Upregulation Of Reactive Oxygen Species Research Articles

Transcriptomics profile reveals the temporal molecular events triggered by cut-wounding in stem-ends of cut ‘Tiber’ lily flowers

Plants are prone to wounding by various means, which mainly occur with exposure to environmental mechanical stimuli or via herbivory. The plant wounding response is believed to be mediated by potential molecular mechanisms. However, the wounding response and its molecular mechanism in cut flowers, which are severely wounded plant tissues, have been poorly studied. To gain insight into the molecular mechanisms in response to cut-wounding, this work presents the transcriptional characterization of the temporal molecular events triggered by cut-wounding in the stem-ends of cut ‘Tiber’ lily flowers. RNA-Seq analysis detected 11887 differentially expressed genes, which could be categorized into four clusters based on response time and expression patterns. The fast response typically includes the up-regulation of reactive oxygen species (ROS) and jasmonic acid (JA) biosynthesis genes within 1–6 h after cut-wounding, followed by the up-regulation of lignin biosynthesis genes. Further analysis revealed complex signal cascades in lily stems following cut-wounding. In the early stage (1–6 h), Ca2+ promotes ROS and JA biosynthesis gene expression through calcium signal transmission. In the late stage (6–48 h), the expression of lignin biosynthesis genes was up-regulated by ROS and JA signal transduction. These results provide insight into the temporal molecular events in lily stems triggered by cut-wounding and their potential molecular mechanisms.

Protective effects of mannan/β-glucans from yeast cell wall on the deoxyniyalenol-induced oxidative stress and autophagy in IPEC-J2 cells

The aim of this study was to investigate the effects of biomacromolecules mannan/β-glucans from yeast cell wall (BYCW) to alleviate Deoxynivalenol(DON)-induced injury. Considering that DON has strong oxidizing effect and stimulates autophagy and apoptosis, we examined the effects of BYCW on consequent oxidative stress damage indicators, cells autophagy and apoptosis induced by DON using the porcine jejunum epithelial cell lines (IPEC-J2) as a cell culture model. The results showed that application of BYCW could reverse the decrease of cell viability by DON significantly, and suppress the levels of tumor necrosis factor-α (TNF-α) and interleukin-8 and -6 (IL-8 and IL-6), except IL-1β. Further experiments revealed that BYCW treatment counteracted the DON-induced down-regulation of intracellular glutathione (GSH) and up-regulation of reactive oxygen species (ROS) and malondialdehyde (MDA). Through western blot analysis, we observed that BYCW treatment was able to down-regulate the expression of autophagy protein LC3-II and up-regulate the expression of P62 protein against DON, which suggested that autophagy induced by DON may be suppressed. Altogether, these results indicated a potential ability of supplementation of BYCW to improve cell growth and metabolism as well as the preventive properties of BYCW against the DON-induced cell damage by activating antioxidant system.

<p>SIRT6 promotes the Warburg effect of papillary thyroid cancer cell BCPAP through reactive oxygen species</p>

Purpose: Our previous study demonstrated that SIRT6 is upregulated in papillary thyroid cancer (PTC) and enhances tumor aggressiveness. In this study, we further researched its influence in the Warburg effect.Methods: SIRT6-upregulated and downregulated BCPAP cells and negative control BCPAP-NC groups were generated with lentiviral vectors. In these two cell lines, reactive oxygen species (ROS) were detected by dichlorodihydrofluorescein diacetate. Expression of the key Warburg effect genes including GLUT1, HK2, GAPDH, PGK1, ENO1, PKM2 and LDHA was measured by quantitative real-time PCR and western blotting. Glucose uptake, lactate production and the ATP content of cells were detected with assay kits. The ROS scavenger N-acetylcysteine was used for treatment of BCPAP-SIRT6, and the same measurements as described above were detected again.Results: Compared with the BCPAP-NC group, expression of the key Warburg effect genes including Glut1, HK2 and GAPDH and their protein products was upregulated in the BCPAP-SIRT6 group, whereas BCPAP-shSIRT6 showed significant downregulation. Meanwhile, ROS, glucose uptake, lactate production and ATP content of the BCPAP-SIRT6 group were also significantly increased, and BCPAP-shSIRT6 showed significant downregulation. Furthermore, upregulation of key Warburg effect genes and glucose uptake, lactate production and ATP content were all rescued after treatment with ROS scavenger.Conclusion: SIRT6 promoted the Warburg effect of PTC cells via upregulation of ROS. Inhibition of ROS in SIRT6-upregulated cells could rescue activation of the Warburg effect.

Osteogenic and chondrogenic differentiation effect of neohesperidin dihydrochalcones in human adipose‐derived stem cells

Reactive oxygen species (ROS) are highly reactive which cause damage to various cellular functions such as proliferation, differentiation, and survival. Nuclear factor erythroid 2‐related factor‐2 (Nrf2) promotes expression of antioxidant enzyme such as heme oxygenase 1 (HO‐1) and NAD(P)H:Quinone Oxidoreductase 1 (NQO‐1). Neohesperidin is a compound originated from peel of citrus fruits and its hydrogenated form, neohesperidin dihydrochalcone (NHDC) has antioxidant activity. Human adipose‐derived stem cells (hADSC) are multipotent adult stem cells responsible for the regeneration of fat, bone and cartilage. Thus, we observed the antioxidant effects of NHDC derivatives on hADSC through Nrf2 activation. Upregulation of ROS inhibits osteogenic differentiation of hADSCs, in part through inhibition of signaling pathway, which is essential for bone development and maintenance. ROS levels regulate inhibition of proliferation and modulate initiation of the hypertrophic changes in chondrocytes. Cellular viability and compounds' proper concentrations were set by MTS assay. The effects of these compounds on suppressing oxidative stress and hADSCs differentiation into osteocytes and chondrocytes was investigated using Alizarin Red staining, Alcian Blue staining, reverse‐transcriptase polymerase chain reaction (RT‐PCR), and Western blotting. The results demonstrate that NHDC derivatives play an efficacious role in inducing osteogenic and chondrogenic differentiation of hADSCs through reducing cellular oxidative stress.Support or Funding InformationThis research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education 2018R1D1A1B07049077This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

CRP-Induced NLRP3 Inflammasome Activation Increases LDL Transcytosis Across Endothelial Cells.

The NLRP3 inflammasome, a multiprotein cytosolic complex that activates the IL-1 family of cytokines, plays an important role in atherosclerosis (AS). High-sensitivity c-reactive protein (hs-CRP) is widely recognized as a major cardiovascular risk predictor and recent studies name NLRP3 as a predictor of CRP levels. Mounting evidence has indicated that subendothelial retention of apolipoprotein B100-containing lipoproteins, such as low-density lipoprotein (LDL), is the initial step of atherogenesis, and is usually termed the “response to retention hypothesis.” We previously reported that CRP promotes AS by directly increasing LDL transcytosis across endothelial cells (ECs). The present study aims to investigate the effects of CRP on NLRP3 inflammasome activation and the role of the NLRP3 inflammasome in CRP-induced LDL transcytosis. We found that CRP upregulated NF-κB activity, the NF-κB inhibitor (BAY-11-7082) and Fcγ receptors (FcγRs) inhibitor (CD32/64Ab) blocked CRP-induced NF-κB activation. CRP also induced expression of pro-IL-1β and NLRP3, while BAY and CD32/64 Ab suppressed CRP-mediated expression of NLRP3 and pro-IL-1β. Moreover, CRP activated the NLRP3 inflammasome in ECs. NADPH oxidase inhibitor, diphenylene iodonium (DPI) and dithiothreitol (DTT), a broad-spectrum P2 receptor inhibitor, oxidized ATP (oATP), and a broad inhibitor of cysteine proteases, E-64d, inhibited CRP-induced NLRP3 inflammasome activation. Furthermore, NLRP3 siRNA and caspase-1 inhibitor blocked CRP-mediated LDL transcytosis across ECs. In conclusion, NLRP3 inflammasome activation was shown to be involved in CRP-mediated LDL transcytosis across ECs. CRP not only increased the expression of pro-IL-1β and NLRP3 via the FcγRs/NF-κB pathway, but also promoted NLRP3 inflammasome activation and IL-1β maturation by upregulation of reactive oxygen species (ROS) levels, purinergic receptor signaling, and activation of cysteine proteases.

2,3-butanediol Induces Systemic Acquired Resistance in the Plant Immune Response

Plants harbor a wide diversity of microorganisms, which are involved in major plant functions such as nutrition and resistance to biotic and abiotic stresses. Recently, the importance of the rhizosphere microbiome for plant growth has been widely recognized. Therefore, we researched the effects of 2,3-butanediol (2,3-BD) in order to obtain insights into systemic acquired resistance (SAR) mediated through reactive oxygen species (ROS) homeostasis and pathogenesis-related (PR) gene expression. Syringe infiltration with Paenibacillus polymyxa DSM 365 surprisingly mitigated cell damage, which was induced by the compatible plant pathogen Phytophtora parasitica var. nicotianae (Ppn). Furthermore, syringe infiltration with 2,3-BD produced from P. polymyxa effectively enhanced SAR to compatible Ppn through down-regulation of ROS biosynthetic genes (NtRbohD and NtRbohF) and up-regulation of ROS detoxification and PR protein expression. In addition, synergy between 2,3-BD and nonexpressor pathogenesis-related protein 1 (NPR1) enhanced resistance to pathogen infection. Taken together, our study demonstrates the potential applicability of leaf and root-associated microbiomes as biopestcides to increase efficiency and yield in agricultural systems.

Arginine-Rich Manganese Silicate Nanobubbles as a Ferroptosis-Inducing Agent for Tumor-Targeted Theranostics.

Ferroptosis, an iron-based cell-death pathway, has recently attracted great attention owing to its effectiveness in killing cancer cells. Previous investigations focused on the development of iron-based nanomaterials to induce ferroptosis in cancer cells by the up-regulation of reactive oxygen species (ROS) generated by the well-known Fenton reaction. Herein, we report a ferroptosis-inducing agent based on arginine-rich manganese silicate nanobubbles (AMSNs) that possess highly efficient glutathione (GSH) depletion ability and thereby induce ferroptosis by the inactivation of glutathione-dependent peroxidases 4 (GPX4). The AMSNs were synthesized via a one-pot reaction with arginine (Arg) as the surface ligand for tumor homing. Subsequently, a significant tumor suppression effect can be achieved by GSH depletion-induced ferroptosis. Moreover, the degradation of AMSNs during the GSH depletion contributed to T1-weighted magnetic resonance imaging (MRI) enhancement as well as on-demand chemotherapeutic drug release for synergistic cancer therapy. We anticipate that the GSH-depletion-induced ferroptosis strategy by using manganese-based nanomaterials would provide insights in designing nanomedicines for tumor-targeted theranostics.

Trimetazidine Protects Against Atherosclerosis by Changing Energy Charge and Oxidative Stress.

BackgroundThis study investigated the effect and the possible mechanism of trimetazidine in atherosclerosis.Material/MethodsWe established an atherosclerotic rat model by high-fat diet and vitamin D injection. Rats were separated into 3 different groups: control, atherosclerosis, and trimetazidine (n=10). The aortic artery was isolated and its morphological features were examined by hematoxylin and eosin (HE) staining. Serum low-density lipoprotein cholesterol (LDL-c), total cholesterol (TC), and triglycerides (TG) were analyzed using an automatic biochemical analyzer. Human aortic smooth muscle cells (HASMCs) were cultured and divided into 5 groups: no treatment, H2O2 treatment only, trimetazidine preincubation before H2O2 treatment, oxidized low-density lipoprotein (oxLDL) treatment only, and trimetazidine preincubation before oxLDL treatment. HASMCs proliferation was tested using the Cell Counting Kit-8. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity of the aortic artery, and HASMCs were measured using commercially available kits.ResultsHE staining assay showed that trimetazidine suppressed the progression of atherosclerosis and reduced foam cell formation in the aortic artery without affecting serum lipid levels. HASMCs proliferation assay revealed that trimetazidine alleviated the inhibitory effect of H2O2 on HASMCs proliferation and inhibited oxLDL-induced proliferation of HASMCs. Moreover, trimetazidine ameliorated ROS up-regulation elicited by H2O2 or oxLDL in HASMCs. Additionally, trimetazidine restored SOD activity and reduced MDA content of HASMCs.ConclusionsTrimetazidine suppressed the progression of atherosclerosis by enhancing energy value, decreasing ROS level of aortic artery, modulating HASMCs proliferation, and reducing oxidative stress in HASMCs.

Co-delivery of siAlox15 and sunitinib for reversing the new-onset of type 1 diabetes in non-obese diabetic mice

Type 1 diabetes (T1D) is a chronic autoimmune disease caused by proinflammatory T cell infiltration associated cytokine secretion and reactive oxygen species (ROS) production. In this study, our objective was to determine whether co-delivery of sunitinib, a member of tyrosine kinase inhibitor (TKI) family, and siRNA against Alox15 (siAlox15) could reverse the new-onset of T1D in non-obese diabetic (NOD)/ShiLtJ female mice which spontaneously develop diabetes. A cationic copolymer methoxy poly(ethylene glycol)-block-poly(2-methyl-2-carboxyl-propylene carbonate-graft-dodecanol-graft-tetraethylenepentamine) (mPEG-b-PCC-g-DC-g-TEPA) was used to encapsulate sunitinib with 8.32-9.39% loading efficacy and form complex with siAlox15 at the N/P ratio of 16:1. These micellar formulations exhibited sustained release of sunitinib for around 50% at 24 h and protected siAlox15 from serum degradation for at least 10 h. Further, both sunitinib and siAlox15 inhibited human peripheral blood mononuclear cell (PBMC) proliferation and activation by downregulating the phosphorylation of Akt, platelet-derived growth factor receptor (PDGFR) at protein levels and Alox15 in mRNA levels, respectively. Besides, siAlox15 treatment reversed the cytokine induced ROS upregulation in rat INS-1E β cells. Finally, systemic administration of these micelles carrying sunitinib and siAlox15 efficiently reversed T1D in NOD mice as indicated by decreased blood glucose concentration to below 250 mg/dL. The mice showed fast response to high glucose which was determined by intraperitoneal glucose tolerance test and promoted serum insulin production. Tissue staining indicated that sunitinib and siAlox15 combination could inhibit T cell infiltration to mouse islets, thereby promoting islet survival and function. In summary, our results demonstrated that combination therapy of sunitinib and siAlox15 could be a novel, reliable and safe approach for patients who were recently diagnosed with T1D.

Plumbagin inhibits the proliferation of nasopharyngeal carcinoma 6-10B cells by upregulation of reactive oxygen species.

Plumbagin (PLB) is the primary component of the traditional Chinese medicine Baihua Dan, and possesses anti-infection and anticancer effects, with the ability to enhance the sensitivity of tumor cells to radiation therapy. However, the anticancer effect of PLB on nasopharyngeal carcinoma and the underlying mechanisms remain unclear. In this study, we investigated the anticancer effects of PLB on nasopharyngeal carcinoma 6-10B cells and clarified its molecular mechanisms in vitro. The results showed that PLB was effective against 6-10B cells proliferation in a dose-dependent manner by inducing G2/M phase cell cycle arrest. Furthermore, our data showed that PLB induced reactive oxygen species accumulation, which inhibited the GSK3β/STAT3 pathway and arrested the G2/M phase. Therefore, our results provided new insight into the mechanism of the antitumor effects of PLB, supporting PLB as a prospective therapeutic drug in nasopharyngeal carcinoma by modulating intracellular redox balance.

Protective effects ROS up-regulation on premature ovarian failure by suppressing ROS-TERT signal pathway.

Premature ovarian failure (POF) refers to the condition of pre-onset ovarian function failure, and is one commonly occurred disease in gynecology. Its pathogenic mechanism, however, is still unclear. Early study found decreased activity of telomerase reverse transcriptase (TERT). As an important factor to suppress TERT, oxidative stress has not been studied in POF. We, thus, investigated the role of reactive oxygen species (ROS)-TERT in POF. Rat POF model was induced by a single intraperitoneal injection of cyclophosphamide plus 12 mg/kg busulfan. Level of follicle stimulating hormone (FSH) and inhibin B was measured by enzyme-linked immunosorbent assay (ELISA), along with hematoxylin and eosin (HE) staining to confirm successful generation of models. Western blot was applied to measure TERT expression, and N-acetyl-cysteine (NAC) or TERT small interfere RNA (siRNA) was injected to suppress ROS or TERT level, followed by HE staining to observe POF condition. In POF model, ovary tissues showed atrophy, less follicles, and more follicular atresia, plus mesenchymal hyperplasia. FSH and inhibin B level were significantly up-regulated and down-regulated, respectively (p<0.05). In POF rat, ROS level was elevated (p<0.05) whilst TERT level was decreased. NAC inhibited ROS level and enhanced TERT expression. In contrast, TERT siRNA further aggravated POF condition. ROS up-regulation inhibits TERT expression, suppresses TERT activity and facilitates POF. The ROS-TERT pathway may work as the target for treating POF.

SAHA (vorinostat) facilitates functional polymer-based gene transfection via upregulation of ROS and synergizes with TRAIL gene delivery for cancer therapy

Non-viral gene delivery is an attractive approach for the treatment of many diseases including cancer, benefiting from its safety and large-scale production concerns. However, the relatively low transfection efficacy compared with viral vectors restricts the clinical applications of non-viral gene vectors. Reactive oxygen species (ROS) triggered charge reversal polymers (named B-PDEAEA) presented improved transfection efficacy, because of fast release of plasmid DNA responding to enhanced oxidative stress in cancer cells. But inadequate dissociation can still occur owing to the insufficient intracellular ROS generation. Here, we report SAHA (vorinostat), which is a clinical histone deacetylase inhibitor and anticancer drug, induces the ROS accumulation in cancer cells, and facilitates the charge reversal process of B-PDEAEA and the cellular dissociation of the delivered gene from the vectors. As a result, SAHA remarkably increases the gene transfection efficacy in an ROS-dependent manner. Importantly, SAHA synergizes with B-PDEAEA mediated therapeutic gene TNF-related apoptosis-inducing ligand (TRAIL) delivery in inducing apoptosis of cancer cells. These findings support the first concept of improving the gene delivery efficacy of stimuli-responsive vectors through upregulating the cellular ROS via an FDA approved anticancer agent. Additionally, combination of SAHA and TRAIL gene therapy could be a potential strategy for cancer treatment.

Deletion of REDD1 Prevents Hyperglycemia-Induced Reactive Oxygen Species Accumulation and Retinal Cell Death

Apoptosis of retinal neurons is accelerated by diabetes and the onset of increased cell death occurs early in disease progression. The present study was designed to evaluate molecular defects that contribute to diabetes-induced neuroretinal apoptosis and visual dysfunction. We recently demonstrated that the stress response protein REDD1 is induced by hyperglycemia in the retina of diabetic mice in a manner that contributes to neuronal cell death and visual dysfunction. One potential mechanism whereby REDD1 promotes neuronal apoptosis is through upregulation of reactive oxygen species (ROS), as the protein forms a pro-oxidant complex. In the retina of streptozotocin-induced diabetic mice, ROS levels, neuronal cell death, and REDD1 expression were increased concomitant with attenuation of contrast sensitivity thresholds, as assessed by behavioral optomotry. In R28 retinal cells in culture, hyperglycemic conditions increased REDD1 expression, ROS levels, and cell death. However, addition of the antioxidant N-acetyl-L-cysteine (NAC) to culture medium was sufficient to prevent the increase in ROS and cell death in response to hyperglycemic conditions. Moreover, REDD1-deficient cells did not exhibit increased ROS or cell death in response to hyperglycemic conditions. Similarly, in REDD1-deficient diabetic mice, ROS levels and cell death in retina were similar to that observed in nondiabetic mice. Finally, oral administration of NAC improved contrast sensitivity in diabetic mice relative to vehicle treated diabetic controls, demonstrating a key role for ROS levels in diabetes-induced visual dysfunction. Overall, the findings support therapeutic approaches targeting REDD1 and ROS production in the prevention of diabetes-induced visual dysfunction. Disclosure W.P. Miller: None. A. Toro: None. M.D. Dennis: None.

Nrf2 protects against diabetic dysfunction of endothelial progenitor cells via regulating cell senescence

Diabetes is associated with an increased risk of cardio-vascular disease. A decrease in the number and functionality of endothelial progenitor cells (EPCs) leads to reduced endothelial repair and the development of cardiovascular disease. The aim of the present study was to explore the effect and underlying mechanisms of nuclear factor erythroid 2-related factor 2 (Nrf2) on EPC dysfunction caused by diabetic mellitus. The biological functions of EPCs in streptozotocin-induced diabetic mice were evaluated, including migration, proliferation, angiogenesis and the secretion of vascular endothelial growth factor (VEGF), stromal-derived growth factor (SDF) and nitric oxide (NO). Oxidative stress levels in diabetic EPCs were also assessed by detecting intracellular reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA). EPC senescence was evaluated by measuring p16 and b-gal expression and observing the senescence-associated secretory phenotype. In addition, the function of EPCs and level of oxidative stress were assessed following Nrf2 silencing or activation. Nrf2 silencing resulted in a decrease of EPC biological functions, accelerated cell senescence and increased oxidative stress, as indicated by ROS and MDA upregulation accompanied with decreased SOD activity. Furthermore, Nrf2 silencing inhibited migration, proliferation and secretion in EPCs, while it increased oxidative stress and cell senescence. Nrf2 activation protected diabetic EPCs against the effects of oxidative stress and cell senescence, ameliorating the biological dysfunction of EPCs derived from mice with diabetes. In conclusion, Nrf2 overexpression protected against oxidative stress-induced functional damage in EPCs derived from diabetic mice by regulating cell senescence.

Bioengineered Boronic Ester Modified Dextran Polymer Nanoparticles as Reactive Oxygen Species Responsive Nanocarrier for Ischemic Stroke Treatment.

Ischemic stroke is a leading cause of long-term disability and death worldwide. Current drug delivery vehicles for the treatment of ischemic stroke are less than satisfactory, in large part due to their short circulation lives, lack of specific targeting to the ischemic site, and poor controllability of drug release. In light of the upregulation of reactive oxygen species (ROS) in the ischemic neuron, we herein developed a bioengineered ROS-responsive nanocarrier for stroke-specific delivery of a neuroprotective agent, NR2B9C, against ischemic brain damage. The nanocarrier is composed of a dextran polymer core modified with ROS-responsive boronic ester and a red blood cell (RBC) membrane shell with stroke homing peptide (SHp) inserted. These targeted "core-shell" nanoparticles (designated as SHp-RBC-NP) could thus have controlled release of NR2B9C triggered by high intracellular ROS in ischemic neurons after homing to ischemic brain tissues. The potential of the SHp-RBC-NP for ischemic stroke therapy was systematically evaluated in vitro and in rat models of middle cerebral artery occlusion (MCAO). In vitro results showed that the SHp-RBC-NP had great protective effects on glutamate-induced cytotoxicity in PC-12 cells. In vivo pharmacokinetic (PK) and pharmacodynamic (PD) testing further demonstrated that the bioengineered nanoparticles can drastically prolong the systemic circulation of NR2B9C, enhance the active targeting of the ischemic area in the MCAO rats, and reduce ischemic brain damage.

MP54-09 TM4SF1 IS A NOVEL BIOMARKER FOR HUMAN MUSCLE INVASIVE BLADDER CANCER (MIBC)

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology I1 Apr 2018MP54-09 TM4SF1 IS A NOVEL BIOMARKER FOR HUMAN MUSCLE INVASIVE BLADDER CANCER (MIBC) Rui Cao, Gang Wang, Kaiyu Qian, Liang Chen, Yu Xiao, and Xinghuan Wang Rui CaoRui Cao More articles by this author , Gang WangGang Wang More articles by this author , Kaiyu QianKaiyu Qian More articles by this author , Liang ChenLiang Chen More articles by this author , Yu XiaoYu Xiao More articles by this author , and Xinghuan WangXinghuan Wang More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1700AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES BCa is a heterogeneous disease: non-muscle-invasive bladder cancer (NMIBC) that usually recur but rarely process, and muscle-invasive bladder cancer (MIBC) that progress quickly. Recent studies identify numerous clinical variables and molecular biomarkers, such as chromosomal markers, genetic variations or epigenetic alterations for prediction of its occurrence and recurrence. However, effective biomarkers to distinguish MIBC from NMIBC are urgently needed. Transmembrane-4-L-Six-Family-1 (TM4SF1) is a member of L6 family, which functions as signal transducer to regulate cell development, growth and motility. The aim of the study is to investigate the TM4SF1 as the specific biomarker of MIBC via bioinformatics analysis. METHODS Through mining the big data from GEO and TCGA, a specific MIBC differentially expressed gene (DEG) called TM4SF1 was screened by R and validated by oncomine database, etc. Then qRT-PCR, immunofluorescence and immunohistochemistry analysis were used to detect the transcription and translation level of TM4SF1 in bladder tissues. Clinical correlation was measured by connecting the expression of TM4SF1 with patients′ information. Moreover, TM4SF1 knockdown and overexpression BCa cell model was established by gene regulation. Then flow cytometry, CCK-8 assay, western blot, etc. were used to detect cell cycle distribution, cell growth and related protein expression in vitro. Furthermore, the xenograft model was used to measure the tumor growth of BCa cells transplantation in vivo. RESULTS Our results suggested that TM4SF1 was strongly upregulated in MIBC tissues, corroborated with transcriptome analysis. Moreover, tissue microarray (TMA) showed significant correlations between TM4SF1 with T stage, TNM stage, lymph node metastasis status and survival rate of MIBC patient. Furthermore, in vitro and in vivo studies suggested that proliferation of human bladder cancer (BCa) cells was significantly suppressed by downregulated TM4SF1. Functionally, reduction of TM4SF1 could induce cell cycle arrest and apoptosis possibly via upregulation of reactive oxygen species (ROS) in the BCa cells. Interestingly, these observations were scavenged by treatment of GW9662 (antagonist of PPARγ) and resveratrol (activator of SIRT1), respectively. CONCLUSIONS We have first shown tumor promoting effect of TM4SF1 on MIBC. TM4SF1 could be a biomarker to predict the survival rate of MIBC patients and malignancy of the tumor. And our study suggest that the regulation of ROS metabolism and cell cycle for TM4SF1 in BCa cells was mediated by PPARγ-SIRT1 feedback loop. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e715 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Rui Cao More articles by this author Gang Wang More articles by this author Kaiyu Qian More articles by this author Liang Chen More articles by this author Yu Xiao More articles by this author Xinghuan Wang More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Anesthetic Agents and Neuronal Autophagy. What We Know and What We Don't.

Ethanol is known to have both γ-Aminobutyric acid agonist and Nmethyl- D-aspartate antagonist characteristics similar to commonly used volatile anesthetic agents. Recent evidence demonstrates that autophagy can reduce the development of ethanol induced neurotoxicity. Recent studies have found that general anesthesia can cause longterm impairment of both mitochondrial morphogenesis and synaptic transmission in the developing rat brain, both of which are accompanied by enhanced autophagy activity. Autophagy may play an important role in general anesthetic mediated neurotoxicity. This review outlines the role of autophagy in the development of anesthetic related neurotoxicity and includes an explanation of the role of autophagy in neuronal cell survival and death, the relationship between anesthetic agents and neuronal autophagy, possible molecular and cellular mechanisms underlying general anesthetic agent induced activation of neuronal autophagy in the developing brain, and potential therapeutic approaches aimed at modulating autophagic pathways. In a time- and concentration-dependent pattern, general anesthetic agents can disrupt intracellular calcium homeostasis which enhances both autophagy and apoptosis activation. The degree of neural cell injury may be ultimately determined by the interplay between autophagy and apoptosis. It appears likely that the increase in calcium flux associated with some anesthetic agents disrupts lysosomal function. This results in an over-activation of endosomal- lysosomal trafficking causing mitochondrial damage, reactive oxygen species upregulation, and lipid peroxidation. Autophagy may play a role in the development of anesthetic related neurotoxicity. Understanding this may lead to strategies or therapies aimed at preventing or ameliorating general anesthetic agent mediated neurotoxicity.

Curcumin targets multiple enzymes involved in the ROS metabolic pathway to suppress tumor cell growth

Curcumin has been reported to exhibit anti-tumorigenic activity; however, since its precise actions remain unclear, its effects are considered to be deceptive. In the present study, we confirmed the anti-tumorigenic effects of curcumin on CML-derived leukemic cells in a xenograft model and in vitro culture system. In vitro pull-down and mass analyses revealed a series of enzymes (carbonyl reductase, glutathione-S-transferase, glyoxalase, etc.) that function in a reactive oxygen species (ROS) metabolic pathway as curcumin-binding targets, the expression of which was up-regulated in human leukemia. Curcumin increased ROS levels over the threshold in leukemic cells, and the antioxidant, glutathione (GSH) and overexpression of curcumin-binding enzymes partially mitigated the up-regulation of ROS and growth inhibition caused by curcumin. These results show that curcumin specifically inhibits tumor growth by increasing ROS levels over the threshold through the miscellaneous inhibition of ROS metabolic enzymes. Curcumin has potential in therapy to regulate ROS levels in tumor cells, thereby controlling tumor growth.

Abstract TP252: MicroRNA-376a-5p Improves Outcome for Acute Ischemic Stroke After Endovascular Mechanical Reperfusion in Rat Model

Background: MicroRNAs are involved in regulating cerebral ischemia process, but its underlying mechanism is unclear. The removal of intraluminal filament in the transient middle cerebral artery occlusion (tMCAO) model resulted in instantaneous recanalization, similar to endovascular thrombectomy in stroke. We previously showed microRNA-376a down-regulation in the ischemic brain in rat tMCAO model with hemorrhagic transformation. This study investigated the role of miR-376a-5p in cerebral ischemia and reperfusion injury after mechanical reperfusion. Methods: The intraluminal filament tMCAO model was established in rats with 2 hour ischemia followed by reperfusion. The expression of miR-376a-5p in the ischemic brain and blood were quantified. Intravenous miR-376a-5p agomir was delivered before reperfusion or after reperfusion. Infarct volume, brain water content, neurological score, blood-brain barrier (BBB) damage, and levels of miR-376a-5p, NADPH oxidase (NOX) 4, Caspase-3 and aquaporin-4 were determined at 24-hour after ischemia. Results: MiR-376a-5p levels in the infarction core and peri-infarct cortex were decreased at 3 hours after reperfusion in tMCAO model. The decreased levels of miR-376a in ischemic brain and blood lasted for 24 hours after ischemia. Intravenous miR-376a-5p agomir before and after reperfusion increased the expression of miR-376a and suppressed the up-regulation of NOX4, Caspase-3 and AQP-4 in the infarction core and peri-infarct cortex compared with the tMCAO group (p&lt;0.01). Dual-luciferase reporter system showed that NOX4, Caspase-3, and AQP-4 were the direct target genes of miR-376a-5p. MiR-376a overexpression inhibited the up-regulation of reactive oxygen species and neuronal apoptosis in the peri-infarct cortex. Intravenous miR-376a agomir reduced infarct volume, brain edema and reperfusion-induced BBB breakdown at 24-hour after ischemia, resulting in improved neurological outcome (p&lt;0.01). Conclusions: MiR-376a-5p overexpression protects against cerebral ischemia and reperfusion injury via downregulating NOX4, Caspase-3 and AQP-4. Infusion of miR-376a-5p agomir represents a novel adjunctive therapeutic strategy to improve outcome after mechanical reperfusion for acute stroke.

Oxidative stress regulates cellular bioenergetics in esophageal squamous cell carcinoma cell.

The aim of the present study was to explore the effects of oxidative stress induced by CoCl2 and H2O2 on the regulation of bioenergetics of esophageal squamous cell carcinoma (ESCC) cell line TE-1 and analyze its underlying mechanism. Western blot results showed that CoCl2 and H2O2 treatment of TE-1 cells led to significant reduction in mitochondrial respiratory chain complex subunits expression and increasing intracellular reactive oxygen species (ROS) production. We further found that TE-1 cells treated with CoCl2, a hypoxia-mimicking reagent, dramatically reduced the oxygen consumption rate (OCR) and increased the extracellular acidification rate (ECAR). However, H2O2 treatment decreased both the mitochondrial respiration and aerobic glycolysis significantly. Moreover, we found that H2O2 induces apoptosis in TE-1 cells through the activation of PARP, Caspase 3, and Caspase 9. Therefore, our findings indicate that CoCl2 and H2O2 could cause mitochondrial dysfunction by up-regulation of ROS and regulating the cellular bioenergy metabolism, thus affecting the survival of tumor cells.