CASP8 Genes Research Articles

Impacts of Bacteriostatic and Bactericidal Antibiotics on the Mitochondria of the Age-Related Macular Degeneration Cybrid Cell Lines.

We assessed the potential negative effects of bacteriostatic and bactericidal antibiotics on the AMD cybrid cell lines (K, U and J haplogroups). AMD cybrid cells were created and cultured in 96-well plates and treated with tetracycline (TETRA) and ciprofloxacin (CPFX) for 24 h. Reactive oxygen species (ROS) levels, mitochondrial membrane potential (ΔψM), cellular metabolism and ratio of apoptotic cells were measured using H2DCFDA, JC1, MTT and flow cytometry assays, respectively. Expression of genes of antioxidant enzymes, and pro-inflammatory and pro-apoptotic pathways were evaluated by quantitative real-time PCR (qRT-PCR). Higher ROS levels were found in U haplogroup cybrids when treated with CPFX 60 µg/mL concentrations, lower ΔψM of all haplogroups by CPFX 120 µg/mL, diminished cellular metabolism in all cybrids with CPFX 120 µg/mL, and higher ratio of dead cells in K and J cybrids. CPFX 120 µg/mL induced overexpression of IL-33, CASP-3 and CASP-9 in all cybrids, upregulation of TGF-β1 and SOD2 in U and J cybrids, respectively, along with decreased expression of IL-6 in J cybrids. TETRA 120 µg/mL induced decreased ROS levels in U and J cybrids, increased cellular metabolism of treated U cybrids, higher ratio of dead cells in K and J cybrids and declined ΔψM via all TETRA concentrations in all haplogroups. TETRA 120 µg/mL caused upregulation of IL-6 and CASP-3 genes in all cybrids, higher CASP-7 gene expression in K and U cybrids and downregulation of the SOD3 gene in K and U cybrids. Clinically relevant dosages of ciprofloxacin and tetracycline have potential adverse impacts on AMD cybrids possessing K, J and U mtDNA haplogroups in vitro.

Oxidative stress, apoptosis and serotonergic system changes in zebrafish (Danio rerio) gills after long-term exposure to microplastics and copper

Fish gills are in direct contact with the surrounding pollutants, and thus, potentially more vulnerable to microplastics (MPs) and heavy metals. The present study aimed to evaluate the long-term exposure effects of MPs and copper (Cu) in the gills of adult zebrafish (Danio rerio). To this end, zebrafish were exposed to MPs (2 mg/L), Cu (Cu25, 25 μg/L) and their mixture (Cu25 + MPs) for 30 days, and then oxidative stress, detoxification, antioxidant, metabolic and neurotoxicity enzymes/genes, as well serotonergic system and apoptosis genes, were evaluated in gills. In the mixture group, ROS levels were increased, while CAT and GPx activities were inhibited, indicating the induction of oxidative stress in zebrafish gills. This was followed by an increase of LPO levels and potential oxidative damage in zebrafish gills. The tryptophan hydroxylase 1a (tph1a) and caspase-3 (casp3) genes were significantly upregulated in Cu25 + MPs group, indicating a potential dysregulation of serotonin synthesis and apoptosis pathways, respectively. Overall, the present study contributes to improving the knowledge about the response of aquatic organisms to MPs and the potential ecological risk that these particles represent to the ecosystems.

Integrative analysis of genomic and transcriptomic data of normal, tumour, and co-occurring leukoplakia tissue triads drawn from patients with gingivobuccal oral cancer identifies signatures of tumour initiation and progression.

A thickened, white patch — leukoplakia — in the oral cavity is usually benign, but sometimes (in ~9% of individuals) it progresses to malignant tumour. Because the genomic basis of this progression is poorly understood, we undertook this study and collected samples of four tissues — leukoplakia, tumour, adjacent normal, and blood — from each of 28 patients suffering from gingivobuccal oral cancer. We performed multiomics analysis of the 112 collected tissues (four tissues per patient from 28 patients) and integrated information on progressive changes in the mutational and transcriptional profiles of each patient to create this genomic narrative. Additionally, we generated and analysed whole‐exome sequence data from leukoplakia tissues collected from 11 individuals not suffering from oral cancer. Nonsynonymous somatic mutations in the CASP8 gene were identified as the likely events to initiate malignant transformation, since these were frequently shared between tumour and co‐occurring leukoplakia. CASP8 alterations were also shown to enhance expressions of genes that favour lateral spread of mutant cells. During malignant transformation, additional pathogenic mutations are acquired in key genes (TP53, NOTCH1, HRAS) (41% of patients); chromosomal‐instability (arm‐level deletions of 19p and q, focal‐deletion of DNA‐repair pathway genes and NOTCH1, amplification of EGFR) (77%), and increased APOBEC‐activity (23%) are also observed. These additional alterations were present singly (18% of patients) or in combination (68%). Some of these alterations likely impact immune‐dynamics of the evolving transformed tissue; progression to malignancy is associated with immune suppression through infiltration of regulatory T‐cells (56%), depletion of cytotoxic T‐cells (68%), and antigen‐presenting dendritic cells (72%), with a concomitant increase in inflammation (92%). Patients can be grouped into three clusters by the estimated time to development of cancer from precancer by acquiring additional mutations (range: 4–10 years). Our findings provide deep molecular insights into the evolutionary processes and trajectories of oral cancer initiation and progression. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Exposure to a mixture of heavy metals induces cognitive impairment: Genes and microRNAs involved

Converging evidence demonstrates that microRNAs (miRNAs) play an important role in the etiology of cognitive impairment. Thus, we aim to: (i) identify the molecular mechanisms of heavy metals, particularly miRNAs involved in the development of cognitive impairment; and (ii) generate miRNA sponges to prevent them from binding with their target messenger RNAs. The Comparative Toxicogenomics Database (CTD; http://ctd.mdibl.org), MicroRNA ENrichment TURned NETwork (MIENTURNET, http://userver.bio.uniroma1.it/apps/mienturnet/) and the microRNA sponge generator and tester (miRNAsong, http://www.med.muni.cz/histology/miRNAsong) were used as the core data-mining approaches in the current study. We observed that lead acetate, arsenic, gold, copper, iron, and aluminum, as well as their mixtures, had significant effects on the development of cognitive impairment. Although prevalent genes obtained from investigated heavy metals of cognitive impairment were different, the "PI3K-Akt signaling pathway", "pathways of neurodegeneration-multiple diseases", "apoptosis", "apoptosis-multiple species", "p53 signaling pathway", "NF-kappa B signaling pathway", and "Alzheimer’s disease pathway" were highlighted. The mixed heavy metals altered the genes BAX, CASP3, BCL2, TNF, and IL-1B, indicating the significance of apoptosis and pro-inflammatory cytokines in the pathogenesis of cognitive impairment and the possibility of targeting these genes in future neuroprotective therapy. In addition, we used a network-based approach to identify key genes, miRNAs, pathways, and diseases related to the development of cognitive impairment. We also found 16 significant miRNAs related to cognitive impairment (hsa-miR-1–3p, hsa-let-7a-5p, hsa-miR-9–5p, hsa-miR-16–5p, hsa-miR-17–5p, hsa-miR-20a-5p, hsa-miR-26a-5p, hsa-miR-26b-5p, hsa-miR-34a-5p, hsa-miR-101–3p, hsa-miR-106a-5p, hsa-miR-128–3p, hsa-miR-144–3p, hsa-miR-199a-3p, hsa-miR-204–5p, and hsa-miR-335–5p). Finally, we created and evaluated miRNA sponge sequences for these miRNAs in silico. Further studies, including in vivo and in vitro, are needed to assess the link between these genes, miRNAs, pathways, and cognitive impairment.

Study on the Mechanism of Astragalus Polysaccharide in Treating Pulmonary Fibrosis Based on "Drug-Target-Pathway" Network.

Pulmonary fibrosis is a chronic, progressive and irreversible heterogeneous disease of pulmonary interstitial tissue. Its incidence is increasing year by year in the world, and it will be further increased due to the pandemic of COVID-19. However, at present, there is no safe and effective treatment for this disease, so it is very meaningful to find drugs with high efficiency and less adverse reactions. The natural astragalus polysaccharide has the pharmacological effect of anti-pulmonary fibrosis with little toxic and side effects. At present, the mechanism of anti-pulmonary fibrosis of astragalus polysaccharide is not clear. Based on the network pharmacology and molecular docking method, this study analyzes the mechanism of Astragalus polysaccharides in treating pulmonary fibrosis, which provides a theoretical basis for its further clinical application. The active components of Astragalus polysaccharides were screened out by Swisstarget database, and the related targets of pulmonary fibrosis were screened out by GeneCards database. Protein-protein interaction network analysis and molecular docking were carried out to verify the docking affinity of active ingredients. At present, through screening, we have obtained 92 potential targets of Astragalus polysaccharides for treating pulmonary fibrosis, including 11 core targets. Astragalus polysaccharides has the characteristics of multi-targets and multi-pathways, and its mechanism of action may be through regulating the expression of VCAM1, RELA, CDK2, JUN, CDK1, HSP90AA1, NOS2, SOD1, CASP3, AHSA1, PTGER3 and other genes during the development of pulmonary fibrosis.

In vitro investigation of cytotoxic, antioxidant, apoptosis-inducing, and wound healing properties of endemic Centaurea fenzlii Reichardt extracts.

Cancer is a formidable problem for global health, and the increasing burden necessitates the search for new and alternative treatments and/or approaches. For this reason, any approach to cancer treatment is extremely valuable. Recently, there has been increasing interest in the use of plant-derived compounds in the treatment of cancer. ​Many studies have shown that many of the best-selling anti-cancer drugs are of plant origin. Based on this, the cytotoxic potential of two extracts [C. fenzlii leaf extract (CFL) and C. fenzlii capitula extract (CFC)] from Centaurea fenzlii, a plant species endemic to Turkey, were tested in two different cancer cell lines (DLD1 and ARH77) and a healthy cell line (HUVEC). Cytotoxic effects of the extracts were determined by MTT assay, changes in the mRNA expressions of the apoptotic genes (apaf, bax, bcl2, bcl2l11, casp3, gadd45a and hrk) were determined via the qRT-PCR technique, and the caspase3 enzyme activity was determined. In addition, the capacities of the extracts against invasion and migration were also evaluated. The chemical content and phenolic composition were evaluated via the gas chromatography-mass spectrometry (GC/MS) and high-performance liquid chromatography (HPLC) techniques. As a result, the extracts showed a selective cytotoxic effect in all the cancer cell lines examined, especially the DLD1 colorectal cell line. In addition, it has been determined that the extracts inhibit the invasion and migration of cells. The results of the study reveal that C. fenzlii, which has been found to have acceptable anti-cancer effects, should be investigated with more comprehensive studies.

Bioinformatic identification of differentially expressed genes regulated by DNA-methylation in glioblastoma.

DNA methylation-driven differentially expressed genes (DEGs) play potentially important roles in glioblastoma (GBM). In the present study, we applied bioinformatic analyses to identify key methylation-regulated DEGs (MeDEGs) in glioblastoma and elucidate their functions. Gene expression and methylation profile data from glioblastoma samples along with clinical information were obtained from GEO and TCGA databases. A total of 65 genes were identified as MeDEGs from the aforementioned data. Subsequently, gene ontology and kyoto encyclopaedia of genes and genomes enrichment analyses of these MeDEGs exhibited that MeDEGs were mostly enriched in several tumour-related terms such as 'activation of cysteine-type endopeptidase activity involved in apoptotic process' and 'phospholipid scrambling'. Kaplan-Meier survival analysis demonstrated significant correlation of CASP1, CFH and TTLL7 hyper-methylation with patient prognosis. Finally, CASP1 protein could indirectly interact with CFH protein, but interaction of TTLL7 protein with CASP1 or CFH protein was not evident. Based on gene set enrichment analysis, hyper-methylation of CASP1, CFH and TTLL7 were found enriched in tumour-related KEGG terms, such as 'RNA degradation', 'apyruvate metabolism' and 'nitrogen metabolism'. Methylation levels of CASP1, CFH and TTLL7 were addressed to negatively correlate with their mRNA levels in GBM cell lines. In sum, the present identification of MeDEGs associated with overall survival put forth potential molecular targets for translation towards improved diagnosis and treatment of GBM, and specifically, methylation levels of CASP1, CFH and TTLL7 genes could serve as key prognostic biomarkers in GBM.

Comparative assessment on the probable mechanisms underlying the hepatorenal toxicity of commercial imidacloprid and hexaflumuron formulations in rats

Pesticides are viewed as a major wellspring of ecological contamination and causing serious risky consequences for people and animals. Imidacloprid (IM) and hexaflumuron (HFM) are extensively utilized insect poisons for crop assurance on the planet. A few investigations examined IM harmfulness in rodents, but its exact mechanism hasn’t been mentioned previously as well as the toxicity of HFM doesn’t elucidate yet. For this reason, the present study was designed to explore the mechanism of each IM and HFM–evoked rat liver and kidney toxicity and to understand its molecular mechanism. 21 male Wistar albino rats were divided into 3 groups, as follows: group (1), normal saline; group (2), IM; and group (3), HFM. Both insecticides were orally administered every day for 28 days at a dose equal to 1/10 LD50 from the active ingredient. After 28 days postdosing, rats were anesthetized to collect blood samples then euthanized to collect liver and kidney tissue specimens. The results showed marked changes in walking, body tension, alertness, and head movement with a significant reduction in rats’ body weight in both IM and HFM receiving groups. Significant increases in MDA levels and decrease of GHS levels were recorded in liver and kidney homogenates of either IM or HFM groups. Liver and kidney tissues obtained from both pesticide receiving groups showed extensive histopathological alterations with a significant increase in the serum levels of ALT, AST, urea, and creatinine and a decrease in total proteins, albumin, and globulin levels. In addition, there was upregulation of the transcript levels of casp-3, JNK, and HO-1 genes with strong immunopositivity of casp-3, TNF-ὰ, and NF-KB protein expressions in the liver and kidneys of rats receiving either IM or HFM compared with the control group. In all studied parameters, HFM caused hepatorenal toxicity more than those induced by IM. We can conclude that each IM and HFM provoked liver and kidneys damage through overproduction of ROS, activation of NF-KB signaling pathways and mitochondrial/JNK-dependent apoptosis pathway.

Environmental relevant concentrations of triclosan affected developmental toxicity, oxidative stress, and apoptosis in zebrafish embryos.

Triclosan (TCS), as a broad-spectrum antibacterial agent, is widely used in various pharmaceutical and personal care products. However, the details of ecological environmental health risks of TCS are not clear. In this study, zebrafish embryos/larval were exposed to environmentally relevant concentrations of TCS to evaluate the developmental toxicity. Four-hour post-fertilization (hpf) zebrafish embryos were exposed to 0, 2, 10, 50, and 250μg/L TCS until 96 h. The heart beats at 72 hpf were significantly increased in 2μg/L TCS group, while significantly decreased in 250μg/L TCS treated group compared with control. The results of acridine orange staining, terminal deoxynucleotide transferase-mediated UTPnick end labeling assay, and detection of mitochondrial membrane potential showed that 50 and 250μg/L TCS resulted in apoptosis. Meanwhile, reactive oxygen species (ROS) and DNA damage were induced, but SOD activity was significantly decreased in 250μg/L TCS treated group. In addition, SOD(Mn) and GPx gene mRNA expressions were significantly down-regulated in 50 and 250μg/L TCS treated groups, while Casp3, Casp9, Puma, Casp8, Apaf1, and Bid genes in 250μg/L TCS and Mdm2 gene in 50μg/L treated groups were significantly up-regulated. P53 protein was significantly up-regulated in 250μg/L TCS treated group. The overall results showed that TCS can cause oxidative stress and result in apoptosis via the involvement of ROS-p53-caspase-dependent apoptotic pathway in zebrafish embryos. The present findings suggest the potential mechanisms of TCS-induced developmental toxicity appears to be the generation of ROS and the consequent triggering of apoptosis genes.

Growth Inhibition and Apoptotic Effect of Pine Extract and Abietic Acid on MCF-7 Breast Cancer Cells via Alteration of Multiple Gene Expressions Using In Vitro Approach.

In vitro anti-proliferative activity of Pinus palustris extract and its purified abietic acid was assessed against different human cancer cell lines (HepG-2, MCF-7 and HCT-116) compared to normal WI-38 cell line. Abietic acid showed more promising IC50 values against MCF-7 cells than pine extract (0.06 µg/mL and 0.11 µM, respectively), with insignificant cytotoxicity toward normal fibroblast WI-38 cells. Abietic acid triggered both G2/M cell arrest and subG0-G1 subpopulation in MCF-7, compared to SubG0-G1 subpopulation arrest only for the extract. It also induced overexpression of key apoptotic genes (Fas, FasL, Casp3, Casp8, Cyt-C and Bax) and downregulation of both proliferation (VEGF, IGFR1, TGF-β) and oncogenic (C-myc and NF-κB) genes. Additionally, abietic acid induced overexpression of cytochrome-C protein. Furthermore, it increased levels of total antioxidants to diminish carcinogenesis and chemotherapy resistance. P. palustris is a valuable source of active abietic acid, an antiproliferative agent to MCF-7 cells through induction of apoptosis with promising future anticancer agency in breast cancer therapy.

Protein-coding gene interaction network prediction of bioactive plant compound action against SARS-CoV-2: a novel hypothesis using bioinformatics analysis.

This study aimed to verify the action of bioactive compounds from Brazilian plants on the leader genes involved in the SARS-CoV-2 pathway. The main human genes involved were identified in GeneCards and UNIPROT platforms, and an interaction network between leader genes was established in the STRING database. To design chemo-biology interactome networks and elucidate the interplay between genes related to the disease and bioactive plant compounds, the metasearch engine STITCH 3.1 was used. The analysis revealed that SMAD3 and CASP3 genes are leader genes, suggesting that the mechanism of action of the virus on host cells is associated with the molecular effects of these genes. Furthermore, the bioactive plant compounds, such as ascorbate, benzoquinone, ellagic acid, and resveratrol was identified as a promising adjuvant for the treatment inhibiting CASP3-mediated apoptosis. Bioactive plant compounds were verified as the main pathways enriched with KEGG and related to viral infection, assessments/immune/infections, and cell proliferation, which are potentially used for respiratory viral infections. The best-ranked molecule docked in the CASP3 binding site was rutin, while the SMAD3 binding site was resveratrol. In conclusion, this work identified several bioactive compounds from Brazilian plants showing potential antiviral functions that can directly or indirectly inhibit the new coronavirus.

Impact assessment of cadmium chloride on human cell lines and yeast knockout strains

Background Cadmium (Cd) is a heavy metal that contributes to pollution in the environment. Cd intoxication can lead to diseases in the liver, kidneys, and lungs, among other organs. The liver is the primary organ affected by Cd overdose.Objective To evaluate the cytotoxic and genotoxic responses of cadmium chloride (CdCl2) on three different human cell lines and four different genotypes of yeast knockout strains.Materials and methods The effects of different concentrations of CdCl2 on cell viability in hepatocellular carcinoma (HepG2), lung cancer (A549), and normal lung cell carcinoma (Wi38) were investigated systematically using the MTT method. Moreover, flow cytometry was used to assess cell cycle arrest using propidium iodide (PI) staining and to quantify apoptotic cell death using PI staining and Annexin V/PI staining, respectively. The gene expression of p53, casp3, and Bcl-2 was measured using qRT-PCR. The Comet methodology was used to indicate DNA damage of yeast knockout strains.Results and conclusion The MTT assay revealed that CdCl2 was highly potent against hepatocellular carcinoma (HepG2), lung cancer (A549), and normal lung cell lines (Wi38) (IC50=3.12, 28.81, 191.14 μg/ml). These findings showed that CdCl2 reduced therapeutic efficacy in malignant cells at relatively low concentrations compared with nonmalignant cells, as well as confirmed the antitumor effect of the metal. CdCl2-damaged A549 cells revealed a significant increase in arrest of cell cycle in ‘S’ phases, and then apoptosis increased. Subsequently, when A549 cells were treated with a higher dosage of CdCl2, the gene expression of p53 and casp3 genes was upregulated, whereas Bcl-2 was downregulated. CdCl2 revealed its genotoxic activity at different concentrations of 10, 25, 50 μg/ml. MRE11, CLN1 and ZRC1 genes exhibited marked genotoxic effects, while MMT1 gene generated modret genotoxicity of yeast knockout strains. The Comet assay revealed that yeast cells were more responsive, which was indisputably demonstrated.

Nephroprotective effect of exercise training in cisplatin-induced renal damage in mice: influence of training protocol.

Cisplatin is an effective antineoplastic agent, but its use is limited by its nephrotoxicity caused by the oxidative stress in tubular epithelium of nephrons. On the other hand, regular exercise provides beneficial adaptations in different tissues and organs. As with many drugs, dosing is extremely important to get the beneficial effects of exercise. Thus, we aimed to investigate the influence of exercise intensity and frequency on cisplatin-induced (20 mg/kg) renal damage in mice. Forty male Swiss mice were divided into five experimental groups (n=8 per group): 1) sedentary; 2) low-intensity forced swimming, three times per week; 3) high-intensity forced swimming, three times per week; 4) low-intensity forced swimming, five times per week; and 5) high-intensity forced swimming, five times per week. Body composition, renal structure, functional indicators (plasma urea), lipid peroxidation, antioxidant enzyme activity, expression of genes related to antioxidant defense, and inflammatory and apoptotic pathways were evaluated. Comparisons considered exercise intensity and frequency. High lipid peroxidation was observed in the sedentary group compared with trained mice, regardless of exercise intensity and frequency. Groups that trained three times per week showed more benefits, as reduced tubular necrosis, plasma urea, expression of CASP3 and Rela (NFkB subunit-p65) genes, and increased total glutathione peroxidase activity. No significant difference in Nfe2l2 (Nrf2) gene expression was observed between groups. Eight weeks of regular exercise training promoted nephroprotection against cisplatin-mediated oxidative injury. Exercise frequency was critical for nephroprotection.

Tomentosin shows anticancer effect on U87 human glioblastoma multiforme cells

Glioblastoma multiforme (GBM) is one of the most common brain tumors. Chemotherapy, radiotherapy and surgical resection are methods used in GBM treatment, however, the investigation of possible anticancer effects of low-toxicity natural products on various cancer cells, including GBM, leads to promising results. In this study, it was aimed to investigate anticancer effect of tomentosin, which is a sesquiterpene lactone, on U87 human GBM cells. The cytotoxic effect of tomentosin was evaluated by XTT assay. The concentration of tomentosin that inhibits 50% cell viability (IC50) was determined by the results from XTT, and, in further analyzes cells were treated with tomentosin at IC50 concentration. Then, total RNA isolation and cDNA synthesis were performed in control and dose groups, and, the possible anticancer effect of tomentosin was determined by evaluating the expression levels of important genes associated with apoptosis and metastasis by qPCR analysis. In addition, the effect of tomentosin on the colony forming capacity of GBM cells was evaluated by colony formation assay. According to our results, IC50 dose of tomentosin was found to be 28.8 µM in U87 cells for 48 hours. When compared to the control group, tomentosin increased expression of BAX, CASP3, CASP8, CASP9, CYCS, FADD, TNF, TNFR1, TNFR2 and TIMP2 genes. And, tomantosin significantly decreased colony forming capacity of U87 cells. In conclusion, it is thought that tomentosin exerts its anticancer effect by changing the expression levels of genes associated with apoptosis and metastasis in U87 GBM cells.

Sorafenib, rapamycin, and venetoclax attenuate doxorubicin-induced senescence and promote apoptosis in HCT116 cells

Emerging evidence has shown that the therapy-induced senescent growth arrest in cancer cells is of durable nature whereby a subset of cells can reinstate proliferative capacity. Promising new drugs named senolytics selectively target senescent cells and commit them into apoptosis. Accordingly, senolytics have been proposed as adjuvant cancer treatment to cull senescent tumor cells, and thus, screening for agents that exhibit senolytic properties is highly warranted. Our study aimed to investigate three agents, sorafenib, rapamycin, and venetoclax for their senolytic potential in doxorubicin-induced senescence in HCT116 cells. HCT116 cells were treated with one of the three agents, sorafenib (5 µM), rapamycin (100 nM), or venetoclax (10 µM), in the absence or presence of doxorubicin (1 µM). Senescence was evaluated using microscopy-based and flow cytometry-based Senescence-associated-β-galactosidase staining (SA-β-gal), while apoptosis was assessed using annexin V-FITC/PI, and Muse caspase-3/-7 activity assays. We screened for potential genes through which the three drugs exerted senolytic-like action using the Human Cancer Pathway Finder PCR array. The three agents reduced doxorubicin-induced senescent cell subpopulations and significantly enhanced the apoptotic effect of doxorubicin compared with those treated only with doxorubicin. The senescence genes IGFBP5 and BMI1 and the apoptosis genes CASP7 and CASP9 emerged as candidate genes through which the three drugs exhibited senolytic-like properties. These results suggest that the attenuation of doxorubicin-induced senescence might have shifted HCT116 cells to apoptosis by exposure to the tested pharmacological agents. Our work argues for the use of senolytics to reduce senescence-mediated resistance in tumor cells and to enhance chemotherapy efficacy.

Bioactivity of betulinic acid nanoemulsions on skin carcinogenesis in transgenic mice K14E6

Alternative therapies for cancer treatment have been developed using bioactive compounds such as betulinic acid (BA). The objective of this study was to investigate the bioactivity of BA in its free form and compare it with its nano-encapsulated form under a skin carcinogenesis protocol in a genetically modified murine model. K14E6 and FVB mice were divided into four groups to be treated with free BA and with betulinic acid nanoemulsion (BANE). Lecithin enriched with medium chain fatty acids (MCFAs) was employed as an emulsifier to prepare the nanoemulsions with a mean droplet size of 40 nm. Skin tumors were induced by exposure to DMBA and TPA directly to the transgenic mice. Tumor development was completely inhibited by BANE and by 70% with free BA. This was validated by histological sections and the gene expression of the Cdk4 and Casp8 genes.

Generation of human long-lived plasma cells by developmentally regulated epigenetic imprinting.

Antibody secreting cells (ASCs) circulate after vaccination and infection and migrate to the BM where a subset known as long-lived plasma cells (LLPCs) persists and secrete antibodies for a lifetime. The mechanisms by which circulating ASCs become LLPCs are not well elucidated. Here, we show that human blood ASCs have distinct morphology, transcriptomes, and epigenetics compared with BM LLPCs. Compared with blood ASCs, BM LLPCs have decreased nucleus/cytoplasm ratio but increased endoplasmic reticulum and numbers of mitochondria. LLPCs up-regulate pro-survival genes MCL1, BCL2, and BCL-XL while simultaneously down-regulating pro-apoptotic genes HRK1, CASP3, and CASP8 Consistent with reduced gene expression, the pro-apoptotic gene loci are less accessible in LLPCs. Of the pro-survival genes, only BCL2 is concordant in gene up-regulation and loci accessibility. Using a novel in vitro human BM mimetic, we show that blood ASCs undergo similar morphological and molecular changes that resemble ex vivo BM LLPCs. Overall, our study demonstrates that early-minted blood ASCs in the BM microniche must undergo morphological, transcriptional, and epigenetic changes to mature into apoptotic-resistant LLPCs.

TNF signaling pathway-mediated microglial activation in the PFC underlies acute paradoxical sleep deprivation-induced anxiety-like behaviors in mice

Acute sleep deprivation is a common condition in modern life and increases anxiety symptoms in healthy individuals. The neuroinflammatory response induced by microglial activation could be an important contributing factor, but its underlying molecular mechanisms are still unclear. In the present study, we first found that acute paradoxical sleep deprivation (PSD) induced by the modified multiple platform method (MMPM) for 6 h led to anxiety-like behavior in mice, as verified by the open field test, elevated plus maze test, light–dark box test, and marble burying test. In addition, bioinformatic analysis suggested an important relationship between acute sleep deprivation and brain inflammatory signaling pathways. Key genes enriched in the TNF signaling pathway were confirmed to be altered during acute PSD by qPCR and Western blot analyses, including the upregulation of the prostaglandin-endoperoxide synthase 2 (Ptgs2) and suppressor of cytokine signaling 3 protein (Socs3) genes and the downregulation of the cysteine-aspartic acid protease 3 (Casp3) gene. Furthermore, we found that microglial cells in the prefrontal cortex (PFC) were activated with significant branch structure changes and that the cell body area was increased in the PSD model. Finally, we found that minocycline, a tetracycline with anti-inflammatory properties, may ameliorate the anxiogenic effect and microglial activation. Our study reveals significant correlations of anxiety-like behavior, microglial activation, and inflammation during acute PSD.

Silencing of α-N-acetylgalactosaminidase in the gastric cancer cells amplified cell death and attenuated migration, while the multidrug resistance remained unchanged.

Although the elevated level of the α-N-acetylgalactosaminidase enzyme (encoded by the NAGA gene) is a well-recognized feature of cancer cells; little research works have been undertaken on the cancer malignancy mechanisms. The effects of NAGA gene downregulation on cancer cells' features such as drug resistance, impaired programmed cell death, and migration were analyzed in this study. The cells grew exponentially with a doubling time of 30 h in an optimal condition. Toxicity of daunorubicin chemotherapy drug on NAGA-transfected EPG85.257RDB cells was evaluated in comparison to control cells and no significant change was recorded. Quantitative transcript analyses and protein levels revealed that the MDR1 pump almost remained unchanged during the study. Moreover, the NAGA gene downregulation enhanced the late apoptosis rate in EPG85.257RDB cells at 24 h posttransfection. The investigated expression level of genes and proteins involved in the TNFR2 signaling pathway, related to cancer cell apoptosis, showed considerable alterations after NAGA silencing as well. MAP3K14 and CASP3 genes were downregulated while IL6, RELA, and TRAF2 experienced an upregulation. Also, NAGA silencing generally diminished the migration ability of EPG85.257RDB cells and the MMP1 gene (as a critical gene in metastasis) expression decreased significantly. The expression of the p-FAK protein, which is located in the downstream of the α2 β1 integrin signaling pathway, was reduced likewise. It could be concluded that despite drug resistance, NAGA silencing resulted in augmentative and regressive effects on cell death and migration.

Effect of 4-Allyl-1-hydroxy-2-methoxybenzene (eugenol) in the expression of genes involved in cellular cycle and apoptotic process in dental pulp fibroblasts

Objective This study sought to evaluate the effect of eugenol on the cell morphology and expression of genes involved in the apoptotic process in human dental pulp fibroblasts (hDPFs) from deciduous teeth. Materials and methods hDPFs were cultured with 4 concentrations of eugenol (0.06 nM, 0.6 nM, 6 nM, 12 nM) and compared with a control group. After a 72 h incubation period, the cytotoxic effect on cell morphology by optical microscopy and gene expression by RT-PCR were evaluated. Results At 0.06 nM and 0.6 nM eugenol concentrations, vacuolisation of the cytoplasm was observed with atypical granulation of the hDPFs, and, at 6 nM and 12 nM cytoplasmic extensions disappeared almost completely. Casp-3, Casp-9, and telomerase genes were not expressed at the concentrations evaluated nor in the control group. The relative expression responses of Bcl-2 and TGF-β genes were overexpressed at the 4 concentrations. MAKP’s 0.06 nM (p < .001), 0.6 nM (p < .05) and 12 nM (p < .05) and Cyclin 1 at 12 nM showed significant difference versus the control group (p < .05). Conclusion Eugenol is capable of causing morphological changes in hDPFs in a dose-dependent manner, higher concentrations may promote overexpression of apoptotic genes.