Lateral interactions between CD276 and CD147 are essential for stemness in breast cancer: a novel insight from proximal proteome analysis.

Oncogenic cell-surface membrane proteins contribute to the phenotypic and functional characteristics of cancer stem cells (CSCs). We employed a proximity-labeling proteomic approach to quantitatively analyze the cell-surface membrane proteins in close proximity to CD147 in CSCs. Furthermore, we compared CSCs to non-CSCs to identify CSC-specific cell-surface membrane proteins that are closely interact with CD147 and revealed that lateral interaction between CD147 and CD276 concealed within the lipid raft microdomain in CSCs, confers resistance to docetaxel, a commonly used chemotherapy agent for various cancer types, including metastatic breast cancer. Moreover, we investigated the clinical relevance of CD147 and CD276 co-expression in HER2+ breast cancer (BC) and triple-negative breast cancer patients who underwent chemotherapy. We observed poor disease-free survival and Overall survival rates in patients of CD147 and CD276 (p = 0.04 and 0.08, respectively). Subsequent immunohistochemical analysis in independent cohorts of HER2+ BC support for the association between co-expression of CD147 and CD276 and a poor response to chemotherapy. Collectively, our study suggests that the lateral interaction between CD147 and its proximal partners, such as CD276, may serve as a poor prognostic factor in BC and a predictive marker for the critical phenotypic determinant of BC stemness.

RPL17 Promotes Colorectal Cancer Proliferation and Stemness through ERK and NEK2/ß-catenin Signaling Pathways.

Aims: Ribosomal protein L17 (RPL17), a 60S subunit component, is up-regulated in colorectal cancer (CRC). However, its oncogenic role in CRC progression remains unexplored. Thus, we aimed to investigate the effect of RPL17 targeting on CRC in vitro and in vivo and whether RPL17 gained an extra-ribosomal function during CRC development. Methods: RPL17-specific siRNAs complexed with cationic lipids were transfected to CRC cells to silence target gene expression and then real-time RT-PCR and western blotting were applied to observe the change of expression or activity of genes or proteins of interest. Cell proliferation assay, clonogenic assay and cell cycle analysis were used to determine the in vitro effects of RPL17siRNAs on CRC cell growth, and a subcutaneous xenograft assay was applied to test the effect of RPL17siRNAs on in vivo tumor growth. RNA sequencing and western blotting were used to investigate the underlying mechanisms. Sphere-forming assay, invasion assay and migration assay were used to evaluate the effects of RPL17siRNAs on CRC stemness. Results: siRNA-mediated inhibition of RPL17 expression suppressed CRC cell growth and long-term colony formation by inducing apoptotic cell death. Similarly, targeting RPL17 effectively suppressed tumor formation in a mouse xenograft model. RNA sequencing of RPL17-silenced CRC cells revealed the same directional regulation of 159 (93 down- and 66 up-regulated) genes. Notably, NIMA-related kinase 2 (NEK2), which functionally cooperates with extracellular-regulated protein kinase (ERK) and plays a pivotal role in mitotic progression and stemness maintenance, was down-regulated. RPL17 silencing reduced NEK2, ß-catenin, and p-ERK protein levels. These molecular alterations reflected the reduction in sphere-forming capacity, expression of stem cell marker genes, migration, and invasion. Reversely, RPL17 overexpression increased the ability of long-term colony formation, migration, and invasion. Conclusion: Our findings indicate that RPL17 promotes CRC proliferation and stemness via the ERK and NEK2/ß-catenin signaling axis, and targeting RPL17 could be the next molecular strategy for both primary CRC treatment and prevention of secondary tumor formation.

Association of High-Sensitivity C-Reactive Protein and Alcohol Consumption on Metabolic Syndrome in Korean Men.

Background: This study aimed to examine the effect of both alcohol consumption and high-sensitivity C-reactive protein (hsCRP) on metabolic syndrome (MetS) in Korean men. Methods: A cohort of 364 men included in this study was divided into four groups according to the amount of alcohol they consumed: the nondrinkers (ND), low moderate drinkers (LM, ≤7 standard drinks per week), high moderate drinkers (HM, 7 to 14 drinks per week), and heavy drinkers (HD, >14 drinks per week). Logistic regression analyses were performed after adjusting for age, exercise, and smoking. Results: The risk of MetS in the LM group with a high hsCRP level (1.0 or more mg/dL) was not significant. However, the risks of MetS were significantly higher in the HM and HD groups with high hsCRP levels than that in the ND group. The odds ratios of MetS in the HM and HD groups with high hsCRP levels were 3.44 (95% confidence interval (CI), 1.25−9.52) and 3.14 (95% CI, 1.07−9.23), respectively. Conclusion: This study suggests that the risk of MetS is higher in men who consume more than seven drinks a week with high hsCRP levels than that in nondrinkers.

Neisseria gonorrhoeae Multivalent Maxibody with a Broad Spectrum of Strain Specificity and Sensitivity for Gonorrhea Diagnosis.

Gonorrhea is one of the most common, but still hidden and insidious, sexually transmitted diseases caused by Neisseria gonorrhoeae (gonococci). However, the diagnosis and treatment of gonorrhea are hampered by antigenic variability among gonococci, the lack of acquired immunity, and antimicrobial resistance. Further, strains resistant to cephalosporins, including ceftriaxone, the last line of defense, represent a growing threat, which prompted us to develop gonococci-specific diagnostic antibodies with broad-spectrum binding to gonococci strains to generate gonorrhea-detecting reagents. This study reports the identification of gonococci antibodies via bio-panning on gonococci cells using scFv-phage libraries. Reformatting the lead scFv-phage Clones 1 and 4 to a multivalent scFv1-Fc-scFv4 maxibody increased the sensitivity by up to 20-fold compared to the single scFv-Fc (maxibody) alone. Moreover, the multivalent maxibody showed broader cross-reactivity with clinical isolates and the ceftriaxone antibiotic-resistant World Health Organization (WHO) reference strain L. In contrast, the selected antibodies in the scFv-phage, maxibody, and multivalent maxibody did not bind to N. sicca, N. meningitides, and N. lactamica, suggesting the clinical and pharmaceutical diagnostic value of these selected antibodies for gonorrheal infections. The present study illustrates the advantages and potential application of multivalent maxibodies to develop rapid and sensitive diagnostic reagents for infectious diseases and cancer.

Silencing CDCA8 Suppresses Hepatocellular Carcinoma Growth and Stemness via Restoration of ATF3 Tumor Suppressor and Inactivation of AKT/ß-Catenin Signaling.

Big data analysis has revealed the upregulation of cell division cycle associated 8 (CDCA8) in human hepatocellular carcinoma (HCC) and its poorer survival outcome. However, the functions of CDCA8 during HCC development remain unknown. Here, we demonstrate in vitro that CDCA8 silencing inhibits HCC cell growth and long-term colony formation and migration through the accumulation of the G2/M phase cell population. Conversely, CDCA8 overexpression increases the ability to undergo long-term colony formation and migration. RNA sequencing and bioinformatic analysis revealed that CDCA8 knockdown led to the same directional regulation in 50 genes (25 down- and 25 upregulated). It was affirmed based on protein levels that CDCA8 silencing downregulates the levels of cyclin B1 and p-cdc2 and explains how it could induce G2/M arrest. The same condition increased the protein levels of tumor-suppressive ATF3 and GADD34 and inactivated AKT/ß-catenin signaling, which plays an important role in cell growth and stemness, reflecting a reduction in sphere-forming capacity. Importantly, it was demonstrated that the extent of CDCA8 expression is much greater in CD133+ cancer stem cells than in CD133- cancer cells, and that CDCA8 knockdown decreases levels of CD133, p-Akt and ß-catenin and increases levels of ATF3 and GADD34 in the CD133+ cancer stem cell (CSC) population. These molecular changes led to the inhibition of cell growth and sphere formation in the CD133+ cell population. Targeting CDCA8 also effectively suppressed tumor growth in a murine xenograft model, showing consistent molecular alterations in tumors injected with CDCA8siRNA. Taken together, these findings indicate that silencing CDCA8 suppresses HCC growth and stemness via restoring the ATF3 tumor suppressor and inactivating oncogenic AKT/ß-catenin signaling, and that targeting CDCA8 may be the next molecular strategy for both primary HCC treatment and the prevention of metastasis or recurrence.

TNF-α differentially modulates subunit levels of respiratory electron transport complexes of ER/PR +ve/-ve breast cancer cells to regulate mitochondrial complex activity and tumorigenic potential.

BACKGROUND: Tumor necrosis factor-α (TNF-α) is an immunostimulatory cytokine that is consistently high in the breast tumor microenvironment (TME); however, its differential role in mitochondrial functions and cell survival in ER/PR +ve and ER/PR -ve breast cancer cells is not well understood. METHODS: In the current study, we investigated TNF-α modulated mitochondrial proteome using high-resolution mass spectrometry and identified the differentially expressed proteins in two different breast cancer cell lines, ER/PR positive cell line; luminal, MCF-7 and ER/PR negative cell line; basal-like, MDA-MB-231 and explored its implication in regulating the tumorigenic potential of breast cancer cells. We also compared the activity of mitochondrial complexes, ATP, and ROS levels between MCF-7 and MDA-MB-231 in the presence of TNF-α. We used Tumor Immune Estimation Resource (TIMER) webserver to analyze the correlation between TNF-α and mitochondrial proteins in basal and luminal breast cancer patients. Kaplan-Meier method was used to analyze the correlation between mitochondrial protein expression and survival of breast cancer patients. RESULTS: The proteome analysis revealed that TNF-α differentially altered the level of critical proteins of mitochondrial respiratory chain complexes both in MCF-7 and MDA-MB-231, which correlated with differential assembly and activity of mitochondrial ETC complexes. The inhibition of the glycolytic pathway in the presence of TNF-α showed that glycolysis is indispensable for the proliferation and clonogenic ability of MDA-MB-231 cells (ER/PR -ve) as compared to MCF-7 cells (ER/PR +ve). The TIMER database showed a negative correlation between the expressions of TNF-α and key regulators of mitochondrial OXPHOS complexes in basal breast vs lobular carcinoma. Conversely, patient survival analysis showed an improved relapse-free survival with increased expression of identified proteins of ETC complexes and survival of the breast cancer patients. CONCLUSION: The evidence presented in our study convincingly demonstrates that TNF-α regulates the survival and proliferation of aggressive tumor cells by modulating the levels of critical assembly factors and subunits involved in mitochondrial respiratory chain supercomplexes organization and function. This favors the rewiring of mitochondrial metabolism towards anaplerosis to support the survival and proliferation of breast cancer cells. Collectively, the results strongly suggest that TNF-α differentially regulates metabolic adaptation in ER/PR +ve (MCF-7) and ER/PR -ve (MDA-MB-231) cells by modulating the mitochondrial supercomplex assembly and activity.

Targeting CALM2 Inhibits Hepatocellular Carcinoma Growth and Metastasis by Suppressing E2F5-mediated Cell Cycle Progression.

BACKGROUND/AIM: The aim of this study was to reveal the novel roles of calmodulin 2 (CALM2) in hepatocellular carcinoma (HCC) progression. MATERIALS AND METHODS: The effects of knockdown of CALM2 expression by siRNA were investigated using various experimental approaches in both cellular and molecular levels. RESULTS: Silencing of CALM2 inhibited HCC cell proliferation and colony formation through induction of apoptosis. At the molecular level, CALM2-specific knockdown led to the common dysregulation of 154 genes in HCC cells. Notably, E2F transcription factor 5 (E2F5), which is functionally associated with migration, invasion and proliferation, was generally down-regulated. These functional associations were confirmed in HCC clinical samples. Reflecting the molecular changes, CALM2 knockdown reduced the migration and invasion abilities of HCC cells and abrogated the potency of tumor formation in vivo. CONCLUSION: Targeting CALM2 may be a molecular strategy for both primary HCC treatment and prevention of metastasis or recurrence.

Association between Alcohol Consumption and Metabolic Syndrome Determined by Facial Flushing in Korean Women.

BACKGROUND: This study aimed to examine the relationship between alcohol consumption and metabolic syndrome in Korean women as determined by facial flushing. METHODS: Of the female patients aged <65 years who visited the health promotion center at Chungnam National University Hospital from January 2016 to March 2017, 1,344 women were included. After adjusting for confounding factors such as age, body mass index, smoking, exercise, and menopausal status, multiple logistic regression analysis was performed to assess the association between alcohol consumption and the risk of metabolic syndrome in the facial flushing and non-facial flushing groups compared with the non-drinkers. RESULTS: Even after adjusting for confounding factors, the risk of metabolic syndrome was significantly high in all drinking subgroups (≤2 standard drinks: odds ratio [OR], 1.68; 95% confidence interval [CI], 1.10-2.68; 2<, ≤4 standard drinks: OR, 2.48; 95% CI, 1.29-4.74; and >4 standard drinks: OR, 4.16; 95% CI, 2.03-8.30) of the facial flushing group. The risk of metabolic syndrome was significantly high only in the subgroup of weekly alcohol consumption >4 standard drinks (OR, 2.20; 95% CI, 1.07-4.52) in the non-facial flushing group. CONCLUSION: This study suggests that Korean women experiencing facial flushing when drinking have a higher risk of developing metabolic syndrome even with a low weekly drinking amount than those who do not experience face flushing.

Clinicopathological Characteristics of TZAP Expression in Colorectal Cancers.

PURPOSE: The zinc finger protein, ZBTB48, is a telomere-associated protein. It was renamed as telomeric zinc finger-associated protein (TZAP) binding to elongated telomeres. However, its expression level was not measured in cancers. PATIENTS AND METHODS: We analyzed TZAP mRNA levels in 60 colorectal cancers (CRC) and its correlation with telomere length and TERT was studied. RESULTS: TZAP mRNA in CRC was higher statistically than that in paired non-cancerous tissues (p = 0.033). Higher TZAP was found in carcinoembryonic antigen (CEA)-positive CRCs (>5 ng/mL) (p = 0.012). Shorter telomere was found in CRCs with high TZAP expression than that with low TZAP expression (p = 0.010). According to quantitative correlation analysis, TZAP has a correlation with age (r = -0.349, p = 0.007), TERT (r = 0.279, p = 0.041) and telomere length (r = -0.305, p = 0.021). TZAP expression did not harbor prognostic value in CRC. Inhibition of TZAP expression by siRNA suppresses cell growth in HT29 cells; however, it resulted in increased cell viability in HCT116 cells. TZAP inhibition induces a decrease in mRNA levels of TERT in both HT29 and HCT116 cells. TCGA data analysis showed higher expression of TZAP showed poorer overall survival in colon cancer (p = 0.001); however, it did not have a significance in rectal cancer (p = 0.951). CONCLUSION: We suggested that TZAP may be a possible biomarker for CRC.

Human adipose-derived mesenchymal stem cell-based medical microrobot system for knee cartilage regeneration in vivo.

Targeted cell delivery by a magnetically actuated microrobot with a porous structure is a promising technique to enhance the low targeting efficiency of mesenchymal stem cell (MSC) in tissue regeneration. However, the relevant research performed to date is only in its proof-of-concept stage. To use the microrobot in a clinical stage, biocompatibility and biodegradation materials should be considered in the microrobot, and its efficacy needs to be verified using an in vivo model. In this study, we propose a human adipose-derived MSC-based medical microrobot system for knee cartilage regeneration and present an in vivo trial to verify the efficacy of the microrobot using the cartilage defect model. The microrobot system consists of a microrobot body capable of supporting MSCs, an electromagnetic actuation system for three-dimensional targeting of the microrobot, and a magnet for fixation of the microrobot to the damaged cartilage. Each component was designed and fabricated considering the accessibility of the patient and medical staff, as well as clinical safety. The efficacy of the microrobot system was then assessed in the cartilage defect model of rabbit knee with the aim to obtain clinical trial approval.

Influence of Facial Flushing on Pre- or Type 2 Diabetes Risk according to Alcohol Consumption in Korean Male.

BACKGROUND: This study aims to examine the association between alcohol consumption and the risk of pre- or type 2 diabetes mellitus (T2DM) by alcohol-induced flushing response in Korean male adults, particularly based on their body mass index (BMI). METHODS: This study selected 1,030 (158 non-drinkers, 364 flushers, and 508 non-flushers) male adults who had medical checkups. A logistic regression analysis was used to compare the association between alcohol consumption and the risk of pre- or T2DM. RESULTS: In both the normal-weight group (BMI <23 kg/m2) and the overweight group (BMI ≥23 kg/m2 and <25 kg/ m2), the flushers had a higher risk of pre- or T2DM (odds ratio, 95% confidence interval) when consuming more than 8 drinks of alcohol per week than the non-drinkers (normal-weight group: 3.43, 1.06-11.07; overweight group: 4.94, 1.56-15.67). But in the non-flushers among the normal-weight group and the overweight group, there was no significant difference compared to non-drinkers regarding the risk of pre- or T2DM. Obese flushers had a significantly higher risk of pre- or T2DM when consuming more than 4 drinks of alcohol per week than the non-drinkers (>4 and ≤8 drinks: 2.64, 1.10-6.36; >8 drinks: 2.42, 1.11-5.27). However, obese non-flushers had only a significant higher risk of pre- or T2DM when consuming more than 8 drinks of alcohol per week than the non-drinkers (2.72, 1.39-5.30). CONCLUSION: These results suggest that obese flushers have an increased risk of developing pre- or T2DM even with less alcohol consumption.

Influence of Maitake (Grifola frondosa) Particle Sizes on Human Mesenchymal Stem Cells and In Vivo Evaluation of Their Therapeutic Potential.

Maitake (Grifola frondosa) mushroom has received an enormous amount of attention as a dietary supplement due to its high nutritional values. The particle sizes of G. frondosa mushrooms were monitored by a classifying mill. ß-Glucans are the bioactive component of the mushroom, and it was revealed through Fourier transform infrared spectroscopy (FTIR), proton and carbon nuclear magnetic resonance (1H and 13C-NMR), matrix-assisted laser desorption/ionization, and time-of-flight (MALDI-TOF) spectrometry. The biocompatibility of G. frondosa particles, as well as induced osteogenesis of hMSCs, was evaluated through WST-1 assay and alizarin staining (ARS) technique, respectively. Notably, enhanced cell viability was noted in the presence of G. frondosa. Significantly improved calcium deposition has observed from hMSCs with G. frondosa, suggesting to their mineralization potential. The expression of osteogenic related gene markers was examined in the presence of G. frondosa through real-time polymerase chain reaction (qPCR) technique. The upregulation of osteogenic gene markers in the presence of G. frondosa particles was indicating their superior osteogenic potential. Besides, G. frondosa also activated the secretion of various kinds of proteins from the hMSCs indicating their potential for tissue engineering applications. Enhanced secretion of different immunoglobulins was observed in rat serum in the presence of G. frondosa, further demonstrating their therapeutic nature. Therefore, G. frondosa is effective for enhanced osteogenesis and can be utilized as a natural, edible, and osteogenic agent.

O-Linked N-Acetylglucosamine Modification of Mitochondrial Antiviral Signaling Protein Regulates Antiviral Signaling by Modulating Its Activity.

Post-translational modifications, including O-GlcNAcylation, play fundamental roles in modulating cellular events, including transcription, signal transduction, and immune signaling. Several molecular targets of O-GlcNAcylation associated with pathogen-induced innate immune responses have been identified; however, the direct regulatory mechanisms linking O-GlcNAcylation with antiviral RIG-I-like receptor signaling are not fully understood. In this study, we found that cellular levels of O-GlcNAcylation decline in response to infection with Sendai virus. We identified a heavily O-GlcNAcylated serine-rich region between amino acids 249-257 of the mitochondrial antiviral signaling protein (MAVS); modification at this site disrupts MAVS aggregation and prevents MAVS-mediated activation and signaling. O-GlcNAcylation of the serine-rich region of MAVS also suppresses its interaction with TRAF3; this prevents IRF3 activation and production of interferon-ß. Taken together, these results suggest that O-GlcNAcylation of MAVS may be a master regulatory event that promotes host defense against RNA viruses.

The Effect of Education and Vitamin D Supplementation on the Achievement of Optimal Vitamin D Level in Korean Postmenopausal Women.

BACKGROUND: Despite daily vitamin D recommendations, women with osteoporosis may not achieve optimal 25-hydroxy-vitamin D (25[OH]D) levels. We retrospectively evaluated the effect of education and vitamin D supplementation (1,000 IU/day) in Korean women with osteoporosis. METHODS: Sixty-one women with osteoporosis who were taking cholecalciferol (800-1,000 IU/day) were enrolled during 2011 to 2012. Forty patients (education only, Edu group) were educated on the importance of >30 min sunlight exposure daily while taking vitamin D. Twenty-one patients (education with vitamin D supplementation, Add group) were prescribed 1,000 IU/day cholecalciferol (total 1,800-2,000 IU/day) plus education. Patients were divided into 3 groups according to serum 25(OH)D status: deficiency (<20 ng/mL), insufficiency (20-30 ng/mL), and sufficiency (≥30 ng/mL). Furthermore, 25(OH)D levels were compared at baseline and after intervention for 3 months. RESULTS: The median (interquartile range) serum 25(OH)D concentration at baseline was 25.10 (18.95-33.60) ng/mL. The mean (±standard error) differences in 25(OH)D levels from baseline to post-intervention were 19.85±3.86 and 31.73±4.82 ng/mL in the Edu group and Add group, respectively. Eighteen patients (29.5%) had vitamin D deficiency, 25 (41.0%) had insufficiency, and 18 (29.5%) had sufficient levels. Optimal 25(OH)D (30 ng/mL or more) was achieved in 54.5% and 95.2% patients in the Edu group and Add group, respectively (P=0.003). CONCLUSIONS: We consider that vitamin D concentration should be measured on a regular basis in order to maintain an optimal level of vitamin D concentration, and education and supplementation is needed if not sufficient.

Stimuli-Responsive Graphene Nanohybrids for Biomedical Applications.

Stimuli-responsive materials, also known as smart materials, can change their structure and, consequently, original behavior in response to external or internal stimuli. This is due to the change in the interactions between the various functional groups. Graphene, which is a single layer of carbon atoms with a hexagonal morphology and has excellent physiochemical properties with a high surface area, is frequently used in materials science for various applications. Numerous surface functionalizations are possible for the graphene structure with different functional groups, which can be used to alter the properties of native materials. Graphene-based hybrids exhibit significant improvements in their native properties. Since functionalized graphene contains several reactive groups, the behavior of such hybrid materials can be easily tuned by changing the external conditions, which is very useful in biomedical applications. Enhanced cell proliferation and differentiation of stem cells was reported on the surfaces of graphene-based hybrids with negligible cytotoxicity. In addition, pH or light-induced drug delivery with a controlled release rate was observed for such nanohybrids. Besides, notable improvements in antimicrobial activity were observed for nanohybrids, which demonstrated their potential for biomedical applications. This review describes the physiochemical properties of graphene and graphene-based hybrid materials for stimuli-responsive drug delivery, tissue engineering, and antimicrobial applications.

Structural Elucidation and Immune-Enhancing Effects of Novel Polysaccharide from Grifola frondosa.

Beta-glucan (ß-glucan) is a macromolecule structure where glucose unit has bonded through ß-glycosidic bond at 1 and 3 positions. It is well known as a natural immunomodulator without exhibiting any side effects via enhancing immunity. Mushroom contains a large amount of ß-glucan and it has anticancerous and antioxidant efficacy. Structure and physical properties of ß-glucan are highly influenced by the types of mushroom. In particular, Grifola frondosa has ß-1, 3 and ß-1, 6 bonds in their structure. It has been noted that ß-glucan content also depends upon the size of mushroom particles. The exact content of ß-glucan and their immunological activity by a particle size of G. frondosa have yet to be fully elucidated. Herein, ß-glucan contents were analyzed according to the particle size of leaf mushroom followed by cell activation and immunoactivity analysis. The highest ß-glucan content was observed at a particle size of 20-30 µm (27.65 ± 0.30 w/w). All samples showed ~ 103% cell activation compared to the control and greater cell activity was observed at higher concentration. The significant increase in cytokines secretion was observed in the presence of 20-30 µm particle size of G. frondosa compared to the control. This study suggested that 20-30 µm size is the suitable size of G. frondosa that can be used as a health supplement and food additive to act as an immune booster, hypotensive agent, and hypoglycemic agent.

Enhanced osteogenesis of mesenchymal stem cells on electrospun cellulose nanocrystals/poly(ε-caprolactone) nanofibers on graphene oxide substrates.

Cellulose nanocrystals (CNCs) have received a great amount of attention to the production of micro/nano-platforms for tissue engineering applications. CNCs were extracted from rice husk biomass and characterized by different spectroscopic techniques. The biocompatibility of the extracted CNCs was revealed by the WST-1 assay technique in the presence of human mesenchymal stem cells (hMSCs) after different time intervals. An improvement in the mechanical properties was observed in the fabricated scaffolds (PCL/CNC) compared to PCL scaffolds. Graphene oxide (GO)-coated (PCL/CNC) electrospun scaffolds (GPC) were prepared by the deposition of PCL/CNC composite nanofibers on the surface of GO for tissue engineering. Notably, better cell proliferation and differentiation were observed in the presence of the fabricated scaffolds. This enhancement of the properties of the fabricated scaffolds was due to the presence of conductive GO moieties which facilitated the cellular response. Therefore, the fabricated materials have the potential to be used as a biomaterial for enhanced cell proliferation and osteogenic differentiation.

A subset of microRNAs defining the side population of a human malignant mesothelioma cell line.

This study was performed to investigate the global expression profile of microRNAs in distinct subpopulations of a human malignant mesothelioma cell line. Total RNAs were isolated from the sorted side population and non-side population of MS1. The RNAs were subjected to analysis using Affymetrix GeneChip microRNA Arrays. After data extraction and normalization, a subset of microRNAs defining cell subpopulations was identified using bioinformatics softwares. Based on the criteria of 2-fold difference and the p-value of < 0.05, a total of 95 microRNAs were differentially expressed in the side population compared to the non-side population. Functional ontology revealed that target genes of the miRNAs were categorized into various gene ontology terms, such as stem cell maintenance, cell proliferation, programmed cell death, cell migration, and cellular response to stress. The Kyoto Encyclopedia of Genes and Genomes analysis showed that ErbB-2 receptor tyrosine kinases signaling pathway was the most represented. Integrated analysis of MiRTarBase and RNA-seq identified 12 target genes of microRNAs defining side population, including DDIT4 and ROCK2. The present study indicates that a distinct set of microRNAs may be critically involved in the generation and maintenance of heterogeneous subpopulations of cancer cells. They could be a plausible target for the eradication of more aggressive cancer cell subpopulations.

An Integrated Analysis of the Genome-Wide Profiles of DNA Methylation and mRNA Expression Defining the Side Population of a Human Malignant Mesothelioma Cell Line.

Intratumoral heterogeneity is a hallmark of all cancers and functions as the major barrier against effective cancer therapy. In contrast to genetic mutations, the role of epigenetic modifications in the generation and maintenance of heterogeneous cancer cells remains largely undetermined. This study was performed to evaluate the epigenetic mechanisms involved in the tumor cell heterogeneity using side population (SP) and non-SP cells isolated from a human malignant mesothelioma (HMM) cell line. The subpopulations of cancer cells were analyzed by methylated DNA immunoprecipitation combined with high-throughput sequencing (MeDIP-seq) and RNA-seq methodology. The RNA-seq data were analyzed with the MeDIP-seq data in an integrated way to identify the epigenetically modified genes that defined the SP. Concomitant changes in mRNA expression and DNA methylation were found in 122 genes, including 118 down-regulated genes with hypermethylation and 4 up-regulated genes with hypomethylation. Gene ontology revealed that a large portion of the genes belonged to the groups of biological processes such as stem cell maintenance, stem cell development, stem cell differentiation, and the negative regulation of the developmental process. Among these genes, BNC1, RPS6KA3, TWSG1 and DUSP15 contained aberrant methylation in the CpG islands of the promoter region, indicating that the genes regulated by DNA methylation characterized a distinct subpopulation of HMM cells. The present study provided valuable information to shed light on the epigenetic contributions to the generation and maintenance of tumor cell heterogeneity.

Differences in metabolic biomarkers in the blood and gene expression profiles of peripheral blood mononuclear cells among normal weight, mildly obese and moderately obese subjects.

We compared metabolic biomarkers in the blood and peripheral blood mononuclear cell (PBMC) gene expression profiles among normal weight (BMI, 18·5-23 kg/m2), mildly obese (BMI, 25-27·5 kg/m2) and moderately obese Korean adult men (BMI, 27·5-30 kg/m2). High leptin, lipids (except LDL- and HDL-cholesterol) and apoB levels and low adiponectin and HDL-cholesterol levels were present in the plasma of both mildly and moderately obese subjects. Circulating levels of inflammatory cytokines and markers of insulin resistance, oxidative stress and liver damage were altered in moderately obese subjects but not in mildly obese subjects. PBMC transcriptome data showed enrichment of pathways involved in energy metabolism, insulin resistance, bone metabolism, cancer, inflammation and fibrosis in both mildly and moderately obese subjects. Signalling pathways involved in oxidative phosphorylation, TAG synthesis, carbohydrate metabolism and insulin production; mammalian target of rapamycin, forkhead box O, ras-proximate-1, RAS and transforming growth factor-ß signalling; as well as extracellular matrix-receptor interaction were enriched only in moderately obese subjects, indicating that changes in PBMC gene expression profiles, according to metabolic disturbances, were associated with the development and/or aggravation of obesity. In particular, fourteen and fifteen genes differentially expressed only in mildly obese subjects and in both mildly and moderately obese subjects, respectively, could be used as early or stable biomarkers for diagnosing and treating obesity-associated metabolic disturbance. We characterised BMI-associated metabolic and molecular biomarkers in the blood and provided clues about potential blood-based targets for preventing or treating obesity-related complications.