Gene P53 Research Articles

Enzymatic and transcriptional level changes induced by the co-presence of lead and procymidone in hook snout carp (Opsariichthys bidens)

Understanding the interactions between different environmental pollutants is necessary in ecotoxicology since environmental contaminants never appear as single components but rather in combination with other substances. Heavy metals and pesticides are commonly detected in the environment, but the characterization of their mixture toxicity has been inadequately explored. This research aimed to elucidate the mixture impacts of the heavy metal lead (Pb) and the pesticide procymidone (PCM) on the hook snout carp (Opsariichthys bidens) using an array of biomarkers. The data showed that Pb and PCM possessed almost equivalent acute toxicity to the animals, with 4-days LC50 values of 120.9 and 85.15 mg L−1, respectively. Combinations of Pb and PCM generated acute synergistic effects on O. bidens. The contents of malondialdehyde (MDA), antioxidative (SOD), apoptotic (caspase-9), and detoxifying enzymes glutathione S-transferase (GST) and cytochrome P450 (CYP450) significantly changed after most of the mixture exposures compared with the baseline level and the corresponding individual exposures. This suggests the induction of oxidative stress, cell damage, and detoxification dysfunction. The expressions of eight genes (mn-sod, cu-sod, p53, cas3, erβ1, esr, ap, and klf2α) associated with oxidative stress, cell apoptosis, immune response, and hormonal functions exhibited pronounced changes when challenged with the mixture compared to the individual treatments. This indicates the occurrence of immune dysregulation and endocrine disorder. These findings provide an overall understanding of fish upon the challenge of sublethal toxicity between Pb and PCM and can be adopted to evaluate the complicated toxic mechanisms in aquatic vertebrates when exposed to heavy metal and pesticide mixtures. Additionally, these results might guide environmental regulation tactics to protect the population of aquatic vertebrates in natural ecosystems.

Abstract B079: Identification of synergistic drug combinations including MEK inhibitor and agents modulating the tumor microenvironment in an electroporation-induced pancreatic cancer model

Abstract Pancreatic ductal adenocarcinoma (PDAC) is refractory to cytostatic and immuno-oncology therapeutic strategies, leaving surgical resection of the tumor as the only treatment that can significantly prolong survival. We are therefore exploring optimized treatment strategies targeting cancer cells as well as infiltrating immune cells and the tumor microenvironment (TME) in an autochthonous mouse model for PDAC. Single focal tumors are induced through in vivo electroporation of gene constructs into the pancreas which encode mutated Kras and facilitate knock out the tumor suppressor genes P53, Cdkn2a and/or Smad4 via CRISPR/Cas9. The resulting tumors recapitulate the human disease with respect to genetics, histology and TME composition. Furthermore, this model allows the generation of tumors reflecting different PDAC subtypes, as well as the resection of primary tumors in combination with the (neo-)adjuvant regimens. Like the human disease, these tumors are refractory to chemotherapy and immune checkpoint blockade, whereas targeted therapy by means of MEK inhibitors merely leads to temporary delay of tumor outgrowth. We therefore tested the combination of MEK inhibitor treatment with different, potentially complementary modalities. We found the combination with agonist anti-CD40 antibodies to be highly effective. Prominent regressions are observed, and in combination with primary tumor resection long-term complete responses can be achieved even in a metastatic variant of the model. Apart from the direct cytostatic impact of MEKi on the Kras-transformed tumor cells, the mechanism of action of this synergistic drug combination involves enhancement of the anti-tumor T-cell response, suppression of CD4+ Tregs, and reduction of MDSCs and M2-type macrophages in favor of M1-type pro-inflammatory macrophages. A second highly effective regimen as identified in our experiments involves the combination of MEK inhibitors with the multi-kinase inhibitor regorafenib. In contrast to the aforementioned regimen, the complementary action of these drugs involves modulation of the TME by regorafenib. While the clinical translation of the MEKi/CD40 Ab regimen is hampered by toxicity issues encountered with agonist anti-CD40 Ab, the synergy as observed in the MEKi/regorafenib regiment provides a tangible path towards clinical testing. Citation Format: Aline S. Konrad, Tanja Hägele, Stefan Zens, Caroline Vent, Moritz Hamberger, Sarah Baiker, Aron Maes, Carl-Stephan Leonhardt, Oliver Strobel, Oliver Politz, Dieter Zopf, Daniel Baumann, Julian Mochayedi, Rienk Offringa. Identification of synergistic drug combinations including MEK inhibitor and agents modulating the tumor microenvironment in an electroporation-induced pancreatic cancer model [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B079.

MiR-155-5p promotes hepatic steatosis via PICALM-mediated autophagy in aging hepatocytes

BackgroundHepatic steatosis, a lipid disorder characterized by the accumulation of intrahepatic fat, is more prevalent in the elderly population. This study investigates the role of miR-155–5p in the autophagy dysregulation of aging hepatic steatosis. MethodsWe established an aging mouse model in vivo and a hepatocellular senescence model induced by low serum and palmitic acid in vitro. The fluctuations of microRNAs were derived from RNA-seq data and confirmed by qPCR in 4- and 18-month-old mouse liver tissues. Hematoxylin-eosin (H&E) staining observed pathological changes. Markers of senescence, autophagy, and lipolysis genes were analyzed using Western blot and qPCR. Bioinformatics analysis predicted miR-155–5p's target gene PICALM, confirmed by dual luciferase reporter assay and transfection of miR-155–5p mimic/inhibitor into senescent hepatocytes. ResultsSenescent markers (p21, p16, and p-P53) and miR-155–5p were up-regulated in aging liver tissues and senescent hepatocytes. Bioinformatics analysis identified PICALM as a target gene of miR-155–5p, a finding further supported by dual luciferase reporter assays. Inhibition of miR-155–5p reduced expression of senescent marker genes (p16, p21, p-P53), improved autophagy (evidenced by increased LC3B-II and ATG5, and decreased P62), and enhanced lipolysis (indicated by increased ATGL and p-HSL) in senescent hepatocytes. Oil red O staining confirmed that miR-155–5p inhibition significantly reduced lipid accumulation in these cells. ConclusionsThis study suggests a potential new therapeutic approach for age-related hepatic steatosis through the inhibition of miR-155–5p to enhance autophagy.

İsatinin HepG2 ve AML12 hücre hatları üzerindeki moleküler etkilerinin araştırılması

Isatin is an indole-derived organic compound. It is a natural component of the Couroupita guianensis plant. It is also the metabolic derivative of the human body hormone adrenaline. Studies have shown the anti-tumoral effects of isatin derivatives. In this study, the cytotoxic effects of isatin on HepG2, a hepatocellular cancer cell line was investigated. Additionally, its cytotoxic and protective-proliferative effects on AML12, a healthy liver cell line was investigated. This evaluation was conducted using MTT, fluorescent staining, wound healing, and real-time polymerase chain reaction analyses. The IC50 values for 48 hours of isatin application were calculated as 186.23 µM for HepG2 and 7.05 mM for AML12. The analysis of wound healing and fluorescent staining at varying doses of HepG2 application revealed suppression of proliferation and triggered apoptosis in HepG2 cells. In contrast, AML12 cells exhibited promoted proliferation under similar conditions. Moreover, the observed upregulation of oxidative stress genes CuZn/Mn-SOD and mitochondrial apoptotic pathway genes Bax, cleaveled-Cas3, APAF1, and p53 in HepG2 cells contrasted with their decreased expression in AML12 cell lines. These results suggest the potential of natural isatin as a promising anti-cancer agent for liver cancer cell lines and as a protective supplement for healthy liver cells.

Cadmium-induced annulus fibrosus cell senescence contributes to intervertebral disc degeneration via the JNK/p53 signaling pathway.

Investigating the impact of cadmium (Cd) on annulus fibrosus (AF) cells and its potential mechanism was the purpose of the current study. Cd was cultivated in different concentrations (0, 1, 5, 10, and 20 μM) on AF cells and the potential effects of the metal were assessed. Using the CCK-8 method, cell viability and proliferation were identified. Using transcriptome analysis, the annulus fibrosus cells were sequenced both with and without cadmium chloride. The EdU method was used to determine the rate of cell proliferation; senescence-associated β-galactosidase (SA-β-Gal) staining was used to determine the number of positive cells; and western blot, RT-PCR, and immunofluorescence were used to determine the protein and mRNA expression of senescence-associated proteins (p16, p21, and p53) and c-Jun N-terminal kinase (JNK). According to the findings, Cd has the ability to increase the production of senescence-associated genes (p16 and p21) and senescence-associated secreted phenotype (SASP), which includes IL-1β and IL-6. Through the JNK/p53 signal pathway, Cd exposure simultaneously accelerated AF cell senescence and promoted SASP. Following JNK inhibitor (SP600125) treatment, the expression of p53, JNK, and senescence-associated indices were all down-regulated. By activating the JNK/p53 signaling pathway, Cd can induce oxidative stress damage and AF cell senescence. These findings could provide a new approach for treating and preventing intervertebral disc degeneration (IVDD) caused by Cd exposure.

Proliferation and Differentiation Potential of Bone Marrow-Derived Mesenchymal Stem Cells From Children With Polydactyly and Adults With Basal Joint Arthritis.

This study compared the proliferation and differentiation potential of bone marrow-derived mesenchymal stem cells (BMSCs) derived from infants with polydactyly and adults with basal joint arthritis. The proliferation rate of adult and infant BMSCs was determined by the cell number changes and doubling times. The γH2AX immunofluorescence staining, age-related gene expression, senescence-associated β-galactosidase (SA-β-gal) staining were analyzed to determine the senescence state of adult and infant BMSCs. The expression levels of superoxide dismutases (SODs) and genes associated with various types of differentiation were measured using Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR). Differentiation levels were evaluated through histochemical and immunohistochemical staining. The results showed that infant BMSCs had a significantly higher increase in cell numbers and faster doubling times compared with adult BMSCs. Infant BMSCs at late stages exhibited reduced γH2AX expression and SA-β-gal staining, indicating lower levels of senescence. The expression levels of senescence-related genes (p16, p21, and p53) in infant BMSCs were also lower than in adult BMSCs. In addition, infant BMSCs demonstrated higher antioxidative ability with elevated expression of SOD1, SOD2, and SOD3 compared with adult BMSCs. In terms of differentiation potential, infant BMSCs outperformed adult BMSCs in chondrogenesis, as indicated by higher expression levels of chondrogenic genes (SOX9, COL2, and COL10) and positive immunohistochemical staining. Moreover, differentiated cells derived from infant BMSCs exhibited significantly higher expression levels of osteogenic, tenogenic, hepatogenic, and neurogenic genes compared with those derived from adult BMSCs. Histochemical and immunofluorescence staining confirmed these findings. However, adult BMSCs showed lower adipogenic differentiation potential compared with infant BMSCs. Overall, infant BMSCs demonstrated superior characteristics, including higher proliferation rates, enhanced antioxidative activity, and greater differentiation potential into various lineages. They also exhibited reduced cellular senescence. These findings, within the context of cellular differentiation, suggest potential implications for the use of allogeneic BMSC transplantation, emphasizing the need for further in vivo investigation.

Polyethylene microplastics affect behavioural, oxidative stress, and molecular responses in the Drosophila model.

The escalating presence of microplastic pollution poses a significant environmental threat, with far-reaching implications for both ecosystems and human health. This study investigated the toxicological impact of polyethylene microplastics (PE MPs) using Drosophila melanogaster, fruit flies, as a model organism. Drosophila were exposed to PE MPs orally at concentrations of 1 mg ml-1 and 10 mg ml-1 agar food. The study assessed behavioural parameters and biochemical markers including reactive oxygen species (ROS), superoxide dismutase (SOD), and glutathione-S-transferase (GST) activity. The expression levels of key genes (Hsp70Bc, rpr, and p53) were also analysed using the RT-qPCR technique. Results indicated a significant decline in climbing activity among adult flies and crawling behaviour in larvae, indicating potential disruption of motor function. Biochemical analysis revealed elevated ROS levels, indicative of oxidative stress, in both larval and fly stages. Moreover, the antioxidant defence system exhibited decreased SOD activity and a concentration-dependent increase in GST activity indicating the functioning of a quick xenobiotic clearance mechanism. Gene expression analysis demonstrated upregulation of rpr, p53, and Hsp70Bc genes, suggesting activation of cell death pathways and stress response mechanisms. Overall, these findings underline the adverse effects of PE MPs on Drosophila, including behavioural impairment, oxidative stress, and activation of stress response pathways.

Discovery of new 1,3-diphenylurea appended aryl pyridine derivatives as apoptosis inducers through c-MET and VEGFR-2 inhibition: design, synthesis, in vivo and in silico studies.

Interest has been generated in VEGFR-2 and c-MET as potential receptors for the treatment of different malignancies. Using aryl pyridine derivatives with 1,3-diphenylurea attached, a number of promising dual VEGFR-2 and c-MET inhibitors were developed and synthesized. Regarding the molecular target, compounds 2d, 2f, 2j, 2k, and 2n had potent IC50 values of 65, 24, 150, 170, and 18 nM against c-MET, respectively. Additionally, they had potent IC50 values of 310, 35, 290, 320, and 24 nM against VEGFR-2, respectively. Regarding cytotoxicity, compounds 2d, 2f, 2j, 2k and 2n exhibited potent cytotoxicity against MCF-7 with IC50 values in the range 0.76-21.5 μM, and they showed promising cytotoxic activity against PC-3 with IC50 values in the range 1.85-3.42 μM compared to cabozantinib (IC50 = 1.06 μM against MCF-7 and 2.01 μM against PC-3). Regarding cell death, compound 2n caused cell death in MCF-7 cells by 87.34-fold; it induced total apoptosis by 33.19% (8.04% for late apoptosis, 25.15% for early apoptosis), stopping their growth in the G2/M phase, affecting the expression of apoptosis-related genes P53, Bax, caspases 3 and 9 and the anti-apoptotic gene, Bcl-2. In vivo study illustrated the anticancer activity of compound 2n by reduction of tumor mass and volume, and the tumor inhibition ratio reached 56.1% with an improvement of hematological parameters. Accordingly, compound 2n can be further developed as a selective target-oriented chemotherapeutic against breast cancer.

Investigation on the signaling pathways in the mechanism of hyperoxia-induced acute lung injury based on transcriptomics sequencing

To observe and verify the changes of transcriptome in hyperoxia-induced acute lung injury (HALI), and to further clarify the changes of pathways in HALI. Twelve healthy male C57BL/6J mice were randomly divided into normoxia group and HALI group according to the random number table, with 6 mice in each group. The mice in the normoxia group were fed normally in the room, and the mice in the HALI group was exposed to 95% oxygen to reproduce the HALI animal model. After 72 hours of hyperoxia exposure, the lung tissues were taken for transcriptome sequencing, and then Kyoto Encyclopedia of Genes and Genomes database (KEGG) pathway enrichment analysis was performed. The pathological changes of lung tissue were observed under light microscope after hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to verify the key molecules in the signal pathways closely related to HALI identified by transcriptomics analysis. Transcriptomic analysis showed that hyperoxia induced 537 differentially expressed genes in lung tissue of mice as compared with the normoxia group including 239 up-regulated genes and 298 down-regulated genes. Further KEGG pathway enrichment analysis identified 20 most significantly enriched pathway entries, and the top three pathways were ferroptosis signaling pathway, p53 signaling pathway and glutathione (GSH) metabolism signaling pathway. The related genes in the ferroptosis signaling pathway included the up-regulated gene heme oxygenase-1 (HO-1) and the down-regulated gene solute carrier family 7 member 11 (SLC7A11). The related genes in the p53 signaling pathway included the up-regulated gene tumor suppressor gene p53 and the down-regulated gene murine double minute 2 (MDM2). The related gene in the GSH metabolic signaling pathway was up-regulated gene glutaredoxin 1 (Grx1). The light microscope showed that the pulmonary alveolar structure of the normoxia group was normal. In the HALI group, the pulmonary alveolar septum widened and thickened, and the alveolar cavity shrank or disappeared. RT-RCR and Western blotting confirmed that compared with the normoxia group, the mRNA and protein expressions of HO-1 and p53 in lung tissue of the HALI group were significantly increased [HO-1 mRNA (2-ΔΔCt): 2.16±0.17 vs. 1.00±0.00, HO-1 protein (HO-1/β-actin): 1.05±0.01 vs. 0.79±0.01, p53 mRNA (2-ΔΔCt): 2.52±0.13 vs. 1.00±0.00, p53 protein (p53/β-actin): 1.12±0.02 vs. 0.58±0.03, all P < 0.05], and the mRNA and protein expressions of Grx1, MDM2, SLC7A11 were significantly decreased [Grx1 mRNA (2-ΔΔCt): 0.53±0.05 vs. 1.00±0.00, Grx1 protein (Grx1/β-actin): 0.54±0.03 vs. 0.93±0.01, MDM2 mRNA (2-ΔΔCt): 0.48±0.03 vs. 1.00±0.00, MDM2 protein (MDM2/β-actin): 0.57±0.02 vs. 1.05±0.01, SLC7A11 mRNA (2-ΔΔCt): 0.50±0.06 vs. 1.00±0.00, SLC7A11 protein (SLC7A11/β-actin): 0.72±0.03 vs. 0.98±0.01, all P < 0.05]. HALI is closely related to ferroptosis, p53 and GSH metabolism signaling pathways. Targeting the key targets in ferroptosis, p53 and GSH metabolism signaling pathways may be an important strategy for the prevention and treatment of HALI.

Supplementation of Saussurea costus root alleviates sodium nitrite-induced hepatorenal toxicity by modulating metabolic profile, inflammation, and apoptosis.

Sodium nitrite (NaNO2) is a widely used food ingredient, although excessive concentrations can pose potential health risks. In the present study, we evaluated the deterioration effects of NaNO2 additives on hematology, metabolic profile, liver function, and kidney function of male Wistar rats. We further explored the therapeutic potential of supplementation with S. costus root ethanolic extract (SCREE) to improve NaNO2-induced hepatorenal toxicity. In this regard, 65 adult male rats were divided into eight groups; Group 1: control, Groups 2, 3, and 4 received SCREE in 200, 400, and 600mg/kg body weight, respectively, Group 5: NaNO2 (6.5mg/kg body weight), Groups 6, 7 and 8 received NaNO2 (6.5mg/kg body weight) in combination with SCREE (200, 400, and 600mg/kg body weight), respectively. Our results revealed that the NaNO2-treated group shows a significant change in deterioration in body and organ weights, hematological parameters, lipid profile, and hepatorenal dysfunction, as well as immunohistochemical and histopathological alterations. Furthermore, the NaNO2-treated group demonstrated a considerable increase in the expression of TNF-α cytokine and tumor suppressor gene P53 in the kidney and liver, while a significant reduction was detected in the anti-inflammatory cytokine IL-4 and the apoptosis suppressor gene BCL-2, compared to the control group. Interestingly, SCREE administration demonstrated the ability to significantly alleviate the toxic effects of NaNO2 and improve liver function in a dose-dependent manner, including hematological parameters, lipid profile, and modulation of histopathological architecture. Additionally, SCREE exhibited the ability to modulate the expression levels of inflammatory cytokines and apoptotic genes in the liver and kidney. The phytochemical analysis revealed a wide set of primary metabolites in SCREE, including phenolics, flavonoids, vitamins, alkaloids, saponins and tannins, while the untargeted UPLC/T-TOF-MS/MS analysis identified 183 metabolites in both positive and negative ionization modes. Together, our findings establish the potential of SCREE in mitigating the toxic effects of NaNO2 by modulating metabolic, inflammatory, and apoptosis. Together, this study underscores the promise of SCREE as a potential natural food detoxifying additive to counteract the harmful impacts of sodium nitrite.

The prognostic impact of EpCAM and p53 expressions in infiltrating duct carcinoma of the breast and their association with the clinicopathological prognostic parameters

Background Breast cancer is the most common cause of cancer death among women worldwide. Adhesion molecule EpCAM is an important player in carcinogenesis; however, its exact biological role is not clear. Tumor suppressor gene p53 maintains genomic stability. In malignant cells, its function can be compromised by various mechanisms such as mutations, alteration of p53 regulators, and alteration of p53 target genes. This study aimed to find the relationship between epithelial cell adhesion molecule (EpCAM) and p53 expressions on one hand and clinicopathological factors of prognostic importance in breast cancer on the other hand and find any correlations between EpCAM and p53 markers. Materials and methods Immunohistochemical staining with EpCAM and p53 was studied on 42 cases of infiltrating duct carcinoma. Results A significant association was found between EpCAM and tumor size (P=0.049), tumor grade (P=0.005), lymph node status (P=0.043), pathological tumor stage; pathological tumor stage (P=0.027), and lymphovascular invasion (P=0.048). However, EpCAM was negatively associated with estrogen receptor (P=0.007), progesterone receptor (P=0.031), and human epidermal growth factor receptor 2/neu (P=0.025) expressions. Expression of p53 was positively associated with tumor size (P=0.025), tumor grade (P=0.002), lymph node status (P=0.05), pathological tumor stage (P=0.041), and lymphovascular invasion (P=0.043), while negatively associated with estrogen receptor (P=0.029), progesterone receptor (P=0.043), and human epidermal growth factor receptor 2/neu (P=0.039) expressions. A positive association was found between EpCAM and p53 expressions and triple-negative breast cancer (P=0.007 and 0.029, respectively). There was a positive association between EpCAM and p53 (P=0.007). Conclusion EpCAM and p53 expressions are good markers of predicting poorer outcomes in breast cancer patients. There is a positive association suggesting a combined prognostic value in breast cancer and it may confirm the binding of p53 with the EpCAM gene to regulate its function.

Analyzing the Expression of MicroRNA-375 and its Target Gene p53 in Oral Squamous Cell Carcinoma and its Implication in Oral Carcinogenesis

Objective: Oral mucosal cancers are the 11th most common human malignancies worldwide with a five-year survival rate of ≤50%. The lacunae of reliable diagnostic and prognostic markers pose an enormous challenge to the timely identification and prediction of disease progression in oral cancer. MicroRNAs (miRNAs) are emerging molecular markers associated with cancer initiation, progression, and therapy. The present study evaluated the microRNA -375(miR-375) expression and its target p53 gene in Oral squamous cell carcinoma (OSCC) to validate its utility as a diagnostic marker of the disease. Patients and Methods: This case-control study targeted histopathologically diagnosed cases of OSCC. miR-375 was quantified from 22 cases of OSCC and corresponding control tissues using qRT-PCR. Mutant p53 expression in cases and controls was determined by subjecting the tissues to immunohistochemical Results: Significant downregulation of miR-375 was noted in OSCC tissues (68.1%) compared to the control tissues with a mean fold change of 83.9 (p&lt;0.05). Significant downregulation of miR-375 was noted in Paan and tobacco chewing patients (77.8%). Men exhibited considerable downregulation compared to women (p&lt;0.05). The miR-375 expression levels did not correlate with the patient’s age, location, size, nodal status, and histopathological grade of the tumor. About 63.6 % of OSCC tissues showed mutant p53 positivity. Mutant p53 expression was noted in 73.3% of miR-375 downregulated tumors. Smokers exhibited higher expression of mutant p53 contrary to non-smokers(p&lt;0.00). P53 immunopositivity showed a correlation with tumor size, histopathological grade, and nodal metastasis. Conclusion: The findings of the study indicate that miR-375 downregulation may have a crucial effect on oral carcinogenesis by targeting p53. miR-375 should be further evaluated as a potential marker for oral cancer diagnosis.

Association and Functional Analysis of Ferroptosis-related Genes with Prostate Hyperplasia

Objective: This study aimed to investigate the association and biological functions of ferroptosis-related genes with prostatic hyperplasia. Methods: Gene information from ferroptosis and prostatic hyperplasia databases was used to screen for ferroptosis-related genes associated with prostate hyperplasia. Protein-protein interaction (PPI), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and enrichment analysis were employed to evaluate the interaction between these genes and prostatic hyperplasia. Results: Through PPI, GO, KEGG, and enrichment analysis, several ferroptosis-related genes associated with prostatic hyperplasia were identified, including phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), mitogen-activated protein kinase 3 (MAPK3), GABA type A receptor associated protein like 1 (GABARAPL1), HRas proto-oncogene (HRAS), tumor suppressor gene p53 (TP53), MAPK1, and autophagy related protein 7 (ATG7). These genes were found to be involved in various biological processes and signaling pathways related to ubiquitin protein ligase binding, protein phosphatase binding, pre-autophagosomal structure, autophagy, the PI3K-Akt signaling pathway and so on. Core genes such as PIK3CA, MAPK3, GABARAPL1, HRAS, TP53, MAPK1, ATG7, EGFR, Kirsten rat sarcoma viral oncogene homolog, and coiled-coil myosin-like BCL2-interacting protein were also found to be associated with transcription factors MYC, ESR2, PML, JUN, and TP53. Protein drug prediction suggested that FTI-277, Ginkgolide A, Tipifarnib, Desvenlafaxine, and Defactinib could potentially target the core genes. Conclusion: This study revealed the association and biological functions of ferroptosis-related genes with prostatic hyperplasia, providing new clues and directions for understanding the pathophysiology of prostatic hyperplasia and selecting therapeutic targets. These findings also contributed to a deeper understanding of the relationship between iron metabolism and prostatic hyperplasia, providing new ideas and methods for the diagnosis and treatment of iron metabolism-related diseases.

The potential use of ascorbic acid to recover the cellular senescence of lipopolysaccharide-induced human apical papilla cells: an in vitro study.

To examine the effect of lipopolysaccharide (LPS) on cellular senescence induction of human apical papilla cells (hAPCs) and evaluate the potential use of 50 μg/ml ascorbic acid to recover cellular senescence and regenerative functions. hAPCs were treated with LPS at 1 and 10 μg/ml either with or without 50 μg/ml ascorbic acid for 48 h. The cellular senescence biomarkers were analyzed by senescence-associated β-galactosidase (SA-β-gal) staining and senescence-related gene expression, p16 and p21. Cell migration, at 12 h and 24 h, was evaluated using a scratch wound assay. Mineralization potential was assessed at 21 days using Alizarin red S staining and dentine sialophosphoprotein (DSPP) and bone sialoprotein (BSP) gene expression. 1 μg/ml and 10 μg/ml LPS stimulation for 48 h induced cellular senescence, as shown by remarkable SA-β-gal staining and p16 and p21 gene expression. The percentage of wound closure and mineralized formation was reduced. The co-incubation with ascorbic acid significantly down-regulated the level of SA-β-gal staining. The reduction of senescence-associated gene expressions was observed. Ascorbic acid improved cell migration, mineralized nodule formation, and the expression of DSPP and BSP genes in LPS-treated hAPCs. LPS significantly promoted cellular senescence on hAPCs and diminished the cell function capacity. Co-presence of ascorbic acid could impede cellular senescence and possibly improve the regenerative capacity of LPS-induced senescent hAPCs in vitro. The data support the in vitro potential benefit of ascorbic acid on cellular senescence recovery of apical papilla cells.

Combined absence of TRP53 target genes ZMAT3, PUMA and p21 cause a high incidence of cancer in mice

Transcriptional activation of target genes is essential for TP53-mediated tumour suppression, though the roles of the diverse TP53-activated target genes in tumour suppression remains poorly understood. Knockdown of ZMAT3, an RNA-binding zinc-finger protein involved in regulating alternative splicing, in haematopoietic cells by shRNA caused leukaemia only with the concomitant absence of the PUMA and p21, the critical effectors of TRP53-mediated apoptosis and cell cycle arrest respectively. We were interested to further investigate the role of ZMAT3 in tumour suppression beyond the haematopoietic system. Therefore, we generated Zmat3 knockout and compound gene knockout mice, lacking Zmat3 and p21, Zmat3 and Puma or all three genes. Puma−/−p21−/−Zmat3−/− triple knockout mice developed tumours at a significantly higher frequency compared to wild-type, Puma−/−Zmat3−/− or p21−/−Zmat3−/−deficient mice. Interestingly, we observed that the triple knockout and Puma−/−Zmat3−/− double deficient animals succumbed to lymphoma, while p21−/−Zmat3−/− animals developed mainly solid cancers. This analysis suggests that in addition to ZMAT3 loss, additional TRP53-regulated processes must be disabled simultaneously for TRP53-mediated tumour suppression to fail. Our findings reveal that the absence of different TRP53 regulated tumour suppressive processes changes the tumour spectrum, indicating that different TRP53 tumour suppressive pathways are more critical in different tissues.

Biogenic Zinc oxide nanoparticles from Celosia argentea: toward improved antioxidant, antibacterial, and anticancer activities.

Biogenic Zinc oxide (ZnO) nanoparticles (NPs) were synthesized from Celosia argentea (C. argentea) plant extract. Structural analysis confirms the successful synthesis of biogenic zinc oxide NPs from C. argentea extract. The biogenic ZnO NPs have an average particle size of 21.55 ± 4.73nm, a semispherical shape, and a specific surface area of about 50m2/g. The biogenic ZnO NPs have a powerful radical scavenging activity (Ic50 = 91.24mg/ml) comparable to ascorbic acid (ASC) as a standard (Ic50 = 14.37mg/ml). The antibacterial efficacy was tested against gram-positive and gram-negative bacteria using an agar disc diffusion method. Gram-positive strains with biogenic ZnO NPs have a greater bactericidal impact than gram-negative strains in a concentration-dependent manner. Anticancer activity against Liver hepatocellular cells (HepG2) and Human umbilical vein endothelial cells (HUVEC) was evaluated using a [3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide] (MTT) assay. The results reflect the concentration-dependent cytotoxic effect of biogenic ZnO NPs against HepG2 cells even at low concentrations (Ic50 = 49.45μg/ml) compared with doxorubicin (Ic50 = 14.67μg/ml) and C. argentea extract (Ic50 = 112.24μg/ml). The cell cycle and gene expression were analyzed to determine the potential anticancer mechanism. The flow cytometric analysis of the cell cycle revealed that biogenic ZnO NPs induce oxidative stress that activates the apoptotic genes NF-κB, CY-C, and P53, leading to cell death. The Celosia argentea improved the antioxidant, antibacterial, and anticancer activities of ZnO NPs without altering their structural properties. The effect of green synthesis on the bioactivity of biogenic ZnO NPs in vivo is recommended for future work.

Nanofibers based on zein protein loaded with tungsten oxide for cancer therapy: fabrication, characterization and in vitro evaluation

Plant proteins have become attractive for biomedical applications such as wound dressing and drug delivery. In this research, nanofibers from pristine zein (plant protein) and zein loaded with tungsten oxide (WO3) were prepared (WO3@zein) using less toxic solvents (ethanol and acetic acid). Morphological and biological properties of the zein nanofiber were determined. Prepared nanofibers were defined by thermogravimetric analysis (TGA), X-ray diffraction (X-RD), Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy. The average fiber diameter was unchanged with an increase in WO3 concentration from 0.001 to 0.008%. FT-IR spectroscopy and X-RD indicated the presence of WO3 in WO3@zein nanofibers. In comparison to WO3-free, WO3@zein nanofibers showed higher safety and preserved the anticancer effect of WO3 against human melanoma cell line (A375) melanoma cells compared to WO3-free. Moreover, both WO3-free and WO3@zein caused a fourfold increase in the cellular proliferation of reactive oxygen species (ROS) in the treated A375 cells compared to untreated cells. ROS elevation led to apoptosis-dependent cell death of A375 cells as evidenced by up-regulating the expression of p53-downstream genes (p21 and Bax) (tumor-suppressor gene) while down-regulating the expression of key oncogenes (BCL2 and cyclin D). In conclusion, the prepared nanofiber represents a promising and safe candidate for anticancer applications.

Investigating The effect of Particulate Matter (PM2.5) On The expression Profile and Methylation of EGR Gene Family and P53 Gene in The lung Tissue of Rat Exposed to Air Pollution

Investigating The effect of Particulate Matter (PM2.5) On The expression Profile and Methylation of EGR Gene Family and P53 Gene in The lung Tissue of Rat Exposed to Air Pollution

Doxorubicin induced epigenetic regulation of dendritic cell maturation in association with T cell activation facilitates tumor protective immune response in non-small cell lung cancer (NSCLC)

BackgroundNSCLC is one of the leading causes of death and is often diagnosed at late stages with no alternative therapeutic approach. DCs are professional antigen-presenting cells and DC-based immunotherapy has been under the spotlight for its anti-cancer properties. Epigenetic modifications including DNA methylation and histone modification in DCs play a crucial role in regulating their functions such as maturation and activation,innate immune responses, T cell priming, antigen presentation, and cytokine production. In the current study, we investigated the anti-cancer properties of Doxorubicin at a noncytotoxic concentration that could be extrapolated as an epigenetic regulator for DC maturation to elicit anti-tumor activity. MethodologiesPBMCs from normal and NSCLC blood samples were isolated and treated with growth factors. DCs were matured with low dose Doxorubicin and the DC maturation markers were checked by using flow-cytometry. Further, ELISA was performed and low dose Doxorubicin-induced DCs were pulsed with LCA (Lung Cancer Antigen) and primed with CD4 +T helper (Th) cells for cytotoxicity assessment. Further, epigenetic markers of T: DC conjugation were immunofluorescently visualized under a microscope. ChIP-qPCR and Invitro assays such as histone methylation, DNA methylation, and m6A methylation were performed to study the epigenetic changes under low dose Dox treatment. IL-12 neutralization assay was performed to check for the IL-12 dependency of DCs and their effect under Dox at low dose treatment. This was further followed by a Western Blotting analysis for histone and non-histone proteins. ResultsLow dose Doxorubicin induces epigenetic changes in DCs to elicit an anti-tumor response in NSCLC through the generation of CTLs with a concomitant increase in the extracellular secretions of anti-inflammatory cytokines. We also found that low dosage of Doxorubicin matured DCs when pulsed with LCA and primed with CD4 +T helper cells, secrete IFN-γ which is important in orchestrating adaptive immunity by activating CD8 + cytotoxic T-lymphocytes. Also, the secretions of IL-12 help us infer that protective immunity is also induced via Th1 response which triggered selectively the translocation of PKCθ to immunological synapse in between DC and Th. Further, methylation and acetylation markers H3K4me3 and H3K14Ac respectively upregulated whereas levels of STAT5, NFkB, NOTCH1, and DNAPKcs were downregulated. DNA and RNA methylation assays then lead to confirmations about the epigenetic changes caused by low dose Dox treatment. DNA methylation was reduced which resulted in the activation of tumor suppressor gene p53 and Th1-associated transcription factor TBX21. On the other hand, both absolute and relative RNA methylation quantification increased in the presence of Dox at a low dose. ConclusionFrom this study, we understand that non-cytotoxic concentration of Doxorubicin increases the Ag-presenting ability of DCs via an IL-12-dependent mechanism and causes epigenetic modifications in NSCLC.

Combined effects of hypoxia and ammonia-N exposure ontheimmune response, oxidative stress, tissue injury and apoptosis of hybrid grouper (Epinephelus fuscoguttatus♀×E. lanceolatus♂).

In order to investigate the simultaneous exposure to hypoxia and ammonia-N on oxidative stress, immune response, and apoptosis of the hybrid grouper, 120 healthy groupers were selected for hypoxia and/or ammonia-N exposure experiment. The fish were divided into four experimental groups: hypoxia and ammonia-N group, hypoxia group, ammonia-N group, and control group. The results demonstrated that ammonia-N and hypoxia exposures induced the superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities increased first and then decreased, and malondialdehyde (MDA) accumulated. Additionally, antioxidant genes (SOD, CAT, GSH-Px, HSP70, and HSP90), apoptosis genes (p53, bax, caspase 3, caspase 8, and caspase 9), and inflammatory genes (TNF-α, IL-1β, IL-6, and IL-8) were upregulated by hypoxia and ammonia-N exposure. Severe inflammatory features were noticed in fish under hypoxia and ammonia-N co-exposure and speculating that the p53-bax pathway may induce apoptosis in hybrid groupers. Furthermore, hybrid grouper exposed to hypoxia or ammonia-N revealed some abnormalities in liver histology, with combined exposure resulting in the most severe liver tissue lesions. In summary, the hypoxia and ammonia-N co-exposure induced oxidative stress, accelerating the cell damage and activated inflammation and apoptosis.