Showing papers in "Cell Biochemistry and Function in 2016"
TL;DR: This work has shown that novel systems provide a huge potential for large animal models to investigate the regulatory factors and molecular pathways that contribute to CVD in vivo and will help bridge the gap between basic science and clinical applications by facilitating the refinement of therapies for cardiovascular disease.
Abstract: The human cardiovascular system is a complex arrangement of specialized structures with distinct functions. The molecular landscape, including the genome, transcriptome and proteome, is pivotal to the biological complexity of both normal and abnormal mammalian processes. Despite our advancing knowledge and understanding of cardiovascular disease (CVD) through the principal use of rodent models, this continues to be an increasing issue in today's world. For instance, as the ageing population increases, so does the incidence of heart valve dysfunction. This may be because of changes in molecular composition and structure of the extracellular matrix, or from the pathological process of vascular calcification in which bone-formation related factors cause ectopic mineralization. However, significant differences between mice and men exist in terms of cardiovascular anatomy, physiology and pathology. In contrast, large animal models can show considerably greater similarity to humans. Furthermore, precise and efficient genome editing techniques enable the generation of tailored models for translational research. These novel systems provide a huge potential for large animal models to investigate the regulatory factors and molecular pathways that contribute to CVD in vivo. In turn, this will help bridge the gap between basic science and clinical applications by facilitating the refinement of therapies for cardiovascular disease.
99 citations
TL;DR: Preliminary findings indicate that downregulated serum miR‐221 might be a potential biomarker for PD evaluation.
Abstract: Parkinson's disease (PD) is the common age-related neurodegenerative disorder. Sensitive, noninvasive biomarkers that facilitate PD diagnosis and stage assignment are currently unavailable. This study aims to investigate the potential of 16 previous reported PD-associated miRNAs as novel biomarkers for PD. The expression of 16 serum miRNAs was measured by quantitative reverse transcriptase polymerase chain reaction in 138 PD patients and 112 control populations. Analyses were undertaken to assess the specificity and sensitivity of miRNAs to predict PD. In addition, the relationship between deregulated miRNAs and Part III of the United Parkinson's Disease Rating Scale (UPDRS-III) and Part V of the UPDRS (UPDRS-V; the modified Hoehn and Yahr staging of PD) in PD patients was also assessed. It was found that the serums miR-29c, miR-146a, miR-214, and miR-221 were significantly decreased in PD patients compared with healthy control populations. In addition, serum miR-221 was positively correlated with UPDRS-III (r = .4702) and UPDRS-V (r = .4788) score in PD patients. Furthermore, the receiver operating characteristic result of serum miR-221 for prediction of PD was 0.787. Our preliminary findings indicate that downregulated serum miR-221 might be a potential biomarker for PD evaluation.
82 citations
TL;DR: It is shown that obesity is associated with increased risk of several cancer types, including colon, breast, endometrium, liver, kidney, esophagus, gastric, pancreatic, gallbladder, and leukemia, and can also lead to poorer treatment.
Abstract: Nowadays, obesity is considered as a serious and growing global health problem. It is documented that the overweight and obesity are major risk factors for a series of noncommunicable diseases, and in recent years, the obesity-cancer link has received much attention. Numerous epidemiological studies have shown that obesity is associated with increased risk of several cancer types, including colon, breast, endometrium, liver, kidney, esophagus, gastric, pancreatic, gallbladder, and leukemia, and can also lead to poorer treatment.
We review here the epidemiological and experimental evidences for the association between obesity and cancer. Specifically, we discuss potential mechanisms focusing how dysfunctional angiogenesis, chronic inflammation, interaction of proinflammatory cytokines, endocrine hormones, and adipokines including leptin, adiponectin insulin, growth factors, estrogen, and progesterone and strikingly, cell metabolism alteration in obesity participate in tumor development and progression, resistance to chemotherapy, and targeted therapies such as antiangiogenic and immune therapies.
77 citations
TL;DR: Viruses from the Filoviridae family, as many other virus families, require an acidic pH for successful infection and are therefore susceptible to the actions of 4‐aminoquinolines, such as chloroquine.
Abstract: Viruses from the Filoviridae family, as many other virus families, require an acidic pH for successful infection and are therefore susceptible to the actions of 4-aminoquinolines, such as chloroquine. Although the mechanisms of action of chloroquine clearly indicate that it might inhibit filoviral infections, several clinical trials that attempted to use chloroquine in the treatment of other acute viral infections - including dengue and influenza A and B - caused by low pH-dependent viruses, have reported that chloroquine had no clinical efficacy, and these results demoted chloroquine from the potential treatments for other virus families requiring low pH for infectivity. The present review is aimed at investigating whether chloroquine could combat the present Ebola virus epidemic, and also at exploring the main reasons for the reported lack of efficacy. Literature was sourced from PubMed, Scopus, Google Scholar, reference list of articles and textbooks - Fields Virology (Volumes 1and 2), the cytokine handbook, Pharmacology in Medicine: Principles and Practice, and hydroxychloroquine and chloroquine retinopathy. The present analysis concludes that (1) chloroquine might find a place in the treatment of Ebola, either as a monotherapy or in combination therapies; (2) the ineffectiveness of chloroquine, or its analogue, hydroxychloroquine, at treating infections from low pH-dependent viruses is a result of the failure to attain and sustain a steady state concentration sufficient to increase and keep the pH of the acidic organelles to approximately neutral levels; (3) to successfully treat filoviral infections - or other viral infections that emerge or emerged from low pH-dependent viruses - a steady state chloroquine plasma concentration of at least 1 µg/mL(~3.125 μM/L) or a whole blood concentration of 16 μM/L must be achieved and be sustained until the patients' viraemia becomes undetectable. These concentrations, however, do not rule out the efficacy of other, higher, steady state concentrations - although such concentrations might be accompanied by severe adverse effects or toxicities. The feasibility of the conclusion in the preceding texts has recently been supported by a subsequent study that shows that amodiaquine, a derivative of CQ, is able to protect humans infected with Ebola from death.
76 citations
TL;DR: The preliminary findings indicate that serum miR‐223 might be a potential biomarker for AD evaluation, and the combination of miRNAs gave improved sensitivity/specificity for AD prediction than either miRNA alone.
Abstract: Alzheimer's disease (AD) is an age-related neurodegenerative disorder that is characterized by progressive memory loss and deteriorated higher cognitive functions. An economical, rapid and noninvasive biomarker for AD has not been identified. We aimed to investigate the diagnostic value of serum miR-223 and miR-519 in AD. The expressions of miR-223 and miR-519, with previously reported AD-associated miR-29 and miR-125b, were measured by quantitative reverse transcription polymerase chain reaction in the serum of 84 probable sporadic AD patients (age onset > 65 years) and 62 healthy control populations in China. Analyses were undertaken to assess the specificity and sensitivity of miRNAs to predict AD. In addition, the relationship between miRNAs and mini mental state examination (MMSE) scores in AD patients was also assessed. Serum miR-29, miR-125b and miR-223 were significantly decreased, but serum miR-519 was significantly increased in AD patients compared with healthy blood donors. In addition, serum miR-223 was strongly positively correlated with MMSE score in AD patients but serum miR-519 was not. Importantly, the receiver operating characteristic (ROC) result of serum miR-223 for prediction of AD was 0.786, higher than those of serum miR-29 (0.734) or miR-125b (0.726). The combination of serum miR-223 and miR-125b gave improved sensitivity/specificity for AD prediction (area under the ROC curve, 0.879) than either miRNA alone. Our preliminary findings indicate that serum miR-223 might be a potential biomarker for AD evaluation. Copyright © 2016 John Wiley & Sons, Ltd.
67 citations
TL;DR: The regulation and dysregulation of DNA methylation in plants and mammals is discussed and the similarities and differences between mammals and plants are discussed.
Abstract: DNA methylation is a major epigenetic marking mechanism regulating various biological functions in mammals and plant. The crucial role of DNA methylation has been observed in cellular differentiation, embryogenesis, genomic imprinting and X-chromosome inactivation. Furthermore, DNA methylation takes part in disease susceptibility, responses to environmental stimuli and the biodiversity of natural populations. In plant, different types of environmental stress have demonstrated the ability to alter the archetype of DNA methylation through the genome, change gene expression and confer a mechanism of adaptation. DNA methylation dynamics are regulated by three processes de novo DNA methylation, methylation maintenance and DNA demethylation. These processes have their similarities and differences between mammals and plants. Furthermore, the dysregulation of DNA methylation dynamics represents one of the primary molecular mechanisms of developing diseases in mammals. This review discusses the regulation and dysregulation of DNA methylation in plants and mammals. Copyright © 2016 John Wiley & Sons, Ltd.
55 citations
TL;DR: Findings suggest that plasmamiR‐92a, miR‐100 and miR-143 could be promising novel circulating biomarkers in clinical detection of BC.
Abstract: The application of microRNAs (miRNAs) as potential biomarkers and therapy targets has been widely investigated in many kinds of cancers. Recent advantages of serum miRNAs open a new realm of possibilities for non-invasive diagnosis and prognosis of bladder cancer (BC). The aim of our study was to identify plasma miR-92a, miR-100 and miR-143 expression signatures in patients with BC to introduce new markers for establishing BC diagnosis and prognosis. Blood samples were collected from 70 BC patients and 62 controls. An expression of three target miRNAs (miR-92a, miR-100 and miR-143) was measured using quantitative real-time PCR method. Results were correlated with clinicopathological data and analysed. Plasma levels of miR-92a, miR-100 and miR-143 were significantly lower in BC patients than in control group. Receiver operator characteristic analysis revealed that the sensitivity and specificity values of miR-92a were 97·1% and 76·7%, respectively, with a cut-off value of 0·573. The sensitivity and specificity values of miR-100 were 90% and 66·7%, respectively, with a cut-off value of 0·644. The sensitivity and specificity values of miR-143 were 78·6% and 93·3%, respectively, with a cut-off value of 0·164. This study explores the existence of specific plasma miRNAs as early diagnostic biomarkers for BC in Egyptian patients; and these findings suggest that plasma miR-92a, miR-100 and miR-143 could be promising novel circulating biomarkers in clinical detection of BC.
48 citations
TL;DR: It is demonstrated that Trpv1, a capsaicin receptor, is expressed in brown adipose tissue, and that its expression level is increased during the differentiation of HB2 brown preadipocytes.
Abstract: The ingestion of capsaicin, the principle pungent component of red and chili peppers, induces thermogenesis, in part, through the activation of brown adipocytes expressing genes related to mitochondrial biogenesis and uncoupling such as peroxisome proliferator-activated receptor (Ppar) γ coactivator-1α (Pgc-1α) and uncoupling protein 1 (Ucp1). Capsaicin has been suggested to induce the activation of brown adipocytes, which is mediated by the stimulation of sympathetic nerves. However, capsaicin may directly affect the differentiation of brown preadipocytes, brown adipocyte function, or both, through its significant absorption. We herein demonstrated that Trpv1, a capsaicin receptor, is expressed in brown adipose tissue, and that its expression level is increased during the differentiation of HB2 brown preadipocytes. Furthermore, capsaicin induced calcium influx in brown preadipocytes. A treatment with capsaicin in the early stage of brown adipogenesis did not affect lipid accumulation or the expression levels of Fabp4 (a gene expressed in mature adipocytes), Pparγ2 (a master regulator of adipogenesis) or brown adipocyte-selective genes. In contrast, a treatment with capsaicin in the late stage of brown adipogenesis slightly increased the expression levels of Fabp4, Pparγ2 and Pgc-1α. Although capsaicin did not affect the basal expression level of Ucp1, Ucp1 induction by forskolin was partially inhibited by capsaicin, irrespective of the dose of capsaicin. The results of the present study suggest the direct effects of capsaicin on brown adipocytes or in the late stage of brown adipogenesis.
41 citations
TL;DR: The results show overwhelming evidence corroborating the suppressive effect of miR‐29b on TGF‐β1‐induced LX‐2 cell activation and revealing the existence of crosstalk between miR-29b and TGF-β1/Smad3 during LX‐ 2 activation, suggesting a feedback loop betweenmiR‐ 29b andTGF‐ β1/ Smad3 signalling that promotes liver fibrosis.
Abstract: The microRNA (miR)-29 family is closely associated with fibrotic processes by virtue of its low expression in many tissues during organ fibrosis The present study investigated whether miR-29b overexpression suppressed hepatic stellate cell (HSC) activation and its interactions with transforming growth factor (TGF)-β1/mothers against decapentaplegic homolog 3 (Smad3), a classical signal transduction pathway contributing to the activation of HSCs The results showed that transfection of LX-2 (human HSC) cells with miR-29b mimic or pSUPER-Smad3 silencing (si)RNA resulted in significantly increased expression of miR-29b and decreased expression of Smad3 miR-29b overexpression inhibited proliferation of LX-2 cells 24 h after transfection Both miR-29b overexpression and Smad3 silencing antagonized the effects of TGF-β1 on the expression of α-smooth muscle actin (α-SMA) and collagen type I (col-1) Furthermore, infection with miR-29b mimics suppressed Smad3 and TGF-β1 expression, suggesting that miR-29b inhibited LX-2 activation mediated by both Smad3 and TGF-β1 Nevertheless, primary miR-29a/b1, miR-29b2/c and mature miR-29b were downregulated by TGF-β1 and stimulated by Smad3 silencing, suggesting that TGF-β1/Smad3 signalling pathway regulate not just mature miR-29b but also its transcription In summary, our results show overwhelming evidence corroborating the suppressive effect of miR-29b on TGF-β1-induced LX-2 cell activation The results also revealed the existence of crosstalk between miR-29b and TGF-β1/Smad3 during LX-2 activation, suggesting a feedback loop between miR-29b and TGF-β1/Smad3 signalling that promotes liver fibrosis Copyright © 2016 John Wiley & Sons, Ltd
33 citations
TL;DR: Overexpression of miR‐204‐5p and SIRT1 in human HCC cell lines caused the suppression of cell survival ability, the increase of apoptosis, and drug sensitivity, and results indicate that miR-204-5p may play an important role in the development of HCC.
Abstract: Hepatocellular carcinoma (HCC) is the most common type of cancer, which presents rapid tumor growth, drug resistance, and metastasis. Recently, microRNAs are shown to be involved in the cell biological processes in HCC, but the underlying molecular mechanisms remain unclear. This study aimed to investigate the cellular function and molecular mechanism of miR-204-5p in HCC. SIRT1 mRNA and miR-204-5p were examined by real-time reverse transcription polymerase chain reaction. SIRT1 protein levels were measured by Western blotting. Cell proliferation assay was performed to confirm colony formation. Invasion assay was performed by transwell system. SPSS 15.0 for Windows was used for statistical analysis. SIRT1 was a potential oncogene in cancer, which was identified as a direct target of miR-204-5p. Overexpression of miR-204-5p in human HCC cell lines (BEL-7405 and QGY-7701) caused the suppression of cell survival ability, the increase of apoptosis, and drug sensitivity. SIRT1 was overexpressed in human HCC tissues and was negatively related to miR-204-5p levels. These results indicate that miR-204-5p and SIRT1 may play an important role in the development of HCC.
31 citations
TL;DR: The data suggest that aHIF overexpression can be used as a potential biomarker in breast cancer, however, further studies are needed for the evaluation of its mechanism of action in regulation of HIF‐1α expression in different pathological conditions.
Abstract: Hypoxia-inducible factors (HIFs) have been shown to be upregulated in tumor tissues and linked with tumor progression and metastasis in breast cancer. Among regulatory mechanisms for HIF expression is a natural occurring antisense named aHIF, which has been shown to be overexpressed in breast cancer and influence the level of the HIF-1α transcript. In the present study, we analyzed the expression of HIF-1α and aHIF in breast cancer tissues versus adjacent noncancer tissues (ANCTs) in relation with the clinical and biological behavior of the tumors. aHIF has been shown to be expressed in 67.4% of invasive ductal carcinoma samples, while none of ANCTs showed its expression. HIF-1α has been expressed in all of tumors and 90% of ANCTs. Comparison of HIF-1α expression level between tumor and ANCT tissues showed a total upregulation in tumor samples. No statistically significant association has been found between the level of HIF-1α expression in tumor samples and clinicopathologic and demographic characteristics such as age, tumor size, estrogen receptor status, progesterone receptor status, HER2/neu expression level, lymph node status, histological grade, and stage except for a weak correlation between HIF-1α expression and Ki-67 status. Besides, we could not detect any significant correlation between relative expression of HIF-1α and aHIF in tumor samples. Collectively, these data suggest that aHIF overexpression can be used as a potential biomarker in breast cancer. However, further studies are needed for the evaluation of its mechanism of action in regulation of HIF-1α expression in different pathological conditions. HIF-1α overexpression results in the upregulation of several genes that participated in cancer-associated pathways such as proliferation, angiogenesis, and glucose metabolism. We showed that HIF-1α is upregulated in breast tumor samples compared with adjacent noncancerous tissues. Its expression has been associated with Ki-67 status. Its natural occurring antisense is only expressed in tumor tissues. Thus, it can be used as a potential biomarker in breast cancer.
TL;DR: Results indicated that PlGF knockdown decreased migration and induced apoptosis through PI3K/Akt1 and p38MAPK signal transduction in AGS cells.
Abstract: The molecular signalling of placental growth factor (PlGF), a member of the vascular endothelial growth factor family, was not uncovered in human adenocarcinoma gastric cell line (AGS). The purpose of this study was to examine the inhibitory effects of PlGF knockdown on cell proliferation, apoptosis and migration through p38 mitogen-activated protein kinase (p38MAPK) and PI3K pathways in human adenocarcinoma gastric cell line (AGS). To study PlGF knockdown effect, AGS cells were treated with 40 pmol of small interfering RNA (siRNA) related to PlGF gene and also a scrambled siRNA as control. Trypan Blue and Anexin V staining of AGS cells treated with PlGF-specific siRNA showed induction of apoptosis. Wound healing assay and zymography indicated that cellular migration and matrix metalloproteinases activities were reduced in response to PlGF knockdown. Phosphorylation of Akt and p38MAPK was reduced in AGS cells treated with PlGF-specific siRNA. PlGF knockdown decreased transcripts of PI3K, Akt, p38MAPK, PCNA, Caspase-3, OCT3/OCT4 and CD44, but elevated p53 and SOX2 transcripts. Our results indicated that PlGF knockdown decreased migration and induced apoptosis through PI3K/Akt1 and p38MAPK signal transduction in AGS cells. Copyright © 2016 John Wiley & Sons, Ltd.
TL;DR: It is suggested that vitamin D may offer a therapeutic potential for patients with PD by evaluating the anti‐oxidative stress and anti‐inflammatory properties of the active form of vitamin D3, 1,α, 25‐dihydroxyvitamin D3 in a mouse cone cell line, 661W.
Abstract: Photoreceptor degeneration (PD) refers to a group of heterogeneous outer retinal dystrophies characterized by the death of photoreceptors. Both oxidative stress and inflammation are involved in the pathogenesis of PD. We investigate whether vitamin D has a potential for the treatment of PD by evaluating the anti-oxidative stress and anti-inflammatory properties of the active form of vitamin D3 , 1,α, 25-dihydroxyvitamin D3 , in a mouse cone cell line, 661W. Mouse cone cells were treated with H2 O2 or a mixture of H2 O2 and vitamin D; cell viability was determined. The production of reactive oxygen species (ROS) in treated and untreated cells was measured. The expression of key anti-oxidative stress and inflammatory genes in treated and untreated cells was determined. Treatment with vitamin D significantly increased cell viability and decreased ROS production in 661W cells under oxidative stress induced by H2 O2 . H2 O2 treatment in 661W cells can significantly down-regulate the expression of antioxidant genes and up-regulate the expression of neurotoxic cytokines. Vitamin D treatment significantly reversed these effects and restored the expression of antioxidant genes. Vitamin D treatment also can block H2 O2 induced oxidative damages. The data suggested that vitamin D may offer a therapeutic potential for patients with PD.
TL;DR: The in vitro anticancer activity of C‐ring modified semi‐synthetic derivative of colchicine and 4h suggest that 4h has better selectivity and is a microtubule depolymerizer, which activates dual cell‐death machineries, and thus, it could be a potential novel therapeutic agent in cancer therapy.
Abstract: Colchicine is a tubulin-binding natural product isolated from Colchicum autumnale. Here we report the in vitro anticancer activity of C-ring modified semi-synthetic derivative of colchicine; N-[(7S)-1,2,3-trimethoxy-9-oxo-10-(4-phenyl-piperidin-1-yl)-5,6,7,9 tetrahydrobenzo[a]heptalen-7-yl]acetamide (4h) on colon cancer HCT-116 cell line. The compound 4h was screened for anti-proliferative activity against different human cancer cell lines and was found to exhibit higher cytotoxicity against colon cancer cell lines HCT-116 and Colo-205 with IC50 of 1 and 0.8 μM respectively. Cytotoxicity of the compound to the normal fR2 breast epithelial cells and normal HEK293 human embryonic kidney cells was evaluated in concentration and time-dependent manner to estimate its selectivity for cancer cells which showed much better selectivity than that of colchicine. Compound 4h induced cell death in HCT-116 cells by activating apoptosis and autophagy pathways. Autophagy inhibitor 3-MA blocked the production of LC3-II and reduced the cytotoxicity in response to 4h, but did not affect apoptosis, suggesting thereby that these two were independent events. Reactive oxygen species scavenger ascorbic acid pretreatment not only decreased the reactive oxygen species level but also reversed 4h induced cytotoxicity. Treatment with compound 4h depolymerized microtubules and the majority of cells arrested at the G2/M transition. Together, these data suggest that 4h has better selectivity and is a microtubule depolymerizer, which activates dual cell-death machineries, and thus, it could be a potential novel therapeutic agent in cancer therapy.
TL;DR: It is demonstrated that miR‐148a‐3p may regulate RUNX3 expression through the modulation of DNMT1‐dependent DNA methylation in LSCC, providing a novel target and a potential therapeutic pathway against LSCC.
Abstract: Laryngeal squamous cell carcinoma (LSCC) is a highly aggressive malignant cancer and accounts for 1% to 2% of all malignancies diagnosed worldwide. Runt-related transcription factor 3 (RUNX3), an important tumor suppressor, is known to related to lymph node metastasis and the development of LSCC. However, the biological roles and potential mechanisms RUNX3 expression was not well understood. In this study, we reported that the RUNX3 was significantly downregulated and highly methylated in LSCC compared with their matched normal. The enforced expression of RUNX3 inhibited LSCC cell migration, invasion, and proliferation, whereas the inhibition of RUNX3 did the opposite. We identified that RUNX3 was regulated by miR-148a-3p and found that the expression level of miR-148-3p was significantly decreased and positively related with the expression of RUNX3 in LSCC. We also identified that DNA methyltransferase enzyme DNA (cytosine-5-)-methyltransferase 1 (DNMT1) was targeted by miR-148a-3p in LSCC. The knockdown of DNMT1 promoted the expression of RUNX3 and inhibited migration, invasion, and proliferation in LSCC cells. In summary, our study demonstrated that miR-148a-3p may regulate RUNX3 expression through the modulation of DNMT1-dependent DNA methylation in LSCC, providing a novel target and a potential therapeutic pathway against LSCC. LSCC is a highly aggressive malignant cancer and accounts for 1% to 2% of all malignancies diagnosed worldwide. In this study, we reported that RUNX3, an important tumor suppressor, was significantly downregulated and highly methylated in LSCC compared with their matched normal. The overexpression of RUNX3 inhibited LSCC cell migration, invasion, and proliferation, whereas the inhibition of RUNX3 did the opposite. Moreover, RUNX3 was regulated by miR-148a-3p, which targeted DNA methyltransferase enzyme DNMT1 in LSCC cells. Therefore, miR-148a-3p may regulate RUNX3 expression through the modulation of DNMT1-dependent DNA methylation in LSCC, providing a novel target and a potential therapeutic pathway against LSCC.
TL;DR: The latest developments in understanding of PEBP1 function in maintaining cell integrity are outlined and many drugs that mainly target the interaction ofPEBP1 with its partners are identified.
Abstract: UNLABELLED Phosphatidylethanolamine-binding protein 1 (PEBP1), also known as Raf kinase inhibitor protein, belongs to PEBP family of proteins. It is known to interact with many proteins that are mainly involved in pathways that monitor cell proliferation and differentiation. PEBP1 in many cells interacts with several pathways, namely MAPK, GRK2, NF-кB, etc. that keeps the cell proliferation and differentiation in check. This protein is expressed by many cells in humans, including neurons where it is predominantly involved in production of choline acetyltransferase. Deregulated PEBP1 is known to cause cancer, diabetic nephropathy and neurodegenerative diseases like Alzheimer's and dementia. Recent research led to the discovery of many drugs that mainly target the interaction of PEBP1 with its partners. These compounds are known to bind PEBP1 in its conserved domain which abrogate its association with interacting partners in several different pathways. We outline here the latest developments in understanding of PEBP1 function in maintaining cell integrity. Copyright © 2016 John Wiley & Sons, Ltd. SIGNIFICANCE OF THE STUDY Phosphatidylethanolamine-binding protein is crucial in regulation of MAPK and PKC pathways. Its diverse roles, including regulating these pathways keep cell differentiation and proliferation in check. This review outlines some latest findings which greatly add to our current knowledge of phosphatidylethanolamine-binding protein.
TL;DR: This study suggests that glyoxalase II, through a specific interaction of its catalytic site with target proteins, could be able to perform a rapid and specific protein S‐glutathionylation using its natural substrate S‐d‐lactoyl glutathione.
Abstract: Glyoxalase II, the second of 2 enzymes in the glyoxalase system, is a hydroxyacylglutathione hydrolase that catalyses the hydrolysis of S-d-lactoylglutathione to form d-lactic acid and glutathione, which is released from the active site. The tripeptide glutathione is the major sulfhydryl antioxidant and has been shown to control several functions, including S-glutathionylation of proteins. S-Glutathionylation is a way for the cells to store reduced glutathione during oxidative stress, or to protect protein thiol groups from irreversible oxidation, and few enzymes involved in protein S-glutathionylation have been found to date. In this work, the enzyme glyoxalase II and its substrate S-d-lactoylglutathione were incubated with malate dehydrogenase or with actin, resulting in a glutathionylation reaction. Glyoxalase II was also submitted to docking studies. Computational data presented a high propensity of the enzyme to interact with malate dehydrogenase or actin through its catalytic site and further in silico investigation showed a high folding stability of glyoxalase II toward its own reaction product glutathione both protonated and unprotonated. This study suggests that glyoxalase II, through a specific interaction of its catalytic site with target proteins, could be able to perform a rapid and specific protein S-glutathionylation using its natural substrate S-d-lactoylglutathione. SIGNIFICANCE This article reports for the first time a possible additional role of Glo2 that, after interacting with a target protein, is able to promote S-glutathionylation using its natural substrate SLG, a glutathione derived compound. In this perspective, Glo2 can play a new important regulatory role inS-glutathionylation, acquiring further significance in cellular post-translational modifications of proteins.
TL;DR: Results indicated that miR‐24‐3p plays an important role in the initiation and progression of HCC by targeting metallothionein 1M, and the miR-24‐ 3p/metallothionin 1m pathway may contribute to the development of novel therapeutic strategies for HCC in the future.
Abstract: Dysregulation of microRNAs has been demonstrated to contribute to malignant progression of cancers, including hepatocellular carcinoma (HCC). MiR-24-3p was previously reported to be significantly upregulated in HCC. However, the potential role and mechanism of action of miR-24-3p in the initiation and progression of HCC remain largely unknown. Quantitative reverse transcription polymerase chain reaction demonstrated that miR-24-3p was significantly upregulated in HCC tumor tissues compared with nontumor tissues. The cell viability, colony formation assay, and tumorigenicity assays in nude mice showed that miR-24-3p could enhance HCC cell growth in vitro and in vivo. Metallothionein 1M was verified as an miR-24-3p target gene by using dual-luciferase reporter assays, quantitative reverse transcription polymerase chain reaction, and Western blotting, which was involved in miR-24-3p regulated HCC cell growth. These results indicated that miR-24-3p plays an important role in the initiation and progression of HCC by targeting metallothionein 1M, and the miR-24-3p/metallothionein 1M pathway may contribute to the development of novel therapeutic strategies for HCC in the future.
TL;DR: This study is the first to reveal differential lncRNA expression in Bronchopulmonary dysplasia, which provides a foundation for further understanding of the molecular mechanism of BPD development.
Abstract: Bronchopulmonary dysplasia (BPD) is a common complication of premature birth that seriously affects the survival rate and quality of life among preterm neonates. Long non-coding RNAs (lncRNAs) have been implicated in many human diseases. However, the role of lncRNAs in the pathogenesis of BPD remains poorly understood. Here, we exposed neonatal C57BL/6J mice to 95% concentrations of ambient oxygen and established a mouse lung injury model that mimicked human BPD. Next, we compared lncRNA and messenger RNA (mRNA) expression profiles between BPD and normal lung tissues using a high-throughput mouse lncRNA + mRNA array system. Compared with the control group, 882 lncRNAs were upregulated, and 887 lncRNAs were downregulated in BPD lung tissues. We validated some candidate BPD-associated lncRNAs by real-time quantitative reverse-transcription polymerase chain reaction analysis in eight pairs of BPD and normal lung tissues. Gene ontology, pathway and bioinformatics analyses revealed that a downregulated lncRNA, namely AK033210, associated with tenascin C may be involved in the pathogenesis of BPD. To the best of our knowledge, our study is the first to reveal differential lncRNA expression in BPD, which provides a foundation for further understanding of the molecular mechanism of BPD development. Copyright © 2016 John Wiley & Sons, Ltd.
TL;DR: Major evidence is suggested that adipose tissues originating from different anatomic regions are distinguished at the level of the undifferentiated stem cells such as mature adipocytes, suggesting a possibility of ASCs as a therapeutic target against metabolic disorders or cancer.
Abstract: Subcutaneous and visceral adipose tissues show a different risk effect on metabolic disorders because they have distinct cellular properties. We isolated stem cells from the separate human adipose tissues to investigate that subcutaneous and visceral fat depots have metabolic differences. Adipose-derived stem cells (ASCs) were characterized by immunophenotype and differentiation potentials into adipogenic, osteogenic, and chondrogenic lineages. Although subcutaneous and visceral ASCs (S-ASC and V-ASC) express same surface markers (CD31- , CD34- , CD45- , CD73+ , CD90+ , and CD105+ ) and have differentiation potentials, S-ASCs had higher capacity to proliferate and to differentiate into adipogenic lineage than V-ASCs. Next, we identified that S-ASC and V-ASC were genetically distinct based on microarray analysis. Among a total of 810 genes detected in ASCs of both depots, the differentially expressed genes were involved in energy and lipid metabolism. These data show the existence of the intrinsic difference between S-ASC and V-ASC and suggest the differences of anatomically separated adipose tissue. On the basis of the differentially expressed gene profiles between S-ASC and V-ASC, we suggested significant evidence that adipose tissues originating from different anatomic regions are distinguished at the level of the undifferentiated stem cells such as mature adipocytes. V-ASCs had the upregulated clusters of genes related to lipid biosynthesis and metabolism. By contrast, S-ASCs highly expressed genes involved in DNA-dependent transcription, contributing to proliferation. We provide further insights for ASCs with the different origins to understand fat accumulation and distribution and a possibility of ASCs as a therapeutic target against metabolic disorders or cancer.
TL;DR: It is proposed that the TMEM63 proteins may serve as an osmolarity sensitive ion channel for the osmoreception, and all the three members of TMEM 63 proteins are required to constitute a hyperosmolarity activated ion channel.
Abstract: Osmoreception is essential for systemic osmoregulation, a process to stabilize the tonicity and volume of the extracellular fluid through regulating the ingestive behaviour, sympathetic outflow and renal function. The sensation of osmotic changes by osmoreceptor neurons is mediated by ion channels that detect the change of osmolarity in extracellular fluid. However, the molecular identity of these channels remains mysterious. AtCSC1and OSCA1,two closely related paralogues from Arabidopsis, have been demonstrated to form hyperosmolarity activated ion channels, which makes their mammalian orthologues-the members of TMEM63 proteins, possible candidates for osmoreceptor transduction channel. To test this possibility, we cloned the cDNAs of all the three members of the mouse TMEM63 family, TMEM63A, TMEM63B and TMEM63C from the mRNA from mouse brain. When all of the three subtypes of TMEM63 proteins were co-expressed in HEK293 cells, we recorded membrane currents evoked by hypertonic stimulation in these cells. However, the cells expressing the combinations of any two subtypes of TMEM63 proteins could not exhibit any hyperosmolarity evoked currents. Thus, all the three members of TMEM63 proteins are required to constitute a hyperosmolarity activated ion channel. We propose that the TMEM63 proteins may serve as an osmolarity sensitive ion channel for the osmoreception. Copyright © 2016 John Wiley & Sons, Ltd.
TL;DR: Investigation of whether receptor tyrosine kinase‐like orphan receptor 1 (ROR1) is involved in stem cell maintenance and malignant properties in human glioblastoma indicates that targeting ROR1 can induce differentiation of CSCs and inhibit metastasis in gliOBlastoma.
Abstract: Glioblastoma is the most malignant of brain tumours and is difficult to cure because of interruption of drug delivery by the blood-brain barrier system, its high metastatic capacity and the existence of cancer stem cells (CSCs). Although CSCs are present as a small population in malignant tumours, CSCs have been studied as they are responsible for causing recurrence, metastasis and resistance to chemotherapy and radiotherapy for cancer. CSCs have self-renewal characteristics like normal stem cells. The aim of this study was to investigate whether receptor tyrosine kinase-like orphan receptor 1 (ROR1) is involved in stem cell maintenance and malignant properties in human glioblastoma. Knockdown of ROR1 caused reduction of stemness and sphere formation capacity. Moreover, down-regulation of ROR1 suppressed the expression of epithelial-mesenchymal transition-related genes and the tumour migratory and invasive abilities. The results of this study indicate that targeting ROR1 can induce differentiation of CSCs and inhibit metastasis in glioblastoma. In addition, ROR1 may be used as a potential marker for glioblastoma stem cells as well as a potential target for glioblastoma stem cell therapy.
TL;DR: MicroRNA‐383 is a tumor suppressor in human lung cancer by inhibiting EPAS1 and both of which could serve as potential therapeutic targets in the treatment of lung cancer.
Abstract: Lung cancer is the deadliest of all human cancers worldwide. The role of microRNA (miR)-383 has been controversial in the initiation and progression of different cancers. We aimed to investigate the function of miR-383 in human lung cancer both in vitro and in vivo. MicroRNA-383 levels were analyzed in noncancerous versus cancerous human lung tissues and in normal versus lung cancer cell lines. Effect of miR-383 on cell migration and invasion was examined in vitro and on tumor growth by using a xenograft mouse model in vivo. Potential mRNA target of miR-383 was predicted, and underlying mechanism was explored as well. MicroRNA-383 was downregulated in lung cancer tissue and cell lines. Expression of miR-383 inhibited migration and invasion of human lung cancer cell lines in vitro and tumorigenesis of lung cancer xenografts in vivo. MicroRNA-383 directly targeted 3′ untranslated region of endothelial PAS domain-containing protein 1 (EPAS1) messenger RNA and inhibited both its mRNA and protein expressions. Reintroduction of EPAS1 could bypass the inhibition by miR-383 on tumorigenesis of human lung cancer cell lines. MicroRNA-383 is a tumor suppressor in human lung cancer by inhibiting EPAS1, both of which could serve as potential therapeutic targets in the treatment of lung cancer.
Significance of the study
MicroRNA-383 is a tumor suppressor in human lung cancer, which functions to inhibit tumorigenesis of lung cancer cells both in vitro and in vivo. This inhibitory effect is mediated by direct targeting of EPAS1 mRNA and subsequent repressing of its expression. Both microRNA-383 and EPAS1 could serve as potential therapeutic targets in the treatment of lung cancer.
TL;DR: It was found that sevoflurane led to the long‐term cognitive impairment in the young rats and it was shown that α‐lipoic acid offset the effect of sev ofluranes on the neuronal apoptosis and cognitive dysfunction.
Abstract: Sevoflurane is a widely used anaesthetic agent, including in anaesthesia of children and infants. Recent studies indicated that the general anaesthesia might cause the cell apoptosis in the brain. This issue raises the concerns about the neuronal toxicity induced by the application of anaesthetic agents, especially in the infants and young children. In this study, we used Morris water maze, western blotting and immunohistochemistry to elucidate the role of α-lipoic acid in the inhibition of neuronal apoptosis. We found that sevoflurane led to the long-term cognitive impairment in the young rats. This adverse effect may be caused by the neuronal death in the hippocampal region, mediated through PI3K/Akt signalling pathway. We also showed that α-lipoic acid offset the effect of sevoflurane on the neuronal apoptosis and cognitive dysfunction. This study elucidated the potential clinical role of α-lipoic acid, providing a promising way in the prevention and treatment of long-term cognitive impairment induced by sevoflurane general anesthesia.
TL;DR: Insulin‐resistant, but not insulin‐sensitive obese individuals, have an increased percentage of CD16+ monocytes that can be slightly modulated by a single bout of moderate aerobic exercise, which may partially account for the observed reduction in the percentage ofCD16+ cells in response to exercise.
Abstract: UNLABELLED Obesity is a low-grade chronic inflammation condition, and macrophages, and possibly monocytes, are involved in the pathological outcomes of obesity. Physical exercise is a low-cost strategy to prevent and treat obesity, probably because of its anti-inflammatory action. We evaluated the percentage of CD16(-) and CD16(+) monocyte subsets in obese insulin-resistant individuals and the effect of an exercise bout on the percentage of these cells. Twenty-seven volunteers were divided into three experimental groups: lean insulin sensitive, obese insulin sensitive and obese insulin resistant. Venous blood samples collected before and 1 h after an aerobic exercise session on a cycle ergometer were used for determination of monocyte subsets by flow cytometry. Insulin-resistant obese individuals have a higher percentage of CD16(+) monocytes (14.8 ± 2.4%) than the lean group (10.0 ± 1.3%). A positive correlation of the percentage of CD16(+) monocytes with body mass index and fasting plasma insulin levels was found. One bout of moderate exercise reduced the percentage of CD16(+) monocytes by 10% in all the groups evaluated. Also, the absolute monocyte count, as well as all other leukocyte populations, in lean and obese individuals, increased after exercise. This fact may partially account for the observed reduction in the percentage of CD16(+) cells in response to exercise. Insulin-resistant, but not insulin-sensitive obese individuals, have an increased percentage of CD16(+) monocytes that can be slightly modulated by a single bout of moderate aerobic exercise. These findings may be clinically relevant to the population studied, considering the involvement of CD16(+) monocytes in the pathophysiology of obesity. Copyright © 2016 John Wiley & Sons, Ltd. SIGNIFICANCE OF THE STUDY Obesity is now considered to be an inflammatory condition associated with many pathological consequences, including insulin resistance. It is proposed that insulin resistance contributes to the aggravation of the inflammatory dysfunction in obesity. The effect of obesity on the percentage of monocytes was previously observed in class II and III obese individuals who presented other alterations in addition to insulin resistance. In this study we observed that insulin-resistant obese individuals, but not insulin-sensitive ones, had an increased percentage of CD14(+) CD16(+) monocytes. This fact shows that a dysfunction of the monocyte percentage in class I obese individuals is only seen when this condition is associated with insulin resistance.
TL;DR: It is proposed that some replicative senescent cells may avoid apoptosis signalling via impairment of signalling molecules and accumulation during long‐term expansion through resistance to oxidative stress.
Abstract: Cells undergo replicative senescence during in vitro expansion, which is induced by the accumulation of cellular damage caused by excessive reactive oxygen species. In this study, we investigated whether long-term-cultured human bone marrow mesenchymal stromal cells (MSCs) are insensitive to apoptotic stimulation. To examine this, we established replicative senescent cells from long-term cultures of human bone marrow MSCs. Senescent cells were identified based on declining population doublings, increased expression of senescence markers p16 and p53 and increased senescence-associated β-gal activity. In cell viability assays, replicative senescent MSCs in late passages (i.e. 15-19 passages) resisted damage induced by oxidative stress more than those in early passages did (i.e. 7-10 passages). This resistance occurred via caspase-9 and caspase-3 rather than via caspase-8. The senescent cells are gradually accumulated during long-term expansion. The oxidative stress-sensitive proteins ataxia-telangiectasia mutated and p53 were phosphorylated, and the expression of apoptosis molecules Bax increased, and Bcl-2 decreased in early passage MSCs; however, the expression of the apoptotic molecules did less change in response to apoptotic stimulation in late-passage MSCs, suggesting that the intrinsic apoptotic signalling pathway was not induced by oxidative stress in long-term-cultured MSCs. Based on these results, we propose that some replicative senescent cells may avoid apoptosis signalling via impairment of signalling molecules and accumulation during long-term expansion. Copyright © 2016 John Wiley & Sons, Ltd.
TL;DR: The results suggest that Akt and JNK pathways are involved in the regulation of BCMA, and a novel BAFF/BCMA signalling pathway in MM may be a new therapeutic target for MM.
Abstract: B-cell maturation antigen (BCMA) is expressed on normal and malignant plasma cells and represents a potential target for therapeutic intervention. In this study, we characterized the mechanism underlying the protein kinase B (Akt) and c-Jun N-terminal kinase (JNK) pathways and BCMA interactions in regulating multiple myeloma (MM) cell survival. It was found that the expression levels of B cell-activating factor (BAFF) and BCMA were increased in MM cells as compared with those in normal controls. The proliferation of U266 cells was induced by recombinant human BAFF (rhBAFF) and could also be decreased by BCMA siRNA. The expression of Bcl-2 protein was up-regulated, and Bax protein was down-regulated after rhBAFF treatment, which could be reversed by BCMA siRNA. Similarly, the protein p-JNK and p-Akt were activated by rhBAFF and could be changed by BCMA siRNA. In addition, the BCMA mRNA and protein expression levels were decreased after treatment with Akt and JNK pathway inhibitors. These results suggest that Akt and JNK pathways are involved in the regulation of BCMA. A novel BAFF/BCMA signalling pathway in MM may be a new therapeutic target for MM.
TL;DR: The findings reveal that EPB41L3 suppresses tumour cell invasion and inhibits MMP2 and MMP9 expression in ESCC cells.
Abstract: EPB41L3 may play a role as a metastasis suppressor by supporting regular arrangements of actin stress fibres and alleviating the increase in cell motility associated with enhanced metastatic potential. Downregulation of epb41l3 has been observed in many cancers, but the role of this gene in esophageal squamous cell carcinoma (ESCC) remains unclear. Our study aimed to determine the effect of epb41l3 on ESCC cell migration and invasion. We investigated epb41l3 protein expression in tumour and non-tumour tissues by immunohistochemical staining. Expression in the non-neoplastic human esophageal cell line Het-1a and four ESCC cell lines - Kyse150, Kyse510, Kyse450 and Caes17 - was assessed by quantitative Polymerase Chain Reaction (qPCR) and Western blotting. Furthermore, an EPB41L3 overexpression plasmid and EPB41L3-specific small interfering RNA were used to upregulate EPB41L3 expression in Kyse150 cells and to downregulate EPB41L3 expression in Kyse450 cells, respectively. Cell migration and invasion were evaluated by wound healing and transwell assays, respectively. The expression levels of p-AKT, matrix metalloproteinase (MMP)2 and MMP9 were evaluated. Expression of epb41l3 was significantly lower in tumour tissues than in non-tumour tissues and in ESCC cell lines compared with the Het-1a cell line. Kyse450 and Caes17 cells exhibited higher expression of epb41l3 than Kyse150 and Kyse510 cells. Overexpressing epb41l3 decreased Kyse150 cell migration and invasion, whereas EPB41L3-specific small interfering RNA silencing increased these functions in Kyse450 cells. Furthermore, overexpressing epb41l3 led to downregulation of MMP2 and MMP9 in Kyse150 and Kyse510 cells. Our findings reveal that EPB41L3 suppresses tumour cell invasion and inhibits MMP2 and MMP9 expression in ESCC cells.
TL;DR: Results show that the present assay is sensitive and convenient to appreciate ALS and to develop useful agents for the modulation of SOD1 conformation.
Abstract: In the present study, we applied a highly sensitive NanoLuc-based technology to understand the status of superoxide dismutase 1 (SOD1) within mammalian cells. Two fragments of NanoLuc (NanoBit), large N-terminal and small C-terminal regions, were fused with wild-type (wt) and mutant human SOD1 (hSOD1) genes and transfected into cells. Luciferase activity through NanoBit assembly was only detected in NanoBit-tagged wtSOD1-expressing cells. Furthermore, the developed NanoLuc system was used to investigate the role of protein-protein interactions in the pathogenesis of amyotrophic lateral sclerosis (ALS). In addition to SOD1, we also applied this NanoBit system for detecting the dimerization of wild-type, M337V-mutated human TAR-binding protein 43 kDa (hTDP43) and its cleaved C-terminal fragment (TDP25M337V ) as well as their interactions with SOD1. Luciferase activities of NanoBit-tagged mutant SOD1, TDP43, or TDP25 were negligible. Finally, we found that a zinc chelator partially reduced the luciferase activity of NanoBit-wtSOD1. Collectively, these results show that the present assay is sensitive and convenient to appreciate ALS and to develop useful agents for the modulation of SOD1 conformation.
TL;DR: It is suggested that NO‐mediated vasorelaxation plays partly a role in the anabolic action of PTH on cortical bone, as well as the mechanical strength in femurs induced by iPTH.
Abstract: There is strong evidence that vasodilatory nitric oxide (NO) donors have anabolic effects on bone in humans. Parathyroid hormone (PTH), the only osteoanabolic drug currently approved, is also a vasodilator. We investigated whether the NO synthase inhibitor L-NAME might alter the effect of PTH on bone by blocking its vasodilatory effect. BALB/c mice received 28 daily injections of PTH[1-34] (80 µg/kg/day) or L-NAME (30 mg/kg/day), alone or in combination. Hindlimb blood perfusion was measured by laser Doppler imaging. Bone architecture, turnover and mechanical properties in the femur were analysed respectively by micro-CT, histomorphometry and three-point bending. PTH increased hindlimb blood flow by >30% within 10 min of injection (P < 0.001). Co-treatment with L-NAME blocked the action of PTH on blood flow, whereas L-NAME alone had no effect. PTH treatment increased femoral cortical bone volume and formation rate by 20% and 110%, respectively (P < 0.001). PTH had no effect on trabecular bone volume in the femoral metaphysis although trabecular thickness and number were increased and decreased by 25%, respectively. Co-treatment with L-NAME restricted the PTH-stimulated increase in cortical bone formation but had no clear-cut effects in trabecular bone. Co-treatment with L-NAME did not affect the mechanical strength in femurs induced by iPTH. These results suggest that NO-mediated vasorelaxation plays partly a role in the anabolic action of PTH on cortical bone.