ABSTRACT
The translation elongation factor eEF1A promotes protein synthesis. Its methylation by METTL13 increases its activity, supporting tumor growth. However, in some cancers, a high abundance of eEF1A isoforms is associated with a good prognosis. Here, we found that eEF1A2 exhibited oncogenic or tumor-suppressor functions depending on its interaction with METTL13 or the phosphatase PTEN, respectively. METTL13 and PTEN competed for interaction with eEF1A2 in the same structural domain. PTEN-bound eEF1A2 promoted the ubiquitination and degradation of the mitosis-promoting Aurora kinase A in the S and G2 phases of the cell cycle. eEF1A2 bridged the interactions between the SKP1-CUL1-FBXW7 (SCF) ubiquitin ligase complex, the kinase GSK3ß, and Aurora-A, thereby facilitating the phosphorylation of Aurora-A in a degron site that was recognized by FBXW7. Genetic ablation of Eef1a2 or Pten in mice resulted in a greater abundance of Aurora-A and increased cell cycling in mammary tumors, which was corroborated in breast cancer tissues from patients. Reactivating this pathway using fimepinostat, which relieves inhibitory signaling directed at PTEN and increases FBXW7 expression, combined with inhibiting Aurora-A with alisertib, suppressed breast cancer cell proliferation in culture and tumor growth in vivo. The findings demonstrate a therapeutically exploitable, tumor-suppressive role for eEF1A2 in breast cancer.
Subject(s)
Aurora Kinase A , Breast Neoplasms , Mammary Neoplasms, Animal , PTEN Phosphohydrolase , Peptide Elongation Factor 1 , Animals , Female , Humans , Mice , Aurora Kinase A/genetics , Aurora Kinase A/metabolism , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , F-Box-WD Repeat-Containing Protein 7/genetics , Glycogen Synthase Kinase 3 beta , Mammary Neoplasms, Animal/genetics , Mammary Neoplasms, Animal/metabolism , Mammary Neoplasms, Animal/pathology , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/metabolism , Peptide Elongation Factor 1/genetics , Peptide Elongation Factor 1/metabolismABSTRACT
Microalgae are attracting attention as a next-generation alternative source of protein and essential fatty acids that do not consume large amounts of water or land. Chaetoceros gracilis (C. gracilis)-a marine microalga-is rich in proteins, fucoxanthin, and eicosapentaenoic acid (EPA). Growing evidence indicates that dietary fucoxanthin and EPA have beneficial effects in humans. However, none of these studies have shown that dietary C. gracilis has beneficial effects in mammals. In this study, we investigated the effects of dietary C. gracilis on lipid abnormalities in Sprague-Dawley rats fed a high-sucrose cholesterol-containing diet. Dried C. gracilis was added to the control diet at a final dose of 2 or 5% (w/w). After four weeks, the soleus muscle weights were found to be dose-responsive to C. gracilis and showed a tendency to increase. The hepatic triglyceride and total cholesterol levels were significantly reduced by C. gracilis feeding compared to those in the control group. The activities of FAS and G6PDH, which are related to fatty acid de novo synthesis, were found to be dose-responsive to C. gracilis and tended to decrease. The hepatic glycerol content was also significantly decreased by C. gracilis feeding, and the serum HDL cholesterol levels were significantly increased, whereas the serum levels of cholesterol absorption markers (i.e., campesterol and ß-sitosterol) and the hepatic mRNA levels of Scarb1 were significantly decreased. Water-soluble metabolite analysis showed that the muscular contents of several amino acids, including leucine, were significantly increased by C. gracilis feeding. The tendency toward an increase in the weight of the soleus muscle as a result of C. gracilis feeding may be due to the enhancement of muscle protein synthesis centered on leucine. Collectively, these results show that the oral administration of C. gracilis alleviates hepatic lipid accumulation in rats fed a high-sucrose and cholesterol-containing diet, indicating the potential use of C. gracilis as a food resource.
ABSTRACT
This study was designed to profile the metabolites of Isochrysis galbana, an indigenous and less explored microalgae species. 1H Nuclear Magnetic Resonance (NMR) spectroscopy and Liquid Chromatography-Mass Spectrometry (LCMS) were used to establish the metabolite profiles of five different extracts of this microalga, which are hexane (Hex), ethyl acetate (EtOAc), absolute ethanol (EtOH), EtOH:water 1:1 (AqE), and 100% water (Aq). Partial least square discriminant analysis (PLS-DA) of the generated profiles revealed that EtOAc and Aq extracts contain a diverse range of metabolites as compared to the other extracts with a total of twenty-one metabolites, comprising carotenoids, polyunsaturated fatty acids, and amino acids, that were putatively identified from the NMR spectra. Meanwhile, thirty-two metabolites were successfully annotated from the LCMS/MS data, ten of which (palmitic acid, oleic acid, α-linolenic acid, arachidic acid, cholesterol, DHA, DPA, fucoxanthin, astaxanthin, and pheophytin) were similar to those present in the NMR profile. Another eleven glycerophospholipids were discovered using MS/MS-based molecular network (MN) platform. The results of this study, besides providing a better understanding of I.galbana's chemical make-up, will be of importance in exploring this species potential as a feed ingredient in the aquaculture industry.
Subject(s)
Haptophyta/metabolism , Metabolomics , Amino Acids/isolation & purification , Carotenoids/isolation & purification , Chromatography, Liquid , Fatty Acids, Unsaturated/isolation & purification , Magnetic Resonance Spectroscopy , Mass Spectrometry , Tandem Mass SpectrometryABSTRACT
Microalgae can use either ammonium or nitrate for its growth and vitality. However, at a certain level of concentration, ammonium nitrogen exhibits toxicity which consequently can inhibit microalgae productivity. Therefore, this study is aimed to investigate the tolerance of Tetraselmis tetrathele to high ammonium nitrogen concentrations and its effects on growth rate, photosynthetic efficiency (F v /F m ), pigment contents (chlorophyll a, lutein, neoxanthin, and ß-carotene), and fatty acids production. Experiments were performed at different ammonium nitrogen concentrations (0.31-0.87 gL-1) for 6 days under a light source with an intensity of 300 µmol photons m-2 s-1 and nitrate-nitrogen source as the experimental control. The findings indicated no apparent enhancement of photosynthetic efficiency (F v/F m) at high levels of ammonium nitrogen ( NH 4 + -N) for T. tetrathele within 24 h. However, after 24 h, the photosynthetic efficiency of T. tetrathele increased significantly (p < 0.05) in high concentration of NH 4 + -N. Chlorophyll a content in T. tetrathele grown in all of the different NH 4 + -N levels increased significantly compared to nitrate-nitrogen (NO3-N) treatment (p < 0.05); which supported that this microalgal could grow even in high level of NH 4 + -N concentrations. The findings also indicated that T. tetrathele is highly resistant to high ammonium nitrogen which suggests T. tetrathele to be used in the aquaculture industry for bioremediation purpose to remove ammonium nitrogen, thus reducing the production cost while improving the water quality.
ABSTRACT
BACKGROUND: Cationic amphiphilic chitosan derivatives can form polymeric micelles, which are useful cosmetic materials, but they form polyion complexes with anionic polymers, which can cause formulation difficulties. AIMS: This study aimed to evaluate the usefulness of partially myristoylated carboxymethyl chitosan, an amphoteric-amphiphilic chitosan derivative, as a new material for cosmetics in the absence of a surfactant comprising an anionic polymer. METHODS: An anionic polymer and 1,2-decanediol (an antimicrobial agent)-containing partially myristoylated carboxymethyl chitosan nanoemulsified lotion and glabridin (an antimelanogenic agent)-containing partially myristoylated carboxymethyl chitosan polymeric micelle were prepared using a pressure homogenization method. The release of interleukin-1α, cell viability, and melanogenesis inhibition was evaluated on a human skin model. Antimicrobial activity was evaluated using agar dilution method. RESULTS: A mixture of partially myristoylated carboxymethyl chitosan and carboxyvinyl polymer did not form a polyion complex, but it formed a hydrophilic gel. The anionic polymer-containing partially myristoylated carboxymethyl chitosan nanoemulsified formulation was stable, with no decrease in cell viability and horny layer exfoliation, which are typically observed with Tween 60. Compared with the formulation with methyl paraben (0.2%), the formulation to which 1,2-decanediol (0.05%) was added improved the antibacterial activity against methicillin-resistant Staphylococcus aureus and Propionibacterium acnes; however, no interleukin-1α upregulation was observed. The glabridin-containing partially myristoylated carboxymethyl chitosan polymeric micelles enhanced melanogenesis inhibition and percutaneous glabridin delivery to the epidermis compared with conventional emulsified micelles. CONCLUSIONS: These results suggest that partially myristoylated carboxymethyl chitosan-forming polymeric micelles, in combination with 1,2-decanediol and glabridin, may be useful for surfactant-free cosmetic emulsions.
Subject(s)
Chitosan , Cosmetics , Methicillin-Resistant Staphylococcus aureus , Chitosan/pharmacology , Cosmetics/pharmacology , Drug Carriers , Humans , Micelles , Particle SizeABSTRACT
Significantly high eicosapentaenoic acid (EPA) and fucoxanthin contents with high production rate were achieved in semi continuous culture of marine diatom. Effects of dilution rate on the production of biomass and high value biocompounds such as EPA and fucoxanthin were evaluated in semi-continuous cultures of Chaetoceros gracilis under high light condition. Cellular dry weight increased at lower dilution rate and higher light intensity conditions, and cell size strongly affected EPA and fucoxanthin contents. The smaller microalgae cells showed significantly higher (p < 0.05) value of 17.1 mg g-dw-1 fucoxanthin and 41.5% EPA content per total fatty acid compared to those observed in the larger cells. Chaetoceros gracilis can accumulate relatively higher EPA and fucoxanthin than those reported previously. In addition, maintenance of small cell size by supplying sufficient nutrients and light energy can be the key for the increase production of valuable biocompounds in C. gracilis.
ABSTRACT
Chlorella can produce an unusually wide range of metabolites under various nutrient availability, carbon source, and light availability. Glucose, an essential molecule for the growth of microorganisms, also contributes significantly to the metabolism of various metabolic compounds produced by Chlorella. In addition, manipulation of light intensity also induces the formation of secondary metabolites such as pigments, and carotenoids in Chlorella. This study will focus on the effect of glucose addition, and moderate light on the regulation of carotenoid, lipid, starch, and other key metabolic pathways in Chlorella sorokiniana. To gain knowledge about this, we performed transcriptome profiling on C. sorokiniana strain NIES-2168 in response to moderate light stress supplemented with glucose under mixotrophic conditions. A total of 60,982,352 raw paired-end (PE) reads 100 bp in length was obtained from both normal, and mixotrophic samples of C. sorokiniana. After pre-processing, 93.63% high-quality PE reads were obtained, and 18,310 predicted full-length transcripts were assembled. Differential gene expression showed that a total of 937, and 1124 genes were upregulated, and downregulated in mixotrophic samples, respectively. Transcriptome analysis revealed that the mixotrophic condition caused upregulation of genes involved in carotenoids production (specifically lutein biosynthesis), fatty acid biosynthesis, TAG accumulation, and the majority of the carbon fixation pathways. Conversely, starch biosynthesis, sucrose biosynthesis, and isoprenoid biosynthesis were downregulated. Novel insights into the pathways that link the enhanced production of valuable metabolites (such as carotenoids in C. sorokiniana) grown under mixotrophic conditions is presented.
Subject(s)
Chlorella/genetics , Glucose/metabolism , Light , Transcriptome , Chlorella/metabolism , Genes, Plant , RNA, Plant/geneticsABSTRACT
Chlorella is a popular microalga with robust physiological and biochemical characteristics, which can be cultured under various conditions. The exploration of the small RNA content of Chlorella could improve strategies for the enhancement of metabolite production from this microalga. In this study, stress was introduced to the Chlorella sorokiniana culture to produce high-value metabolites such as carotenoids and phenolic content. The small RNA transcriptome of C. sorokiniana was sequenced, focusing on microRNA (miRNA) content. From the analysis, 98 miRNAs were identified in cultures subjected to normal and stress conditions. The functional analysis result showed that the miRNA targets found were most often involved in the biosynthesis of secondary metabolites, followed by protein metabolism, cell cycle, and porphyrin and chlorophyll metabolism. Furthermore, the biosynthesis of secondary metabolites such as carotenoids, terpenoids, and lipids was found mostly in stress conditions. These results may help to improve our understanding of regulatory mechanisms of miRNA in the biological and metabolic process of Chlorella species. It is important and timely to determine the true potential of this microalga species and to support the potential for genetic engineering of microalgae as they receive increasing focus for their development as an alternative source of biofuel, food, and health supplements.
Subject(s)
Chlorella/genetics , Gene Expression Regulation, Plant , High-Throughput Nucleotide Sequencing/methods , MicroRNAs/genetics , Plant Proteins/genetics , Transcriptome , Chlorella/growth & development , Chlorella/metabolism , Gene Expression Profiling , Plant Proteins/metabolismABSTRACT
Although many metabolomics studies of higher land plant species have been conducted, similar studies of lower nonland plant species, which include microalgae, are still developing. The present study represents an attempt to characterize the metabolic profile of a microalgal diatom Chaetoceros calcitrans, by applying high-resolution mass spectrometry detection, via Q-ExactiveTM Plus Orbitrap mass spectrometry. The results showed that 54 metabolites of various classes were tentatively identified. Experimentally, the chloroform and acetone extracts were clearly distinguished from other solvent extracts in chemometric regression analysis using PLS, showing the differences in the C. calcitrans metabolome between the groups. In addition, specific metabolites were evaluated, which supported the finding of antioxidant and anti-inflammatory activities. This study also provides data on the quantitative analysis of four carotenoids based on the identification results. Therefore, these findings could serve as a reliable tool for identifying and quantifying the metabolome that could reflect the metabolic activities of C. calcitrans.
Subject(s)
Diatoms/metabolism , Metabolome , Metabolomics , Microalgae/metabolism , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry , Animals , Anti-Inflammatory Agents/metabolism , Anti-Inflammatory Agents/pharmacology , Antioxidants/metabolism , Antioxidants/pharmacology , Chromatography, High Pressure Liquid , Macrophages/drug effects , Macrophages/metabolism , Mice , Nitric Oxide/metabolism , RAW 264.7 Cells , Solvents/chemistryABSTRACT
We isolated fifty-two strains from the marine aquaculture ponds in Malaysia that were evaluated for their lipid production and ammonium tolerance and four isolates were selected as new ammonium tolerant microalgae with high-lipid production: TRG10-p102 Oocystis heteromucosa (Chlorophyceae); TRG10-p103 and TRG10-p105 Thalassiosira weissflogii (Bacillariophyceae); and TRG10-p201 Amphora coffeiformis (Bacillariophyceae). Eicosapentenoic acid (EPA) in three diatom strain was between 2.6 and 18.6 % of total fatty acids, which were higher than in O. heteromucosa. Only A. coffeiformi possessed arachidonic acid. Oocystis heteromucosa naturally grew at high ammonium concentrations (1.4-10â¯mM), whereas the growth of the other strains, T. weissflogii and A. coffeiformi, were visibly inhibited at high ammonium concentrations (>1.4â¯mM-NH4). However, two strains of T. weissflogii were able to grow at up to 10â¯mM-NH4 by gradually acclimating to higher ammonium concentrations. The ammonium tolerant strains, especially T. weissflogii which have high EPA contents, were identified as a valuable candidate for biomass production utilizing NH4-N media, such as ammonium-rich wastewater.
Subject(s)
Ammonium Compounds/metabolism , Aquaculture/methods , Bioprospecting/methods , Microalgae/metabolism , Ponds/microbiology , Wastewater/microbiology , Ammonium Compounds/adverse effects , Biodegradation, Environmental , Biofuels/microbiology , Biomass , Cell Culture Techniques/methods , Diatoms/metabolism , Fatty Acids/analysis , Lipids/biosynthesis , Malaysia , Microalgae/drug effects , Microalgae/growth & development , Water Pollutants, Chemical , Water PurificationABSTRACT
The primary biological treatment method for organic sludge is composting and/or anaerobic digestion, but their product (compost or biogas) is of little economic benefit; therefore, an improved process to produce a high-value product is required to make sludge management more sustainable. Maximizing NH3 gas recovery during composting processes has the potential benefit of producing high-value microalgal biomass. However, the majority of produced ammonia does not evaporate as NH3 gas but retains as NH4+-N in the compost after fermentation. The present study investigates the effects of the timing of Ca(OH)2 dosing (on days 2, 5, and 9), and the Ca(OH)2 dose (1.1-2.6â¯mmol/batch), on lab-scale thermophilic composting of anaerobic sludge. The effects on NH3 recovery, organic matter degradability, and microbial activity are evaluated. Ca(OH)2 dosing immediately improved the emission of NH3, with yields 50-69% higher than those under control conditions. The timing of the dosing did not influence NH3 recovery or organic matter degradability. Higher Ca(OH)2 doses resulted in higher NH3 recovery, while microbial activity was temporarily and marginally inhibited. The pH of the compost reached 10-11.5 but quickly dropped to 8-8.5 within a day, probably because of neutralization of Ca(OH)2 by the emitted CO2 and release of NH3, which maintained the microbial activity. The present study indicated that Ca(OH)2 dosing would be useful to apply during thermophilic composting for NH3 recovery to cultivate high-value microalgal biomass, which enables this process to obtain a more economic benefit.
Subject(s)
Ammonia/analysis , Calcium Hydroxide/chemistry , Composting , Sewage/analysis , Anaerobiosis , Dose-Response Relationship, DrugABSTRACT
Development of thermophilic composting for maximizing NH3 gas recovery would enable the production of a nitrogen source which is free from pathogen/heavy metal, for the cultivation of high-value microalgae. The present study examined the effect of NH3 recovery, nitrogen mass balance, and microbial community dynamics on thermophilic composting of shrimp aquaculture sludge. The emission of NH3 gas at 60 and 70⯰C was 14.7% and 15.6%, respectively, which was higher than that at 50⯰C (9.0%). The nitrogen mass balance analysis revealed that higher temperatures enhanced the solubilization of non-dissolved nitrogen and liberation of NH3 gas from the produced NH4+-N. High-throughput microbial community analysis revealed the shift of the dominant bacterial group from Bacillus to Geobacillus with the rise of composting temperature. In conclusion, thermophilic composting of shrimp aquaculture sludge at 60-70⯰C was the most favorable condition for enhancing NH3 gas recovery.
Subject(s)
Aquaculture , Composting , Nitrogen/chemistry , Sewage , Ammonia , Soil , TemperatureABSTRACT
Microalgae are promising candidate resources from marine ecology for health-improving effects. Metabolite profiling of the microalgal diatom, Chaetoceros calcitrans was conducted by using robust metabolomics tools, namely ¹H nuclear magnetic resonance (NMR) spectroscopy coupled with multivariate data analysis (MVDA). The unsupervised data analysis, using principal component analysis (PCA), resolved the five types of extracts made by solvents ranging from polar to non-polar into five different clusters. Collectively, with various extraction solvents, 11 amino acids, cholesterol, 6 fatty acids, 2 sugars, 1 osmolyte, 6 carotenoids and 2 chlorophyll pigments were identified. The fatty acids and both carotenoid pigments as well as chlorophyll, were observed in the extracts made from medium polar (acetone, chloroform) and non-polar (hexane) solvents. It is suggested that the compounds were the characteristic markers that influenced the separation between the clusters. Based on partial least square (PLS) analysis, fucoxanthin, astaxanthin, violaxanthin, zeaxanthin, canthaxanthin, and lutein displayed strong correlation to 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and nitric oxide (NO) inhibitory activity. This metabolomics study showed that solvent extractions are one of the main bottlenecks for the maximum recovery of bioactive microalgal compounds and could be a better source of natural antioxidants due to a high value of metabolites.
Subject(s)
Antioxidants/metabolism , Diatoms/metabolism , Microalgae/metabolism , Nitric Oxide/metabolism , Amino Acids/metabolism , Carotenoids/metabolism , Chlorophyll/metabolism , Cholesterol/metabolism , Fatty Acids/metabolism , Metabolomics/methods , Proton Magnetic Resonance Spectroscopy/methods , Solvents/metabolismABSTRACT
The responses of two species of microalgae, Chlorella sorokiniana and Chlorella zofingiensis, were compared regarding their morphological and biochemical properties under photoautotrophic and mixotrophic conditions. These microalgae were cultured under both conditions, and their crude ethanolic extracts were examined for their pigment and total phenolic contents. In addition, the microalgae's antioxidant activities were determined using a DPPH radical scavenging assay and a ferric reducing antioxidant power (FRAP) assay. Both strains showed increases in cell size due to the accumulation of lipid bodies and other cell contents, especially carotenoids, under the mixotrophic condition. Notably, reductions in phenolic and chlorophyll contents were observed to be associated with lower antioxidant activity. C. zofingiensis compared with C. sorokiniana, demonstrated higher antioxidant activity and carotenoid content. This study showed that different species of microalgae responded differently to varying conditions by producing different types of metabolites, as evidenced by the production of higher levels of phenolic compounds under the photoautotrophic condition and the production of the same levels of carotenoids under both photoautotrophic and mixotrophic conditions.
ABSTRACT
Isoforskolin (ISOF) has been reported to play an important role in many illnesses including respiratory, cardiovascular and ophthalmologic diseases. In our study, we aimed to investigate how ISOF regulates airway remodeling and inflammation in asthma. Based on SO2-stimulated mouse cough model, we assessed the role of ISOF in cough and secretion of phlegm. Afterwards, platelet derived growth factor (PDGF)-induced primary rat airway smooth muscle cell (ASMC) model and ovalbumin (OVA)-induced rat asthma model were used to continue our following research. Our results showed that ISOF could prolong the cough latent period, reduce the cough times in two minutes, and increase the excretion of red phenol, which suggested the antitussive and expectorant effects of ISOF. Besides, ISOF pretreatment reversed the hypotonicity and cytoskeleton remodeling in PDGF-induced ASMCs, and reduced mucus hypersecretion and collagen overdeposition in OVA-induced rat asthma model, which indicated its inhibition on airway remodeling in vitro and in vivo. Moreover, ISOF reduced the invasion of inflammatory cells into bronchoalveolar lavage fluid (BALF) and lungs, which revealed its inhibitory role in airway inflammation. The down-regulation of transforming growth factor ß1 (TGF-ß1) and interleukin-1ß (IL-1ß) upon ISOF treatment might be responsible for its anti-remodeling and anti-inflammation roles. In conclusion, ISOF can reduce cough and sputum, as well as inhibit airway remodeling and inflammation by regulating the expression of TGF-ß1 and IL-1ß. These data indicate the potency of ISOF in treating asthma and also provide insights into the development of new anti-asthma agent.
Subject(s)
Airway Remodeling/drug effects , Asthma/prevention & control , Colforsin/pharmacology , Myocytes, Smooth Muscle/drug effects , Ovalbumin/toxicity , Platelet-Derived Growth Factor/toxicity , Airway Remodeling/physiology , Animals , Anti-Asthmatic Agents/pharmacology , Anti-Asthmatic Agents/therapeutic use , Asthma/chemically induced , Asthma/pathology , Colforsin/analogs & derivatives , Disease Models, Animal , Dose-Response Relationship, Drug , Inflammation/chemically induced , Inflammation/pathology , Inflammation/prevention & control , Male , Mice , Myocytes, Smooth Muscle/pathology , Random Allocation , Rats , Rats, Sprague-DawleyABSTRACT
Partially myristoylated chitosan pyrrolidone carboxylate (PMCP) is a cationic amphiphilic chitosan derivative. Glabridin (Glab) from licorice root extracts is a hydrophobic antimelanogenic agent. Here we assessed the effects of cationic Glab-containing polymeric micelles derived from PMCP (Glab/PMCP-PM) on the ability of Glab to penetrate the skin and inhibit melanogenesis using a human skin model. The amount of Glab absorbed 24 h after the application of Glab/PMCP-PM was approximately four times higher than that of conventional oil-in-water micelles (control) prepared using Tween 60. Further, the release of IL-1α, a mediator of inflammation, was not detected. Treatment with Glab/PMCP-PM significantly increased the inhibition of melanogenesis compared with control. The inhibition of melanogenesis depends upon the enhanced ability of Glab to penetrate the skin, particularly the epidermis. Moreover, the inhibition of melanogenesis and the cationic potential of the Glab/PMCP-PM levels were increased by the cationic phospholipid copolymer. Therefore, Glab/PMCP-PM shows potential as an effective transdermal delivery system for treating skin hyperpigmentation.
Subject(s)
Cations , Chitosan , Drug Carriers , Isoflavones/administration & dosage , Micelles , Phenols/administration & dosage , Administration, Cutaneous , Cations/chemistry , Chitosan/chemistry , Drug Carriers/chemistry , Humans , Hydrophobic and Hydrophilic Interactions , Isoflavones/chemistry , Melanocytes/drug effects , Melanocytes/metabolism , Phenols/chemistry , Polymers/chemistry , Skin/drug effects , Skin/metabolism , Skin AbsorptionABSTRACT
MiRNAs may promote or inhibit tumor recurrence and drug resistance. MiR-139-5p is reportedly downregulated in colorectal cancer patient samples, but it is unknown whether and how miR-139-5p regulates drug resistance. Cancer stem cells (CSCs) are postulated to be important promoters of multiple drug resistance (MDR). In this study, we established a MDR cell model which strongly expressed the CSC-associated biomarkers CD44 and CD133. MiR-139-5p expression was reduced in MDR cell lines, while overexpression of miR-139-5p reversed CD44+/CD133+-associated MDR. We also identified NOTCH1, an important protein for stem cell maintenance and function, as a direct target of miR-139-5p, both in vitro and in a knockout mouse model. Notch1 expression was upregulated in tumor samples and inversely correlated with expression of miR-139-5p. Silencing NOTCH1 exerted an effect similar to overexpression of miR-139-5p by inhibiting the CD44+ and CD133+ population and reversing the drug-resistant phenotype. In conclusion, miR-139-5p downregulated NOTCH1 signaling to reverse CD44+/CD133+-associated MDR in colorectal cancer cells. Given this insight into the miRNA regulation of MDR, miR-139-5p could be a promising therapeutic target for colorectal cancer therapy.
Subject(s)
Colorectal Neoplasms/genetics , Drug Resistance, Multiple/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , RNA Interference , Receptor, Notch1/genetics , AC133 Antigen/metabolism , Animals , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Colorectal Neoplasms/metabolism , Disease Models, Animal , Humans , Hyaluronan Receptors/metabolism , Inhibitory Concentration 50 , Mice , Neoplastic Stem Cells/drug effects , Neoplastic Stem Cells/metabolism , Xenograft Model Antitumor AssaysABSTRACT
microRNA-27a (miR-27a) is frequently dysregulated in human carcinoma, including gastric cancer. The B-cell translocation gene 2 (BTG2) has been implicated in gastric carcinogenesis. However, till now, the link between miR-27a and BTG2 in gastric cancer has not been reported. Here, we found that two isoforms of mature miR-27a, miR-27a-5p and miR-27-3p, were both frequently overexpressed in gastric cancer tissues and cell lines, whereas the expression level of miR-27-3p in gastric cancer was significantly higher than that of miR-27a-5p. And overexpression of miR-27a-3p, but not miR-27a-5p, markedly promoted gastric cancer cell proliferation in vitro as well as tumor growth in vivo. Further experiments revealed that BTG2 was a direct and functional target of miR-27a-3p in gastric cancer and miR-27a-3p inhibition obviously up-regulated the expression of BTG2. In turn, overexpression of BTG2 triggered G1/S cell cycle arrest, induced subsequent apoptosis, and inhibited C-myc activation following Ras/MEK/ERK signaling pathway, which involved in the biological effects of miR-27a-3p/BTG2 axis on gastric carcinogenesis and cancer progression. Overall, these results suggested that the miR-27a-3p/BTG2 axis might represent a promising diagnostic biomarker for gastric cancer patients and could be a potential therapeutic target in the management of gastric cancer.
Subject(s)
Gene Expression Regulation, Neoplastic/genetics , Immediate-Early Proteins/biosynthesis , MicroRNAs/genetics , Stomach Neoplasms/genetics , Tumor Suppressor Proteins/biosynthesis , Animals , Heterografts , Humans , Immediate-Early Proteins/genetics , Mice , Mice, Nude , Oncogenes , Tumor Suppressor Proteins/geneticsABSTRACT
The recalcitrant landfill leachate was anaerobically digested at various mixing ratios with labile synthetic wastewater to evaluate the degradation properties of recalcitrant wastewater. The proportion of leachate to the digestion system was increased in three equal steps, starting from 0% to 100%, and later decreased back to 0% with the same steps. The chemical oxygen demand (COD) for organic carbon and other components were calculated by analyzing the COD and dissolved organic carbon (DOC), and the removal efficiencies of COD carbon and COD others were evaluated separately. The degradation properties of COD carbon and COD others shifted owing to changing of substrate degradability, and the removal efficiencies of COD carbon and COD others were improved after supplying 100% recalcitrant wastewater. The UV absorptive property and total organic carbon (TOC) of each molecular size using high performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with UVA and TOC detectors were also investigated, and the degradability of different molecular sizes was determined. Although the SEC system detected extracellular polymeric substances (EPS), which are produced by microbes in stressful environments, during early stages of the experiment, EPS were not detected after feeding 100% recalcitrant wastewater. These results suggest that the microbes had acclimatized to the recalcitrant wastewater degradation. The high removal rates of both COD carbon and COD others were sustained when the proportion of labile wastewater in the substrate was 33%, indicating that the effective removal of recalcitrant COD might be controlled by changing the substrate's degradability.
Subject(s)
Waste Disposal, Fluid/methods , Water Pollutants, Chemical , Anaerobiosis , Biodegradation, Environmental , Biological Oxygen Demand Analysis , Bioreactors , Carbon/analysis , Carbon/metabolism , Chromatography, Gel/methods , Chromatography, High Pressure Liquid/methods , Wastewater/chemistry , Water Pollutants, Chemical/chemistry , Water Pollutants, Chemical/metabolismABSTRACT
To determine the optimum light intensity per cell required for rapid growth regardless of cell density, continuous cultures of the microalga Chlorella zofingiensis were grown with a sufficient supply of nutrients and CO2 and were subjected to different light intensities in the range of 75-1000 µE m(-2) s(-1). The cell density of culture increased over time for all light conditions except for the early stage of the high light condition of 1000 µE m(-2) s(-1). The light intensity per cell required for the high specific growth rate of 0.5 day(-1) was determined to be 28-45 µE g-ds(-1) s(-1). The specific growth rate was significantly correlated to light intensity (y=0.721×x/(66.98+x), r(2)=0.85, p<0.05). A high specific growth rate was maintained over a range of light intensities (250-1000 µE m(-2) s(-1)). This range of light intensities suggested that effective production of C. zofingiensis can be maintained outdoors under strong light by using the optimum specific light intensity.