Cellular P53 Research Articles

Identification of Novel Tumor-Microenvironment-Regulating Factor That Facilitates Tumor Immune Infiltration in Colon Cancer

Colon cancer is one of the most common malignancies causing death worldwide. It is well known that the cells of the tumor microenvironment contribute to the progression and prognosis of colon cancer. However, the gene alterations and potential remodeling mechanisms in the tumor microenvironment of colon cancer remain largely unknown. In this study, immune scores from the ESTIMATE algorithm were used to discriminate between patients with high or low immune-cell infiltration. There were 42 immune differentially expressed genes (DEGs) of prognostic value identified in this study. Among them, KCNJ5 is a key factor in promoting M2 macrophage recruitment and tumor immune infiltration in colon cancer. These findings may provide novel insights for decoding the complicated interplay between cancer cells and the tumor microenvironment as well as for developing new avenues for therapeutic intervention in colon cancer.

Repression of P53 Function by SIRT5-Mediated Desuccinylation at Lysine 120

p53 is a classic tumor suppressor that functions in maintaining genome stability by inducing either cell arrest for damage repair or cell apoptosis to eliminate damaged cells in response to different types of stress. Posttranslational modifications (PTMs) of p53 are thought to be the most effective way for modulating of p53 activation. Here, we show that SIRT5 interacts with p53 and suppresses its transcriptional activity. Using mass spectrometric analysis, we identify a previously unknown PTM of p53, namely, succinylation of p53 at Lysine 120 (K120), which occurs after DNA damage. SIRT5 catalyzes desuccinylation of p53 at K120, resulting in the suppression of p53 activation. Moreover, using double knockout mice, we validate that the suppression of p53 target gene expression and cell apoptosis upon DNA damage is dependent on cellular p53. Our study identifies a novel PTM of p53 that regulates its activation as well as reveals a new target of SIRT5 acting as a desuccinylase.

Rapha Myr®, a Blend of Sulforaphane and Myrosinase, Exerts Antitumor and Anoikis-Sensitizing Effects on Human Astrocytoma Cells Modulating Sirtuins and DNA Methylation.

Brain and other nervous system cancers are the 10th leading cause of death worldwide. Genome instability, cell cycle deregulation, epigenetic mechanisms, cytoarchitecture disassembly, redox homeostasis as well as apoptosis are involved in carcinogenesis. A diet rich in fruits and vegetables is inversely related with the risk of developing cancer. Several studies report that cruciferous vegetables exhibited antiproliferative effects due to the multi-pharmacological functions of their secondary metabolites such as isothiocyanate sulforaphane deriving from the enzymatic hydrolysis of glucosinolates. We treated human astrocytoma 1321N1 cells for 24 h with different concentrations (0.5, 1.25 and 2.5% v/v) of sulforaphane plus active myrosinase (Rapha Myr®) aqueous extract (10 mg/mL). Cell viability, DNA fragmentation, PARP-1 and γH2AX expression were examined to evaluate genotoxic effects of the treatment. Cell cycle progression, p53 and p21 expression, apoptosis, cytoskeleton morphology and cell migration were also investigated. In addition, global DNA methylation, DNMT1 mRNA levels and nuclear/mitochondrial sirtuins were studied as epigenetic biomarkers. Rapha Myr® exhibited low antioxidant capability and exerted antiproliferative and genotoxic effects on 1321N1 cells by blocking the cell cycle, disarranging cytoskeleton structure and focal adhesions, decreasing the integrin α5 expression, renewing anoikis and modulating some important epigenetic pathways independently of the cellular p53 status. In addition, Rapha Myr® suppresses the expression of the oncogenic p53 mutant protein. These findings promote Rapha Myr® as a promising chemotherapeutic agent for integrated cancer therapy of human astrocytoma.

DNA methylation status of bovine blastocysts obtained from peripubertal oocyte donors.

In the dairy industry, the high selection pressure combined with the increased efficiency of assisted reproduction technologies (ART) are leading toward the use of younger females for reproduction purposes, with the aim to reduce the interval between generations. This situation could impair embryo quality, decreasing the success rate of the ART procedures and the values of resulting offspring. Young Holstein heifers (n = 10) were subjected to ovarian stimulation and oocyte collection at 8, 11, and 14 months of age. All the oocytes were fertilized in vitro with semen from one adult bull, generating three pools of embryos per animal. Each animal was its own control for the evaluation of the effects of age. The EmbryoGENE platform was used to compare the DNA methylation status of blastocysts obtained from oocytes collected at 8 versus 14 and 11 versus 14 months of age. Age-related contrast analysis identified 5,787 and 3,658 differentially methylated regions (DMRs) in blastocysts from heifers at 8 versus 14 and 11 versus 14 months of age, respectively. For both contrasts, the DMRs were distributed nonrandomly in the different DNA regions. The DNA from embryos from 8-month-old donors was more hypermethylated, while the DNA from embryos from 11-month-old donors displayed an intermediate phenotype. According to Ingenuity Pathway Analysis, the upstream regulator genes cellular tumor antigen p53, transforming growth factor β1, tumor necrosis factor, and hepatocyte nuclear factor 4α are particularly associated with methylation sensitive targets, which were more hypermethylated in embryos from younger donors.

Thiocolchicoside and Colchicine Induced Apoptosis in Breast Cancer (MCF-7) Cells Via Up-regulated Expression of p53 Tumor Suppressor Protein Gene: An In vitro and In silico Docking Approaches

This study describes the antiproliferative activity of thiocolchicoside and colchicine compounds in a dose-dependent manner against breast cancer cell line (MCF-7). MTT assay results show that IC50 (inhibitory concentration 50 %) values towards MCF-7 breast cancer cell line were 79.02 and 74.23 nmol for thiocolchicoside and colchicines compounds, respectively. Further, in vitro interaction between thiocolchicoside and colchicine compound ligands binding with the p53 tumor suppressor protein gene and its expression was also confirmed by western blot analysis. Results clearly show that the expression of p53 protein was more abundant in treated MCF-7 cancer cell line and up-regulated expression of the p53 gene was positively correlated with thiocolchicoside and cochicine concentrations used. The thiocolchicoside compound was found to be more effective against p53 protein accumulation levels when compared with colchicine and adriamycin. Molecular docking analysis was performed to reveal the interactions of receptor binding sites (molecular targets) with ligands. These compounds show efficient inhibitory action on the target p53 tumor suppressor protein expression which confirms their increased anti-proliferative activity against cancer cell growth. The present results could be used for the improvement of a novel formulation with enhanced inhibition of MCF-7 cancer cell growth. The docking scores were found to be high in thiocolchicoside compound against cellular tumor antigen p53 (-5.9 kcal/mol) with the strong interaction followed by colchicine (-5.3 kcal/mol) and the hydrogen bond counts, confirming the capability of these compounds for binding with receptor active site. Therefore, thiocolchicoside and colchicine compounds with effective p53 inhibitory activity strongly suggest that these compounds could be used as strong anticancer agents to treat MCF-7 breast cancer cell growth arrest effectively.

Towards reconstructing the dipteran demise of an ancient essential gene: E3 ubiquitin ligase Murine double minute.

Genome studies have uncovered many examples of essential gene loss, raising the question of how ancient genes transition from essentiality to dispensability. We explored this process for the deeply conserved E3 ubiquitin ligase Murinedouble minute (Mdm), which is lacking in Drosophila despite the conservation of its main regulatory target, the cellular stress response gene p53. Conducting gene expression and knockdown experiments in the red flour beetle Tribolium castaneum, we found evidence that Mdm has remained essential in insects where it is present. Using bioinformatics approaches, we confirm the absence of the Mdm gene family in Drosophila, mapping its loss to the stem lineage of schizophoran Diptera and Pipunculidae (big-headed flies), about 95-85 million years ago. Intriguingly, this gene loss event was preceded by the de novo origin of the gene Companion of reaper (Corp), a novel p53 regulatory factor that is characterized by functional similarities to vertebrate Mdm2 despite lacking E3 ubiquitin ligase protein domains. Speaking against a 1:1 compensatory gene gain/loss scenario, however, we found that hoverflies (Syrphidae) and pointed-wing flies (Lonchopteridae) possess both Mdm and Corp. This implies that the two p53 regulators have been coexisting for ~ 150 million years in select dipteran clades and for at least 50 million years in the lineage to Schizophora and Pipunculidae. Given these extensive time spans of Mdm/Corp coexistence, we speculate that the loss of Mdm in the lineage to Drosophila involved further acquisitions of compensatory gene activities besides the emergence of Corp. Combined with the previously noted reduction of an ancestral P53 contact domain in the Mdm homologs of crustaceans and insects, we conclude that the loss of the ancient Mdm gene family in flies was the outcome of incremental functional regression over long macroevolutionary time scales.

A Network Pharmacology Approach to Identify Potential Molecular Targets for Cannabidiol's Anti-Inflammatory Activity.

Introduction: Published preclinical and clinical studies support the anti-inflammatory activity of CBD, but the molecular targets (e.g., genes and proteins) that are involved in its mechanisms of action remain unclear. Herein, a network-based pharmacology analysis was performed to aid in the identification of potential molecular targets for CBD's anti-inflammatory activity. Materials and Methods: Target genes and proteins were obtained from several online databases, including Swiss target prediction, Online Mendelian Inheritance in Man, and the DrugBank database. A compound-target-disease network was constructed with Cytoscape tool, and a network of protein-protein interactions was established with the Search Tool for the Retrieval of Interacting Genes/Proteins database. Lead proteins identified from the compound-target-disease network were further studied for their interactions with CBD by computational docking. In addition, biological pathways involved in CBD's anti-inflammatory activity were identified with the Gene Ontology enrichment and the Kyoto Encyclopedia of Genes and Genomes analysis. Results: A panel of proteins, including cellular tumor antigen p53, NF-kappa-B essential modulator, tumor necrosis factor (TNF) receptor, transcription factor p65, NF-kappa-B p105, NF-kappa-B inhibitor alpha, inhibitor of nuclear factor kappa-B kinase subunit alpha, and epidermal growth factor receptor, were identified as lead targets involved in CBD's anti-inflammatory activity. This finding was further supported by molecular docking, which showed interactions between the lead proteins and CBD. In addition, several signaling pathways, including TNF, toll-like receptor, mitogen-activated protein kinases, nuclear factor kappa-light-chain-enhancer of activated B cells, and nucleotide-binding oligomerization domain-like receptors, were identified as key regulators in the mediation of CBD's anti-inflammatory activity. Conclusion: A network-based pharmacology analysis identified potential molecular targets and signaling pathways for CBD's anti-inflammatory activity. Findings from this study add to the growing body of data supporting the utilization of CBD as a promising anti-inflammatory natural product.

Inhibition of Eukaryotic Translation Initiation Factor 5A (eIF5A) Hypusination Suppress p53 Translation and Alters the Association of eIF5A to the Ribosomes.

The eukaryotic translation initiation factor 5A (eIF5A) is an essential protein for the viability of the cells whose proposed function is to prevent the stalling of the ribosomes during translation elongation. eIF5A activity requires a unique and functionally essential post-translational modification, the change of a lysine to hypusine. eIF5A is recognized as a promoter of cell proliferation, but it has also been suggested to induce apoptosis. To date, the precise molecular mechanism through which eIF5A affects these processes remains elusive. In the present study, we explored whether eIF5A is involved in controlling the stress-induced expression of the key cellular regulator p53. Our results show that treatment of HCT-116 colon cancer cells with the deoxyhypusine (DHS) inhibitor N1-guanyl-1,7-diamineheptane (GC7) caused both inhibition of eIF5A hypusination and a significant reduction of p53 expression in UV-treated cells, and that eIF5A controls p53 expression at the level of protein synthesis. Furthermore, we show that treatment with GC7 followed by UV-induced stress counteracts the pro-apoptotic process triggered by p53 up-regulation. More in general, the importance of eIF5A in the cellular stress response is illustrated by the finding that exposure to UV light promotes the binding of eIF5A to the ribosomes, whereas UV treatment complemented by the presence of GC7 inhibits such binding, allowing a decrease of de novo synthesis of p53 protein.

Mini Review: Opposing Pathologies in Cancer and Alzheimer's Disease: Does the PI3K/Akt Pathway Provide Clues?

This minireview is a brief overview examining the roles of insulin-like growth factors (IGFs) and the PI3K/Akt pathway in two apparently unconnected diseases: Alzheimer's dementia and cancer. For both, increased age is a major risk factor, and, in accord with the global rise in average life expectancy, their prevalence is also increasing. Cancer, however, involves excessive cell proliferation and metastasis, whereas Alzheimer's disease (AD) involves cell death and tissue destruction. The apparent “inverse” nature of these disease states is examined here, but also some important commonalities in terms of the PI3K/Akt pathway, glucose utilization and cell deregulation/death. The focus here is on four key molecules associated with this pathway; notably, the insulin receptor substrate 1 (IRS-1), cellular tumor antigen p53 (p53), peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (PIN1) and low-density lipoprotein receptor–related protein-1 (LRP1), all previously identified as potential therapeutic targets for both diseases. The insulin-resistant state, commonly reported in AD brain, results in neuronal glucose deprivation, due to a dampening down of the PI3K/Akt pathway, including overactivity of the mammalian target of rapamycin 1 (mTORC1) complex, hyperphosphorylation of p53 and neuronal death. This contrasts with cancer, where there is overstimulation of the PI3K/Akt pathway and the suppression of mTORC1 and p53, enabling abundant energy and unrestrained cell proliferation. Although these disease states appear to be diametrically opposed, the same key molecules are controlling pathology and, with differential targeting of therapeutics, may yet provide a beneficial outcome for both.

Parallel CRISPR-Cas9 screens clarify impacts of p53 on screen performance.

CRISPR-Cas9 genome engineering has revolutionised high-throughput functional genomic screens. However, recent work has raised concerns regarding the performance of CRISPR-Cas9 screens using TP53 wild-type human cells due to a p53-mediated DNA damage response (DDR) limiting the efficiency of generating viable edited cells. To directly assess the impact of cellular p53 status on CRISPR-Cas9 screen performance, we carried out parallel CRISPR-Cas9 screens in wild-type and TP53 knockout human retinal pigment epithelial cells using a focused dual guide RNA library targeting 852 DDR-associated genes. Our work demonstrates that although functional p53 status negatively affects identification of significantly depleted genes, optimal screen design can nevertheless enable robust screen performance. Through analysis of our own and published screen data, we highlight key factors for successful screens in both wild-type and p53-deficient cells.

Radiomic prediction models for the level of Ki-67 and p53 in glioma.

ObjectiveTo identify glioma radiomic features associated with proliferation-related Ki-67 antigen and cellular tumour antigen p53 levels, common immunohistochemical markers for differentiating benign from malignant tumours, and to generate radiomic prediction models.MethodsPatients with glioma, who were scanned before therapy using standard brain magnetic resonance imaging (MRI) protocols on T1 and T2 weighted imaging, were included. For each patient, regions-of-interest (ROI) were drawn based on tumour and peritumoral areas (5/10/15/20 mm), and features were identified using feature calculations, and used to create and assess logistic regression models for Ki-67 and p53 levels.ResultsA total of 92 patients were included. The best area under the curve (AUC) for the Ki-67 model was 0.773 for T2 weighted imaging in solid glioma (sensitivity, 0.818; specificity, 0.833), followed by a less reliable AUC of 0.773 (sensitivity, 0.727; specificity 0.667) in 20-mm peritumoral areas. The highest AUC for the p53 model was 0.709 (sensitivity, 1; specificity, 0.4) for T2 weighted imaging in 10-mm peritumoral areas.ConclusionUsing T2-weighted imaging, the prediction model for Ki-67 level in solid glioma tissue was better than the p53 model. The 20-mm and 10-mm peritumoral areas in the Ki-67 and p53 model, respectively, showed predictive effects, suggesting value in further research into areas without conventional MRI features.

An Integrated Analysis of mRNA and lncRNA Expression Profiles Indicates Their Potential Contribution to Brown Fat Dysfunction With Aging.

Brown adipose tissue (BAT) can convert fatty acids and glucose into heat, exhibiting the potential to combat obesity and diabetes. The mass and activity of BAT gradually diminishes with aging. As a newly found regulator of gene expression, long non-coding RNAs (lncRNAs) exhibit a wide range of functions in life processes. However, whether long non-coding RNA (lncRNA) involves in BAT dysfunction with aging is still unclear. Here, using RNA-sequencing technology, we identified 3237 messenger RNAs (mRNAs) and 1312 lncRNAs as differentially expressed in BAT of 10-months-old mice compared with 6- to 8-week-old. The protein-protein interaction network and k-score analysis revealed that the core mRNAs were associated with two important aging-related pathways, including cell cycle and p53 signaling pathway. Gene set enrichment analysis indicated that these mRNAs might participate in lipid metabolism and brown fat dysfunction. Functional enrichment analyses demonstrated that dysregulated lncRNAs were associated with mitochondria, regulation of cellular senescence, cell cycle, metabolic and p53 signaling pathways. Moreover, we revealed that two lncRNAs (NONMMUT024512 and n281160) may involve in the regulation of their adjacent gene peroxisome proliferator-activated receptor alpha (Pparα), a thermogenesis regulator. Collectively, these results lay a foundation for extensive studies on the role of lncRNAs in age-related thermogenic degradation.

Mid and hindgut transcriptome profiling analysis of Atlantic salmon (Salmon salar) under unpredictable chronic stress.

The intestinal epithelium is a selectively permeable barrier for nutrients, electrolytes and water, while maintaining effective protection against pathogens. Combinations of stressors throughout an animal's life, especially in agriculture and aquaculture settings, may affect the regular operativity of this organ with negative consequences for animal welfare. In the current study, we report the effects of a three-week unpredictable chronic stress (UCS) period on the intestinal morphology and transcriptome response of Atlantic salmon (Salmon salar) parr midgut and hindgut. Midgut and hindgut from both control and UCS fish were collected for histology and RNA-sequencing analysis to identify respective changes in the membrane structures and putative genes and pathways responding to UCS. Histological analysis did not show any significant effect on morphometric parameters. In the midgut, 1030 genes were differentially expressed following UCS, resulting in 279 genes which were involved in 13 metabolic pathways, including tissue repair pathways. In the hindgut, following UCS, 591 differentially expressed genes were detected with 426 downregulated and 165 upregulated. A total of 53 genes were related to three pathways. Downregulated genes include cellular senescence pathways, p53 signalling and cytokine–cytokine receptor pathways. The overall results corroborate that salmon parr were at least partly habituating to the UCS treatment. In midgut, the main upregulation was related to cell growth and repair, while in the hindgut there were indications of the activated apoptotic pathway, reduced cell repair and inhibited immune/anti-inflammatory capacity. This may be the trade-off between habituating to UCS and health resilience. This study suggests possible integrated genetic regulatory mechanisms that are tuned when farmed Atlantic salmon parr attempt to cope with UCS.

KLF5 regulates epithelial-mesenchymal transition of liver cancer cells in the context of p53 loss through miR-192 targeting of ZEB2

ABSTRACT Krüppel-like factor 5 (KLF5) can both promote and suppress cell migration, but the underlying mechanisms have not been elucidated. In this study, we show that the function of KLF5 in epithelial-mesenchymal transition (EMT) and migration of liver cancer cells depends on the status of the cellular tumor antigen p53 (p53). Furthermore, zinc finger E-box-binding homeobox 2 (ZEB2) is the main regulator of KLF5 in EMT in liver cancer cells in the context of p53 loss. Most importantly, the regulation of ZEB2 by p53 and KLF5 is indirect and that miR-192 mediates this regulation. Finally, we find that in invasive liver cancer, KLF5 is absent in the context of p53 loss or mutation.

Protein-based virtual screening: SIRT-1 as modulator maturation of circulating endothelial progenitor cells

Circulating endothelial progenitor cells (cEPCs) are having involvement in the replacing vascular endothelial cells damaged or detachment from the basement membrane. cEPC needs to fulfill both quantity and quality requirements in order to play this important role. After efflux from bone marrow to circulation, niche EPC necessary of maturation to reach the potential for re-endothelialization. Risk factors exposure for cardiovascular disease not only affect the quantity and quality of cEPC, but it is also to be involved in downregulation of SIRT1. SIRT1 downregulation is mostly associated with the mechanism of senescence. Objective. to predict the direction of SIRT-1 interaction with cEPC maturation that is validated with cEPC marker surface using the STRING DB method of protein interaction. Result. SIRT1 interacts with two proteins, inhibiting VEGFA and activating and inhibiting P53. P53 inhibits the expression of p16ink4a, a protein involved in cellular senessence and P53 through AKT involved in CD 309 and ITGA2B expression. Whereas there was a direct interaction of the expression of CD 309 through the VEGFA line; CD117; TIE-2; CD 144; CD62a. Conclusion. SIRT-1 is an intracellular protein that is involved in cellular anti-aging processes but is not directly involved in the expression of EPC marker surfaces.

Clinicopathological study of potential biomarkers of Plasmodium falciparum malaria severity and complications

BackgroundEarly diagnosis, risk stratification and evaluation of possible biomarkers are much in demand nowadays, as the routine laboratory markers do not always predict severity of disease. The prevention of malaria caused by Plasmodium falciparum which when in severe form may lead to life threatening complications like acute kidney failure, heart disease, and respiratory altered etc. may be facilitated by these biomarkers. The present study was designed to evaluate the potential diagnostic biomarkers of P. falciparum malaria in complications. Materials and methodsSamples of P. falciparum (n = 74) and healthy control (n = 22) were collected from Dhalai district hospital, Dhalai, Tripura, India during September 2016 to October 2017 to study the potential biomarkers of the P. falciparum malaria severity and complications. 19 potential biomarkers of malaria complications and cellular signaling proteins were assessed using Enzyme Linked Immuno Sorbent assays (ELISA) and biochemical analyses for selected proteins. Mann -Whitney U test and chi square test were used to compare the malaria case versus healthy control group and uncomplicated malaria versus severe malaria group. The prognostic performance for all parameters was assessed using ROC (receiver operating characteristic) analysis. ResultsThe demographic, clinical and laboratory parameters were significantly altered in case group compared to healthy controls, p-value <.05 except gender, cholesterol, triglycerides, Gamma-GT and Cytochrome c. The respiratory rate, CK-MB, uric acid, CKD-EPI eGFR, total bilirubin, Caveolin1 and pLDH were significantly altered in the severe malaria group as compared to the uncomplicated malaria group (p-value<.05). ConclusionInterestingly, the known markers of heart ailment CK-MB and PGC1α, kidney dysfunction marker CKD-EPI eGFR and other cellular signaling markers, DAPK1, p53 and Beclin1 were significantly altered in uncomplicated malaria than healthy controls. These findings may pave path for unfolding of disease pathogenesis and complexity in malaria infection.

Hormone-Like Effects of Bisphenol A on p53 and Estrogen Receptor Alpha in Breast Cancer Cells.

Bisphenol A (BPA) is a polymerizing agent commonly found in plastics that has been linked to xenoestrogenic activity. In this study, we analyzed the estrogen-like effects of BPA on the expression of estrogen receptor (ER)α and p53 with hormonal and antihormonal treatments in T-47D and MCF-7 cells. Cells were cultured in medium containing 5% charcoal-stripped fetal bovine serum for 6 days to deplete any endogenous steroids or effectors. The cells were then treated for 24 h with 600 nM BPA, which was determined to be the optimal value by a concentration study of BPA from 1 nM to 2 μM. Extracted cellular proteins were quantified and subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE)/Western blot analysis. The cell proliferation assays were quantified upon exposure to BPA. Laser confocal microscopy was performed to determine the cytolocalization of p53 and ERα upon treatment with BPA. Western blot analysis revealed that BPA caused an increase in the cellular protein p53 in a concentration-dependent manner. While treatment with BPA did not affect the cytolocalization of p53, an increase in cell proliferation was observed. Our studies provide interesting leads to delineate the possible mechanistic relationship among BPA, ER, and tumor suppressor proteins in breast cancer cells.

Network pharmacology—Deciphering the molecular mechanism of San-Zi-Yang-Qin decoction for the treatment of chronic obstructive pulmonary disease

IntroductionSan-Zi-Yang-Qin Decoction (SZYQD) is a common traditional Chinese herbal medicine, which has been used for centuries in the treatment of Chronic Obstructive Pulmonary Diseases (COPD). In order to investigate the molecular mechanism of SZYQD in the treatment of COPD, we analyzed the effective active ingredients, potential targets, cooperative functions and associated pathways based on network pharmacology. MethodsActive ingredients were collected and identified to predict their targets from TCMSP database. Predicted targets related to COPD were screened with CTD to establish a Drugs-components-targets network and Proteins interactions network using STRING Database and Cytoscape software. Action targets were highlighted by pictures of correlative pathways using KEGG PATHWAY Database. Results29 effectively active ingredients were filtered out, including Stigmasterol, beta-carotene, beta-sitosterol, luteolin, linolenic acid, METHYL LINOLEATE, etc. There were 170 targets predicted and all of them were related to COPD, including Heme oxygenase 1 (HMOX1), Glutathione S-transferase P 1 (GSTP1), Phosphatidylinositol 3-kinase (PI3K), Apoptosis regulator Bcl-2 (Bcl-2), Extracellular regulated protein kinases (ERK), Transcription factor p65 (RELA), NF-kappa-B inhibitor alpha (NFKBIA), Matrix metalloprotease (MMP), Cellular tumor antigen p53 (TP53), Caspase, Muscarinic acetylcholine receptor M1-5 (CHRM1-5), etc, enriched with a number of relevant GO functions and KEGG pathways. ConclusionSZYQD may regulate the Cholinergic synapse, Th17 cell differentiation, Fluid shear stress and atherosclerosis, Apoptosis, PI3K-Akt signaling pathway, NF-κB signaling pathway, TNF signaling pathway, C-type lectin receptor signaling pathway and so on, to resist oxidation, diminish inflammation, balance proliferation and apoptosis of cells, inhibit vagal excitation in the treatment of COPD.

Fucoidan from marine brown algae attenuates pancreatic cancer progression by regulating p53 – NFκB crosstalk

Poor pancreatic cancer (PC) prognosis has been attributed to its resistance to apoptosis and propensity for early systemic dissemination. Existing therapeutic strategies are often circumvented by the molecular crosstalk between cell-signalling pathways. p53 is mutated in more than 50% of PC and NFκB is constitutively activated in therapy-resistant residual disease; these mutations and activations account for the avoidance of cell death and metastasis. Recently, we demonstrated the anti-PC potential of fucoidan extract from marine brown alga, Turbinaria conoides (J. Agardh) Kützing (Sargassaceae). In this study, we aimed to characterize the active fractions of fucoidan extract to identify their select anti-PC efficacy, and to define the mechanism(s) involved. Five fractions of fucoidan isolated by ion exchange chromatography were tested for their potential in genetically diverse human PC cell lines. All fractions exerted significant dose-dependent and time-dependent regulation of cell survival. Fucoidans induced apoptosis, activated caspase −3, −8 and −9, and cleaved Poly ADP ribose polymerase (PARP). Pathway-specific transcriptional analysis recognized inhibition of 57 and 38 nuclear factor κB (NFκB) pathway molecules with fucoidan-F5 in MiaPaCa-2 and Panc-1 cells, respectively. In addition, fucoidan-F5 inhibited both the constitutive and Tumor necrosis factor-α (TNFα)-mediated NFκB DNA-binding activity in PC cells. Upregulation of cytoplasmic IκB levels and significant reduction of NFκB-dependent luciferase activity further substantiate the inhibitory potential of seaweed fucoidans on NFκB. Moreover, fucoidan(s) treatment increased cellular p53 in PC cells and reverted NFκB forced-expression-related p53 reduction. The results suggest that fucoidan regulates PC progression and that fucoidans may target p53–NFκB crosstalk and dictate apoptosis in PC cells.

Non-invasive detection of EGFR and TP53 mutations through the combination of plasma, urine and sputum in advanced non-small cell lung cancer.

The sensitivity and utility of liquid biopsy in clinical practice requires some improvement. The aim of the present study was to improve the detection of epidermal growth factor (EGFR) and cellular tumor antigen p53 (TP53) mutations in liquid biopsies from patients with advanced non-small cell lung cancer (NSCLC) by combining plasma, sputum and urine samples under the same sequencing platform. Plasma, sputum and urine samples, and tumor tissues were obtained from 50 patients with NSCLC and were analyzed using next-generation sequencing. The sensitivity of EGFR-sensitive mutation detection was 84% in plasma, 63% in sputum, 28% in urine, and 91% when combining the three liquid samples (P<0.001). The sensitivity of TP53 mutation detection increased from 87% in plasma to 94% when the three samples were combined (P<0.001). The sensitivity of EGFR or TP53 mutations detection was higher in patients with multiple metastatic sites compared with patients ≤1 metastatic site. In addition, the progression free survival (PFS) rates obtained following analysis of the three samples independently in patients with EGFR sensitizing mutations were similar, and were 9.0 months in the tissue sample, 7.5 months in plasma, 7.9 months in the sputum and 7.3 months in urine (P=0.721). The PFS of patients with TP53 mutations was shorter compared with patients without TP53 mutations and was as follows: Tissue, 8.2 months compared with 10.2 months (P=0.412); plasma, 8.4 months compared with 10.2 months (P=0.466); sputum, 8.3 months compared with 9.1 months (P=0.904); and when combined, 8.8 months compared with 10.3 months (P=0.599). The combination of plasma, sputum and urine increased the detection of EGFR or TP53 mutation with higher sensitivity, and may improve the predictive value of personalized treatment.