferocytosis against tumor cell apoptotic bodies. Taken together, we provide a second-generation CAR-iMAC with an ability for orthogonal phagocytosis and polarization and superior antitumor functions in treating solid tumors relative to first-generation CAR-macrophages.

Potent anti-cancer activity of Sphaerocoryne affinis fruit against cervical cancer HeLa cells via inhibition of cell proliferation and induction of apoptosis

BackgroundCervical cancer remains a significant global health issue, highlighting the need for effective therapeutic strategies. Given that Sphaerocoryne affinis (SA) has shown potential anti-cancer activity in several cancer types, herein, we investigate the effects of SA fruit (SAF) on human cervical cancer HeLa cells and their underlying mechanisms of action.MethodsSAF extract cytotoxicity was assessed in various cancer cell lines. The effects of the hexane fraction (SAF-Hex) on HeLa cell viability, cell cycle protein expression, apoptosis, and DNA damage were evaluated using cytotoxicity assays, Western blotting, quantitative PCR, 4′,6-diamidino-2-phenylindole (DAPI) staining, and a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.ResultsSAF-Hex selectively inhibited HeLa cell viability with an IC50 of 4.20 ± 0.36 µg/mL and a selectivity index of 5.11 ± 0.58. The time-dependent cytotoxicity assay showed decreased cell survival after 48 h of treatment, accompanied by morphological changes and apoptotic bodies in HeLa cells. SAF-Hex also suppressed HeLa cell cycle proteins (Cyclin E, CDK2, and CDK1), reduced PCNA transcription, and diminished AKT and mTOR activation, thus inhibiting cell proliferation. The increased γH2AX expression, DNA fragmentation, and caspases-3 and -9 activation indicated SAF-Hex-induced DNA damage and apoptosis. However, the BAX/BCL-2 ratio remained unchanged, and BAX and BCL2 expression was attenuated.ConclusionSAF-Hex effectively inhibits HeLa cell proliferation and induces DNA damage in that cervical cancer cell line activating apoptosis through the intrinsic pathway. Interestingly, the BAX/BCL-2 ratio remained unchanged while BAX and BCL2 transcription was attenuated. Hence, further research is required to explore this unexpected finding and facilitate the development of novel therapies targeting cervical cancer HeLa cells.

Oenothein B inhibits proliferation and migration of breast cancer cells by regulating P53

The effects of oenothein B(OEB) on the proliferation, apoptosis, invasion, and migration of breast cancer MCF-7 and MDA-MB-231 cells were investigated by cell culture in vitro, network pharmacology, and molecular docking. In vitro cell experiments revealed that OEB inhibited the proliferation and colony formation ability, and promoted the apoptosis and formation of apoptotic bodies in breast cancer cells, as well as inhibited the invasion and migration of breast cancer cells. The targets of OEB were obtained using SwissTargetPrediction database and breast cancer targets were obtained from GeneCards. The targets of OEB and breast cancer were entered separately in Venny 2.1 software to obtain the Venn diagram of common targets of OEB and breast cancer. The common targets of OEB and breast cancer were input into STRING database to construct a protein-protein interaction(PPI) network, which was entered into Cytoscape 3.7.2 software for network topology analysis. Key targets were screened according to protein association strength, and analyzed for KEGG pathway enrichment. Molecular docking of OEB to key targets using AutoDock software revealed that OEB stably bound to the active pocket of P53, while OEB promoted the expression of P53 protein. MCF-7 and MDA-MB-231 cell viability and migration ability increased after silencing P53, and this change was reversed after treatment with OEB. Therefore, this study showed that OEB inhibited the proliferation, migration, and invasion of breast cancer MCF-7 and MDA-MB-231 cells, and promoted the apoptosis of breast cancer MCF-7 and MDA-MB-231 cells, which may be related to the targeted regulation of P53.

Exploring the Metabolic Differences between Cisplatin- and UV Light-Induced Apoptotic Bodies in HK-2 Cells by an Untargeted Metabolomics Approach.

Among the extracellular vesicles, apoptotic bodies (ABs) are only formed during the apoptosis and perform a relevant role in the pathogenesis of different diseases. Recently, it has been demonstrated that ABs from human renal proximal tubular HK-2 cells, either induced by cisplatin or by UV light, can lead to further apoptotic death in naïve HK-2 cells. Thus, the aim of this work was to carry out a non-targeted metabolomic approach to study if the apoptotic stimulus (cisplatin or UV light) affects in a different way the metabolites involved in the propagation of apoptosis. Both ABs and their extracellular fluid were analyzed using a reverse-phase liquid chromatography-mass spectrometry setup. Principal components analysis showed a tight clustering of each experimental group and partial least square discriminant analysis was used to assess the metabolic differences existing between these groups. Considering the variable importance in the projection values, molecular features were selected and some of them could be identified either unequivocally or tentatively. The resulting pathways indicated that there are significant, stimulus-specific differences in metabolites abundancies that may propagate apoptosis to healthy proximal tubular cells; thus, we hypothesize that the share in apoptosis of these metabolites might vary depending on the apoptotic stimulus.

Large-scale morphometry of the subarachnoid space of the optic nerve

BackgroundThe meninges, formed by dura, arachnoid and pia mater, cover the central nervous system and provide important barrier functions. Located between arachnoid and pia mater, the cerebrospinal fluid (CSF)-filled subarachnoid space (SAS) features a variety of trabeculae, septae and pillars. Like the arachnoid and the pia mater, these structures are covered with leptomeningeal or meningothelial cells (MECs) that form a barrier between CSF and the parenchyma of the optic nerve (ON). MECs contribute to the CSF proteome through extensive protein secretion. In vitro, they were shown to phagocytose potentially toxic proteins, such as α-synuclein and amyloid beta, as well as apoptotic cell bodies. They therefore may contribute to CSF homeostasis in the SAS as a functional exchange surface. Determining the total area of the SAS covered by these cells that are in direct contact with CSF is thus important for estimating their potential contribution to CSF homeostasis.MethodsUsing synchrotron radiation-based micro-computed tomography (SRµCT), two 0.75 mm-thick sections of a human optic nerve were acquired at a resolution of 0.325 µm/pixel, producing images of multiple terabytes capturing the geometrical details of the CSF space. Special-purpose supercomputing techniques were employed to obtain a pixel-accurate morphometric description of the trabeculae and estimate internal volume and surface area of the ON SAS.ResultsIn the bulbar segment, the ON SAS microstructure is shown to amplify the MECs surface area up to 4.85-fold compared to an “empty” ON SAS, while just occupying 35% of the volume. In the intraorbital segment, the microstructure occupies 35% of the volume and amplifies the ON SAS area 3.24-fold.ConclusionsWe provided for the first time an estimation of the interface area between CSF and MECs. This area is of importance for estimating a potential contribution of MECs on CSF homeostasis.

Exosome transportation-mediated immunosuppression relief through cascade amplification for enhanced apoptotic body vaccination

Exosome transportation-mediated immunosuppression relief through cascade amplification for enhanced apoptotic body vaccination

Engulf and digest – TRPM7 as key regulator of macrophage phagosome maturation

Engulf and digest – TRPM7 as key regulator of macrophage phagosome maturation

Propofol and Sevoflurane Alleviate Malignant Biological Behavior and Cisplatin Resistance of Xuanwei Lung Adenocarcinoma by Modulating the Wnt/β-catenin Pathway and PI3K/AKT Pathway.

Recent studies have shown that propofol and sevoflurane, two common anesthetics, can prevent tumor development. The prevalence of lung adenocarcinoma in China is highest in Yunnan Xuanwei, and many patients with lung cancer in this area are often resistant to platinum-based treatments. The objective of the study was to investigate the effects of propofol and sevoflurane on malignant biological behavior and cisplatin resistance of Xuanwei lung adenocarcinoma. Herein, XWLC-05/R, a cisplatin-resistant cell line of XWLC-05 cells from Xuanwei lung adenocarcinoma, was constructed. The XWLC-05 cells and XWLC-05/R cells were treated with propofol and sevoflurane singly or as a combination and subjected to CCK-8 assay, clone formation tests, and flow cytometry analysis to assess the proliferation level of cells. The morphology and number of apoptotic bodies in XWLC-05 cells and XWLC-05/R were examined with a transmission electron microscope. The ANNEXIN V-FITC/PI and transwell assays were performed to evaluate apoptosis, invasion, and migration of the cells. Subsequently, we constructed a Xuanwei lung adenocarcinoma xenograft model to investigate the effects of propofol and sevoflurane on the tumorigenicity of XWLC-05 cells in vivo. Treatment with propofol and sevoflurane significantly inhibited proliferation, invasion, migration, and induced apoptosis of XWLC-05 and XWLC-05/R cells. These effects were more pronounced when propofol and sevoflurane were co-incubated with the cells. Moreover, both propofol and sevoflurane significantly inhibited Wnt/β- catenin and PI3K/AKT pathways. Moreover, the two drugs effects suppressed the malignant biological behavior of XWLC-05 cells and XWLC-05/R cells, and this effect was counteracted by 740Y-P (PI3K/AKT pathway activator) and Wnt pathway activator 1 (Wnt/β-catenin pathway activator). In vivo experiments confirmed that propofol and sevoflurane alleviated the tumorigenicity of XWLC-05 cells. In summary, this study shows, for the first time, that propofol and sevoflurane decrease the proliferation, invasion, migration and induce apoptosis of XWLC-05 cells and XWLC-05/R cells by impeding the Wnt/β-catenin and PI3K/AKT signaling pathways, thereby alleviating the development of Xuanwei lung adenocarcinoma in vivo. Moreover, these effects are more pronounced when the two drugs are combined.

Abstract 1329: Maresins prevent breast cancer dormancy escape via resolution of inflammation

Abstract Cytotoxic cancer therapies reduce tumor burden by killing tumor cells. However, the resulting apoptotic and necrotic cell bodies (tumor cell “debris”) may stimulate tumor initiation and progression by disrupting the resolution of inflammation. Thus, chemotherapy and anti-estrogen breast cancer therapy, including tamoxifen, may be a double-edged sword. A paradigm shift is emerging in understanding the resolution of inflammation as an active biochemical process with the discovery of novel specialized pro-resolving lipid autocoid mediators (SPMs), such as maresins and endogenous resolution programs. Despite approaches to block systemic inflammation, there are no current “pro-resolving” therapies in cancer. To determine whether debris stimulates breast cancer growth, we utilized tumor dormancy models with a subthreshold (nontumorigenic) inoculum of tumor cells. We demonstrated that breast tumor “debris” generated by cytotoxic anti-estrogen therapy (tamoxifen or fulvestrant) or chemotherapy (eribulin) stimulates dormancy escape by triggering a macrophage-derived pro-inflammatory and pro-angiogenic “cytokine storm”. Thus, tumor cell debris is a critical pro-tumorigenic factor in breast cancer initiation and progression. To assess whether stimulating the clearance of debris would suppress breast cancer progression, we utilized the SPMs maresin 1 (MaR1) and maresin conjugates in tissue regeneration (MCTR1, MCTR2). Each maresin (MaR1, MCTR1 and MCTR2) sharply reduced tumor growth in both debris-stimulated and spontaneous (e.g. MMTV-PyMT) breast cancer models at nanogram concentrations (15 ng/day) without toxicity. Notably, maresins enhanced immunotherapy (anti-CTLA4) to induce tumor regression in estrogen receptor (ER) positive (EO771) and inhibit ER negative tumor growth (4T1). Maresins stimulated macrophage phagocytosis of therapy (fulvestrant and tamoxifen)-generated breast cancer debris at only nanomolar concentrations (0.1 - 10 nM). Remarkably, maresins alone or in combination with chemotherapy (paclitaxel) reduced levels of pro-angiogenic factors (e.g. CXCL12/SDF-1) in the tumor microenvironment and decreased microvessel density/size, thereby inhibiting tumor angiogenesis. Maresins dampened the therapy-induced cytokine storm, by reducing levels of TNF-α, MIP-2/CXCL2, CCL2/MCP-1, IL-1ra/IL-1F3, CCL5, CXCL13, Serpin E1/PAI-1, IL-1β and G-CSF both in vitro in debris-stimulated macrophages and in vivo in plasma and tumor tissue. Stimulating the resolution of inflammation via pro-resolution lipid mediators to enhance immunotherapy is a novel host-centric therapeutic approach to prevent breast cancer initiation, dormancy escape and tumor progression via debris clearance and counter-regulation of the cytokine storm. Altogether, the maresin pathway mediators may represent a new therapeutic approach to stimulate the resolution of inflammation in breast cancer. Citation Format: Franciele Cristina Kipper, Jianjun Deng, Eva Rothenberger, Abigail Kelly, Madeline Duncan, Sui Huang, Charles N. Serhan, Dipak Panigrahy. Maresins prevent breast cancer dormancy escape via resolution of inflammation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1329.

Effect of 1,4-Napthoquinone Derivatives on Anti-proliferation and Apoptosis Induction in Skin and Lung Cancer Cells

Background: Cancer is a major public health concern around the world. Nowadays, standard therapy in combination with targeted therapy are being used for cancer treatment, but these therapeutics show side effects or unwanted symptoms and high cost. 1,4-Napthoquinone and their derivatives were used in cancer research for a long time with high efficiency than some cancer drug. In the present study, the effect of a novel 1,4-napthoquinone, DL667 and DL666 on apoptosis induction in malignant melanoma, A375 cells and lung cancer, A549 cells, respectively were studied. Objective: To investigate the effects of 1,4-napthoquinone derivatives, DL667 and DL66, on apoptosis induction in malignant melanoma, A375 and lung cancer, A549 cells, respectively. Materials and Methods: Cell growth inhibition was determined by MTT assay. Apoptosis induction was via chromatin condensation or apoptotic bodies determined by Hoechst staining. In addition, changes in mitochondrial membrane potential (ΔΨm) were done by JC-1 staining. Results: The results showed that DL667 inhibited cell growth in A375 with an IC50 at 11.2+0.419 μM in a concentration-dependent manner and DL666 inhibited cell growth of A549 cells at 200 μM. Hoechst staining assay revealed that both DL667 and DL666 could induce chromatin condensation and apoptotic bodies in A375 and A549 cells. In addition, the JC-1 staining showed that DL667 and DL666 decreased the ΔΨm, a marker event of early apoptosis via mitochondria, in A375 and A549 treated cells. Conclusion: DL667 and DL666 showed anti-proliferation and apoptosis induction in A375 and A549 cells, respectively. Moreover, DL667 and DL666 induced chromatin condensation, apoptotic bodies and decreased the ΔΨm, resulting in apoptosis cell death induction. Our results suggested that the decrease of ΔΨm may be further developed as a model for study in other cancer cell lines. However, the underlying mechanisms of apoptosis induction associated with signaling should be further studied. Keywords: 1,4-Napthoquinone; Apoptosis; Chromatin condensation; Apoptotic bodies; Mitochondrial membrane potential

Preparation and Characterization of Novel Schiff Base Derived From 4-Nitro Benzaldehyde and Its Cytotoxic Activities.

Normal drugs exhibit activities against both normal and cancer cells. Furthermore, cancer cells may develop resistance to these drugs that alternative treatment must be explored. The main objective of this study was to examine the anticancer activity of Schiff base against Tongue Squamous Cell Carcinoma Fibroblasts (TSCCF) and normal human gingival fibroblasts (NHGF) and to propose its mechanism. A Novel Schiff base ligand was synthesized from the reaction of 5-C-2-4-NABA (5-chloro-2-((4-nitrobenzylidene) amino) benzoic acid). These Schiff bases possessed azomethine group (-HC=N-) and aromatic group (CH) as analyzed by Fourier transforms infrared (FTIR) spectroscopy and UV-Vis spectra. The in vitro cytotoxicity screening assay suggested promising activity against TSCCF with IC50 of 446.68 µg/mL, but insignificant activity against NHGF cells (IC50 of 977.24 µg/mL) after 72 h. The evidence of apoptotic induction was supported by DAPI staining of apoptotic nuclei with reduced cell numbers, suggesting that Schiff base could induce apoptotic bodies in cancer cells being observed. Based on the Schiff base structure, the anti-cancer mechanism may be attributed to the -HC=N- azomethine group. For the first time, our findings highlighted the anticancer activities of the new Schiff base against oral cancer cell lines.

Titaniumdioxide mediated sonophotodynamic therapy against prostate cancer.

Titaniumdioxide mediated sonophotodynamic therapy against prostate cancer.