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1.
Fitoterapia ; 172: 105758, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38042507

ABSTRACT

The Chinese herb Qianghuo is an antiphlogistic herb with many effects and complex components. In this study, the chemical composition of Qianghuo and its components in rat plasma after oral administration were investigated using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). The extracts, blank plasma, and plasma containing the drug were analyzed by mass spectrometry, and data collected in both positive and negative ion modes were analyzed using Masslynx software, and the structures were confirmed by combining the compound fragment ions and mass spectrometry cleavage pathways. A total of 62 in vitro chemical components were identified, including 27 coumarins, 18 organic acids, 5 amino acids, 5 glycosides, 2 flavonoids, 4 nucleotides, and 1 ester, which were summarized from the obtained compounds in terms of their possible cleavage patterns. Among the identified 31 compounds in rat plasma, 21 were prototypes, mostly coumarins, organic acids, and flavonoids, and 10 were metabolites, which were mainly generated via hydroxylation and methylation pathways. Based on these, this study provides a theoretical foundation for quality control and basic research on Qianghuo medicinal substances.


Subject(s)
Drugs, Chinese Herbal , Tandem Mass Spectrometry , Rats , Animals , Tandem Mass Spectrometry/methods , Drugs, Chinese Herbal/chemistry , Chromatography, High Pressure Liquid/methods , Molecular Structure , Flavonoids/analysis , Acids , Coumarins/analysis
2.
Fish Shellfish Immunol ; 135: 108653, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36868540

ABSTRACT

Echinacea purpurea (Linn.) Moench (EP) is a globally popular herbal medicine, which showed effects on growth promotion, antioxidant and immunomodulatory activities in fish culture world widely. However, there are few studies about the effects on miRNAs by EP in fish. The hybrid snakehead fish (Channa maculate♀ × Channa argus ♂) was new important economic specie of freshwater aquaculture in China with high market value and demand while there were only a few reports about its miRNAs. To overview immune-related miRNAs of the hybrid snakehead fish and to further understand the immune regulating mechanism of EP, we herein constructed and analyzed three small RNA libraries of immune tissues including liver, spleen and head kidney of the fish with or without EP treatment via Illumina high-throughput sequencing technology. Results showed that EP can affect the immune activities of fish by the miRNA-regulated ways. Totally, 67 (47 up and 20 down) miRNAs in liver, 138 (55 up and 83 down) miRNAs in spleen, and 251 (15 up and 236 down) miRNAs in spleen were detected, as well as 30, 60, 139 kinds of immune-related miRNAs belonging to 22, 35 and 66 families of the three tissues respectively. The expressions of 8 immune-related miRNA family members were found in all the three tissues, including miR-10, miR-133, miR-22 and etc. Some miRNAs have been identified involved in the innate and adaptive immune responses, such as the miR-125, miR-138, and miR-181 family. Ten miRNA families with antioxidant target genes were also discovered, including miR-125, miR-1306, and miR-138, etc. Results from Gene Ontology (GO) and KEGG pathway analysis further confirmed there are a majority immune response targets of the miRNAs involved in the EP treatment process. Our study deepened understanding roles of miRNAs in fish immune system and provides new ideas for the study of immune mechanism of EP.


Subject(s)
Echinacea , MicroRNAs , Animals , MicroRNAs/genetics , MicroRNAs/metabolism , Antioxidants , Fishes
3.
Cell Rep ; 42(2): 112102, 2023 02 28.
Article in English | MEDLINE | ID: mdl-36774548

ABSTRACT

Nutritional symbionts influence host reproduction, but the underlying molecular mechanisms are largely unclear. We previously found that the bacteriocyte symbiont Hamiltonella impacts the sex ratio of the whitefly Bemisia tabaci. Hamiltonella synthesizes folate by cooperation with the whitefly. Folate deficiency by Hamiltonella elimination or whitefly gene silencing distorted whitefly sex ratio, and folate supplementation restored the sex ratio. Hamiltonella deficiency or gene silencing altered histone H3 lysine 9 trimethylation (H3K9me3) level, which was restored by folate supplementation. Genome-wide chromatin immunoprecipitation-seq analysis of H3K9me3 indicated mitochondrial dysfunction in symbiont-deficient whiteflies. Hamiltonella deficiency compromised mitochondrial quality of whitefly ovaries. Repressing ovary mitochondrial function led to distorted whitefly sex ratio. These findings indicate that the symbiont-derived folate regulates host histone methylation modifications, which thereby impacts ovary mitochondrial function, and finally determines host sex ratio. Our study suggests that a nutritional symbiont can regulate animal reproduction in a way that differs from reproductive manipulators.


Subject(s)
Hemiptera , Animals , Female , Hemiptera/genetics , Sex Ratio , Symbiosis/genetics , Enterobacteriaceae/genetics , Folic Acid
4.
Appl Environ Microbiol ; 88(3): e0208921, 2022 02 08.
Article in English | MEDLINE | ID: mdl-34818107

ABSTRACT

Nutritional symbionts are restricted to specialized host cells called bacteriocytes in various insect orders. These symbionts can provide essential nutrients to the host. However, the cellular mechanisms underlying the regulation of these insect-symbiont metabolic associations remain largely unclear. The whitefly Bemisia tabaci MEAM1 hosts "Candidatus Portiera aleyrodidarum" (here, "Ca. Portiera") and "Candidatus Hamiltonella defensa" (here, "Ca. Hamiltonella") bacteria in the same bacteriocyte. In this study, the induction of autophagy by chemical treatment and gene silencing decreased symbiont titers and essential amino acid (EAA) and B vitamin contents. In contrast, the repression of autophagy in bacteriocytes via Atg8 silencing increased symbiont titers, and amino acid and B vitamin contents. Furthermore, dietary supplementation with non-EAAs or B vitamins alleviated autophagy in whitefly bacteriocytes, elevated TOR (target of rapamycin) expression, and increased symbiont titers. TOR silencing restored symbiont titers in whiteflies after dietary supplementation with B vitamins. These data suggest that "Ca. Portiera" and "Ca. Hamiltonella" evade autophagy of the whitefly bacteriocytes by activating the TOR pathway via providing essential nutrients. Taken together, we demonstrate that autophagy plays a critical role in regulating the metabolic interactions between the whitefly and two intracellular symbionts. Therefore, this study reveals that autophagy is an important cellular basis for bacteriocyte evolution and symbiosis persistence in whiteflies. The whitefly symbiosis unravels the interactions between cellular and metabolic functions of bacteriocytes. IMPORTANCE Nutritional symbionts, which are restricted to specialized host cells called bacteriocytes, can provide essential nutrients for many hosts. However, the cellular mechanisms of regulation of animal-symbiont metabolic associations have been largely unexplored. Here, using the whitefly-"Ca. Portiera"/"Ca. Hamiltonella" endosymbiosis, we demonstrate autophagy regulates the symbiont titers and thereby alters the essential amino acid and B vitamin contents. For persistence in the whitefly bacteriocytes, "Ca. Portiera" and "Ca. Hamiltonella" alleviate autophagy by activating the TOR (target of rapamycin) pathway through providing essential nutrients. Therefore, we demonstrate that autophagy plays a critical role in regulating the metabolic interactions between the whitefly and two intracellular symbionts. This study also provides insight into the cellular basis of bacteriocyte evolution and symbiosis persistence in the whitefly. The mechanisms underlying the role of autophagy in whitefly symbiosis could be widespread in many insect nutritional symbioses. These findings provide a new avenue for whitefly control via regulating autophagy in the future.


Subject(s)
Halomonadaceae , Hemiptera , Vitamin B Complex , Animals , Autophagy , Halomonadaceae/genetics , Hemiptera/microbiology , Symbiosis/genetics , Vitamin B Complex/metabolism
5.
Medicine (Baltimore) ; 100(44): e27503, 2021 Nov 05.
Article in English | MEDLINE | ID: mdl-34871212

ABSTRACT

BACKGROUND: Acne is a common inflammatory disease of sebaceous glands, which brings extensive emotional and psychological distress to patients. Although acupuncture has certain advantages in the treatment of acne, the curative effect is not exact. The purpose of this trial is to evaluate the feasibility, preliminary efficacy, and safety of the "Spleen and Stomach Guiyuan Acupuncture Method" (SSGA) in the treatment of gastrointestinal damp-heat acne. METHODS: The proposed protocol is planned as a randomized, assessor-blind, conventional-treatment-controlled trial to evaluate the efficacy of SSGA on gastrointestinal damp-heat acne. Seventy six gastrointestinal damp-heat acne patients will be randomly divided into 2 groups and receive SSGA or conventional acupuncture treatment. The entire study period is 12 weeks, including an 8-week treatment period and a 4-week follow-up period. All patients will receive 16 sessions of acupuncture treatment over 8 weeks. The primary outcome is the investigation global assessment (IGA) at week 8, which is an overall assessment of the degree of the inflammatory and non-inflammatory lesion. The secondary outcomes include IGA, the total facial lesion count (Total Lesion Count), the acne-specific quality of life, etc at weeks 8 and 12. The Expectation and Credibility of treatment rating scale will be used to measure the patients' attitudes to acupuncture after the first treatment. Adverse events will also be recorded. DISCUSSION: This study is helpful to evaluate the feasibility, preliminary efficacy, and safety of SSGA in the treatment of gastrointestinal damp-heat acne. The results will be used in sample size calculations for subsequent large-scale studies.Trial registration: Chinese Clinical Trial Registry ChiCTR2100047363. Registered on June 13, 2021.


Subject(s)
Acne Vulgaris/therapy , Acupuncture Therapy , Acne Vulgaris/psychology , Hot Temperature , Humans , Immunoglobulin A , Quality of Life , Randomized Controlled Trials as Topic , Treatment Outcome
6.
ISME J ; 15(6): 1655-1667, 2021 06.
Article in English | MEDLINE | ID: mdl-33432136

ABSTRACT

Intracellular symbionts in insects often have reduced genomes. Host acquisition of genes from bacteria is an important adaptation that supports symbionts. However, the function of horizontally transferred genes in insect symbiosis remains largely unclear. The primary symbiont Portiera housed in bacteriocytes lacks pantothenate synthesis genes: panB and panC, which is presumably complemented by a fused gene panB-panC (hereafter panBC) horizontally transferred from bacteria in Bemisia tabaci MEAM1. We found panBC in many laboratory cultures, and species of B. tabaci shares a common evolutionary origin. We demonstrated that complementation with whitefly panBC rescued E. coli pantothenate gene knockout mutants. Portiera elimination decreased the pantothenate level and PanBC abundance in bacteriocytes, and reduced whitefly survival and fecundity. Silencing PanBC decreased the Portiera titer, reduced the pantothenate level, and decreased whitefly survival and fecundity. Supplementation with pantothenate restored the symbiont titer, PanBC level, and fitness of RNAi whiteflies. These data suggest that pantothenate synthesis requires cooperation and coordination of whitefly PanBC expression and Portiera. This host-symbiont co-regulation was mediated by the pantothenate level. Our findings demonstrated that pantothenate production, by the cooperation of a horizontally acquired, fused bacteria gene and Portiera, facilitates the coordination of whitefly and symbiont fitness. Thus, this study extends our understanding on the basis of complex host-symbiont interactions.


Subject(s)
Hemiptera , Vitamin B Complex , Animals , Bacteria/genetics , Escherichia coli , Symbiosis
7.
ISME J ; 14(12): 2923-2935, 2020 12.
Article in English | MEDLINE | ID: mdl-32690936

ABSTRACT

Symbionts can regulate animal reproduction in multiple ways, but the underlying physiological and biochemical mechanisms remain largely unknown. The presence of multiple lineages of maternally inherited, intracellular symbionts (the primary and secondary symbionts) in terrestrial arthropods is widespread in nature. However, the biological, metabolic, and evolutionary role of co-resident secondary symbionts for hosts is poorly understood. The bacterial symbionts Hamiltonella and Arsenophonus have very high prevalence in two globally important pests, the whiteflies Bemisia tabaci and Trialeurodes vaporariorum, respectively. Both symbionts coexist with the primary symbiont Portiera in the same host cell (bacteriocyte) and are maternally transmitted. We found that elimination of both Hamiltonella and Arsenophonous by antibiotic treatment reduced the percentage of female offspring in whiteflies. Microsatellite genotyping and cytogenetic analysis revealed that symbiont deficiency inhibited fertilization in whiteflies, leading to more haploid males with one maternal allele, which is consistent with distorted sex ratio in whiteflies. Quantification of essential amino acids and B vitamins in whiteflies indicated that symbiont deficiency reduced B vitamin levels, and dietary B vitamin supplementation rescued fitness of whiteflies. This study, for the first time, conclusively demonstrates that these two intracellular symbionts affect sex ratios in their whitefly hosts by regulating fertilization and supplying B vitamins. Our results reveal that both symbionts have the convergent function of regulating reproduction in phylogenetically-distant whitefly species. The 100% frequency, the inability of whiteflies to develop normally without their symbiont, and rescue with B vitamins suggests that both symbionts may be better considered co-primary symbionts.


Subject(s)
Hemiptera , Vitamin B Complex , Animals , Female , Fertilization , Male , Sex Ratio , Symbiosis
8.
ISME J ; 14(10): 2542-2553, 2020 10.
Article in English | MEDLINE | ID: mdl-32572143

ABSTRACT

Insect symbionts are widespread in nature and lateral gene transfer is prevalent in insect symbiosis. However, the function of horizontally transferred genes (HTGs) in insect symbiosis remains speculative, including the mechanism that enables insects to feed on plant phloem deficient in B vitamins. Previously, we found there is redundancy in biotin synthesis pathways from both whitefly Bemisia tabaci and symbiotic Hamiltonella due to the presence of whitefly HTGs. Here, we demonstrate that elimination of Hamiltonella decreased biotin levels but elevated the expression of horizontally transferred biotin genes in whiteflies. HTGs proteins exhibit specific expression patterns in specialized insect cells called bacteriocytes housing symbionts. Complementation with whitefly HTGs rescued E. coli biotin gene knockout mutants. Furthermore, silencing whitefly HTGs in Hamiltonella-infected whiteflies reduced biotin levels and hindered adult survival and fecundity, which was partially rescued by biotin supplementation. Each of horizontally transferred biotin genes are conserved in various laboratory cultures and species of whiteflies with geographically diverse distributions, which shares an evolutionary origin. We provide the first experimental evidence that biotin synthesized through acquired HTGs is important in whiteflies and may be as well in other animals. Our findings suggest that B vitamin provisioning in animal-microbe symbiosis frequently evolved from bacterial symbionts to animal hosts through horizontal gene transfer events. This study will also shed light on how the animal genomes evolve through functional transfer of genes with bacterial origin in the wider contexts of microbial ecology.


Subject(s)
Biotin , Hemiptera , Animals , Bacteria/genetics , Escherichia coli , Symbiosis
9.
Phytochem Anal ; 31(6): 739-746, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32304155

ABSTRACT

INTRODUCTION: Vinca alkaloids are important sources for producing anticancer drugs from Catharanthus roseus. The phosphorus of soil is one of crucial factors for planting C. roseus. OBJECTIVES: We aim to develop an in vivo sampling technique coupled with direct mass spectrometry with wooden tip for investigating distributions and changes of alkaloids in flowers, leaves, stems, veins and roots of living C. roseus under low-phosphorus stress. MATERIALS AND METHODS: Living C. roseus were prepared under low-phosphorus stress (n = 10) and control conditions (n = 10). Wooden-tip electrospray ionisation mass spectrometry and conventional liquid chromatography-mass spectrometry were applied to analyse living C. roseus and extracts of C. roseus, respectively. RESULTS: Distributions and changes of serpentine, vindoline, catharanthine, and anhydrovinblastine in living C. roseus under low-phosphorus stress and control conditions were successfully obtained. CONCLUSION: Compared to control soil conditions, low-phosphorus soil was found to induce C. roseus to generate more serpentine but less catharanthine and vindoline in leaves, veins, stems and roots, and to generate more anhydrovinblastine in flowers, leaves, stems and roots. Overall, our results showed a simple, rapid, and effective method for in vivo sampling and direct analysis of living plants.


Subject(s)
Catharanthus , Chromatography, High Pressure Liquid , Phosphorus , Plant Leaves , Spectrometry, Mass, Electrospray Ionization
10.
Rapid Commun Mass Spectrom ; 33(24): 1877-1883, 2019 Dec 30.
Article in English | MEDLINE | ID: mdl-31429145

ABSTRACT

RATIONALE: Herbal dietary supplements (HDSs) adulterated with undeclared synthetic drugs can lead to serious health problems METHODS: A fast-switching positive/negative high-voltage (+/- HV) was developed to apply on electrospray ionization mass spectrometry (ESI-MS) with porous tips for rapid screening of five antirheumatic drugs in antirheumatic HDSs. The fast-switching (switch-time: 100 ms) negative and positive ions were alternately generated to perform full-MS and tandem-MS analysis, providing an effective method for rapid detection of analytes in whichever mode of detection was most suitable (negative or positive ion mode). The use of different tips and solvents was also optimized in this work. RESULTS: The limits of detection of the five antirheumatic drugs were found to be less than 0.1 ng/g (S/N > 3). The reproducibility of the five drugs was measured to be 10.0-23.3% (n = 5). A single sample analysis could be completed within 1 min. Rapid screening of a total of 28 real HDS samples collected from the market was examined by the fast-switching HV substrate-tip ESI-MS method, and the screening result was further validated by conventional liquid chromatography/mass spectrometry. CONCLUSIONS: Overall, our results demonstrated that fast-switching HV substrate-tip ESI-MS is a rapid, reliable, and effective method for simultaneous screening of various analytes in complex samples.


Subject(s)
Antirheumatic Agents/analysis , Drug Contamination , Drugs, Chinese Herbal/analysis , Spectrometry, Mass, Electrospray Ionization/methods , Dietary Supplements/analysis , Drug Contamination/statistics & numerical data , Sensitivity and Specificity
11.
J Dermatolog Treat ; 30(6): 605-611, 2019 Sep.
Article in English | MEDLINE | ID: mdl-30388912

ABSTRACT

Objective: Oral lichen planus (OLP) is a relatively common immunological mucocutaneous disease that causes pain and poor quality of life. Curcumin has been reported to be a safe and effective treatment for OLP. The objective of this review is to evaluate the existing evidence for the safety of curcumin in treating OLP as well as its efficacy compared with that of corticosteroids. Methods: We reviewed the published literature by searching PubMed, EMBASE, EBSCO, and Cochrane Library, and then retrieved and analyzed several variables from patient records. Results: Nine studies met the inclusion criteria, including six randomized, double-blind clinical trials; two pilot clinical trials; and one case report. A total of 259 OLP patients were included in the systematic review. Seven studies showed statistically significant differences in pain severity and clinical appearance of oral lesions after treatment with curcumin for a period of time, compared to baseline (p < .05). Three controlled clinical trials compared the efficacy of curcumin to that of corticosteroids; all of these trials showed no statistically significant differences in pain severity and clinical appearance of oral lesions. Conclusions: Curcumin is a safe treatment and can be used as an adjunct in combination with corticosteroids to reduce pain, burning sensations, and the clinical appearance of oral lesions in OLP patients.


Subject(s)
Curcumin/therapeutic use , Lichen Planus, Oral/drug therapy , Adrenal Cortex Hormones/therapeutic use , Clinical Trials as Topic , Curcumin/adverse effects , Databases, Factual , Gastrointestinal Diseases/etiology , Humans , Quality of Life , Treatment Outcome
12.
Qual Life Res ; 26(4): 1007-1014, 2017 04.
Article in English | MEDLINE | ID: mdl-27696109

ABSTRACT

PURPOSE: This study aimed to test the effects of horticulture therapy on activities of daily living, happiness, meaning of life, and interpersonal intimacy of nursing home older adults in southern Taiwan. METHODS: A quasi-experimental study was applied. Eighty-five older adults aged 65 or older who lived in nursing homes in southern Taiwan were recruited conveniently. All participants completed the study: experimental group (n = 41) and control group (n = 44). The experimental group received horticulture therapy for 1 h once a week for 8 weeks, while the control group continued their routine daily activities. The following questionnaires were administered before and after the intervention period: (1) Barthel Index (BI), (2) Chinese Happiness Inventory short version (CHI), (3) Meaning of Life Scale (MLS), and (4) Interpersonal Intimacy Scale (IIS). RESULTS: The BI, CHI, MLS, and IIS scores significantly improved in the experimental group (p < .05). After 8 weeks of horticulture therapy, the BI, CHI, and IIS scores of experimental group participants were significantly better than the scores of control group participants (p < .05); however, the MLS scores of two groups showed no significant differences (p = .738). CONCLUSIONS: Horticulture therapy improved activities of daily living, happiness, and interpersonal intimacy of older adults in nursing homes. We recommend that nursing homes recruit and train personnel to lead horticultural therapy and to incorporate the therapy as routine daily activities in the facilities.


Subject(s)
Activities of Daily Living , Frail Elderly/psychology , Horticultural Therapy , Nursing Homes , Quality of Life , Aged , Aged, 80 and over , Female , Health Services for the Aged , Humans , Male , Surveys and Questionnaires , Taiwan
13.
J Ethnopharmacol ; 189: 132-8, 2016 Aug 02.
Article in English | MEDLINE | ID: mdl-27196296

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: The Chinese herb compound prescription Viola yedoensis Makino Anti-itching Compound (VYAC), which consists of Viola yedoensis Makino, herb, Sophora flavescens Aiton, root, and Dictamnus dasycarpus Turcz, root and rhizome, has been traditionally used to treat various skin allergic inflammatory diseases in clinic. AIM OF THE STUDY: The aim of this study is to investigate the effects of VYAC on degranulation and to determine its anti-inflammatory mechanism in RBL-2H3 mast cells. MATERIALS AND METHODS: VYAC was extracted with water-coction extraction (Shufen et al., 2012). The aqueous extracts were concentrated in vacuum under reduced pressure and lyophilized using a freeze dryer, and lyophilized powder was obtained. MTT was used to evaluate the cytotoxic of VYAC on RBL-2H3 cells. Degranulation was carried out with RBL-2H3 cell model, which was stimulated with A23187 plus PMA. ß-Hexosaminidase and histamine were measured to evaluate degranulation. The mRNA levels of inflammation cytokines (IL-1ß, TNF-α, IL-6, and iNOS) were investigated by RT-PCR to explain the anti-inflammatory mechanism of VYAC. RESULTS: VYAC did not show cytotoxic effect on RBL-2H3 cells in the range of 25-400µg/mL. A higher dose of VYAC (800µg/mL) showed significant cytotoxicity (P<0.05). VYAC could significantly inhibit ß-hexosaminidase and histamine release when treated with 100, 200, and 400µg/mL (P<0.05), but could not significantly inhibit ß-Hexosaminidase and histamine release when treated with 25 and 50µg/mL (p>0.05). The mRNA levels of inflammatory cytokines (TNF-α, IL-1ß, IL-6, and iNOS) could significantly decrease when treated with 200 and 400µg/mL (P<0.05) of VYAC, which were associated with the development of inflammation. CONCLUSIONS: Results showed that VYAC inhibited ß-hexosaminidase and histamine release, which was inhibit A23187 plus PMA stimulated RBL-2H3 cell degranulation and downregulated inflammatory cytokines (IL-1ß, TNF-α, IL-6, and iNOS) expression to block inflammatory development.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Antipruritics/pharmacology , Cell Degranulation/drug effects , Cytokines/metabolism , Drugs, Chinese Herbal/pharmacology , Histamine/metabolism , Inflammation Mediators/metabolism , Mast Cells/drug effects , Viola/chemistry , Animals , Cell Line, Tumor , Cell Survival/drug effects , Chromatography, High Pressure Liquid , Cytokines/genetics , Cytokines/immunology , Dose-Response Relationship, Drug , Down-Regulation , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Histamine/immunology , Inflammation Mediators/immunology , Mast Cells/immunology , Mast Cells/metabolism , Mast Cells/pathology , Nitric Oxide Synthase Type II/genetics , Nitric Oxide Synthase Type II/metabolism , Phytotherapy , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , beta-N-Acetylhexosaminidases/metabolism
14.
Biochim Biophys Acta ; 1839(7): 579-91, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24852358

ABSTRACT

Aberrant expression levels of transcriptional regulators result in alterations in transcriptional control. STAF65γ is a structural subunit of the GCN5 transcriptional co-activator complex. Reports showed that STAF65γ is highly expressed in several human cancer cells, but the consequences of this aberrant expression pattern remain elusive. Here, we show that the STAF65γ protein is highly expressed in lung adenocarcinoma patients and high levels of STAF65γ correlate with poor prognosis. High levels of STAF65γ cause repression of the c-Myc oncogene through physical association with transcription factor YY1 and co-repressors HDACs. Physical interactions between STAF65γ and class IIa HDACs facilitate nuclear enrichment and regulate the assembly of HDAC complexes. Moreover, SUMOylation of STAF65γ is necessary for maintaining the co-repressor complex containing YY1 and class IIa HDACs at the promoter. Our findings reveal a distinct role of STAF65γ in nuclear import, transcriptional repression, and cell cycle regulation at high levels of expression, which is associated with poor clinical outcomes of lung adenocarcinoma.


Subject(s)
Adenocarcinoma/genetics , Histone Deacetylases/genetics , Lung Neoplasms/genetics , Promoter Regions, Genetic , Trans-Activators/genetics , Transcription, Genetic , Active Transport, Cell Nucleus/genetics , Adenocarcinoma/pathology , Adenocarcinoma of Lung , Adult , Aged , Cell Cycle/genetics , Cell Line, Tumor , Female , Gene Expression Regulation, Neoplastic , Humans , Lung Neoplasms/pathology , Male , Middle Aged , Prognosis , Repressor Proteins/genetics , Sumoylation , YY1 Transcription Factor/genetics
15.
Biochem Biophys Res Commun ; 431(1): 58-64, 2013 Feb 01.
Article in English | MEDLINE | ID: mdl-23291187

ABSTRACT

Poly(ADP-ribose) polymerase-2 (PARP-2) catalyzes poly(ADP-ribosyl)ation (PARylation) and regulates numerous nuclear processes, including transcription. Depletion of PARP-2 alters the activity of transcription factors and global gene expression. However, the molecular action of how PARP-2 controls the transcription of target promoters remains unclear. Here we report that PARP-2 possesses transcriptional repression activity independently of its enzymatic activity. PARP-2 interacts and recruits histone deacetylases HDAC5 and HDAC7, and histone methyltransferase G9a to the promoters of cell cycle-related genes, generating repressive chromatin signatures. Our findings propose a novel mechanism of PARP-2 in transcriptional regulation involving specific protein-protein interactions and highlight the importance of PARP-2 in the regulation of cell cycle progression.


Subject(s)
Cell Cycle/genetics , Gene Expression Regulation , Histones/metabolism , Poly Adenosine Diphosphate Ribose/metabolism , Poly(ADP-ribose) Polymerases/metabolism , G1 Phase/genetics , HEK293 Cells , Histocompatibility Antigens/metabolism , Histone Deacetylases/metabolism , Histone-Lysine N-Methyltransferase/metabolism , Humans , Methylation , Poly(ADP-ribose) Polymerases/genetics , Promoter Regions, Genetic , Proto-Oncogene Proteins c-myc/genetics , YY1 Transcription Factor/metabolism
16.
Genes Dev ; 17(8): 1019-29, 2003 Apr 15.
Article in English | MEDLINE | ID: mdl-12704081

ABSTRACT

Methylation of specific residues within the N-terminal histone tails plays a critical role in regulating eukaryotic gene expression. Although great advances have been made toward identifying histone methyltransferases (HMTs) and elucidating the consequences of histone methylation, little is known about the recruitment of HMTs to regulatory regions of chromatin. Here we report that the sequence-specific DNA-binding transcription factor Yin Yang 1 (YY1) binds to and recruits the histone H4 (Arg 3)-specific methyltransferase, PRMT1, to a YY1-activated promoter. Our data confirm that histone methylation does not occur randomly but rather is a targeted event and provides one mechanism by which HMTs can be recruited to chromatin to activate gene expression.


Subject(s)
Arginine/metabolism , DNA-Binding Proteins/metabolism , Histones/metabolism , Phosphoproteins , Promoter Regions, Genetic/genetics , Protein-Arginine N-Methyltransferases/metabolism , Transcription Factors/metabolism , Amino Acid Sequence , Animals , Binding Sites , Blotting, Western , Chromatin/genetics , DNA Primers/chemistry , DNA-Binding Proteins/genetics , Electrophoretic Mobility Shift Assay , Erythroid-Specific DNA-Binding Factors , Gene Expression Regulation , Glutathione Transferase/chemistry , Glutathione Transferase/metabolism , Luciferases/metabolism , Methylation , Molecular Sequence Data , Nuclear Factor 90 Proteins , Protein-Arginine N-Methyltransferases/genetics , RNA-Binding Proteins/metabolism , Regulatory Sequences, Nucleic Acid , Repressor Proteins , Sequence Deletion , Sequence Homology, Amino Acid , Transcription Factors/genetics , Transcription, Genetic , Transcriptional Activation , Transfection , YY1 Transcription Factor , Zinc Fingers
17.
Mol Cancer Ther ; 1(12): 1079-87, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12481431

ABSTRACT

Selenium compounds are potential chemopreventive agents for prostate cancer. There are several proposed mechanisms for their anticancer effect, including enhanced apoptosis of transformed cells. Because the transcription factor nuclear factor-kappa B (NF-kappa B) is often constitutively activated in tumors and is a key antiapoptotic factor in mammalian cells, we tested whether selenium inhibited NF-kappa B activity in prostate cancer cells. In our work, we used sodium selenite and a novel synthetic compound, methylseleninic acid (MSeA), that served as a precursor of the putative active monomethyl metabolite methylselenol. We found that both selenium forms inhibited cell growth and induced apoptosis in DU145 and JCA1 prostate carcinoma cells. Sodium selenite and MeSeA, at the concentrations that induced apoptosis, inhibited NF-kappa B DNA binding induced by tumor necrosis factor-alpha and lipopolysaccharide in DU145 and JCA1 prostate cells. Both compounds also inhibited kappa B. Luciferase reporter activity in prostate cells. A key to NF-kappa B regulation is the inhibitory kappa B (I kappa B) proteins that in response to diverse stimuli are rapidly phosphorylated by I kappa B kinase complex, ubiquitinated, and undergo degradation, releasing NF-kappa B factor. We showed that sodium selenite and MSeA inhibited I kappa B kinase activation and I kappa B-alpha phosphorylation and degradation induced by TNF-alpha and lipopolysaccharide in prostate cells. NF-kappa B blockage by I kappa B-alpha d.n. mutant resulted in the sensitization of prostate carcinoma cells to apoptosis induced by selenium compounds. These results suggest that selenium may target the NF-kappa B activation pathway to exert, at least in part, its cancer chemopreventive effect in prostate.


Subject(s)
Antineoplastic Agents/pharmacology , Enzyme Inhibitors/pharmacology , Protein Serine-Threonine Kinases/antagonists & inhibitors , Selenium/pharmacology , Active Transport, Cell Nucleus/drug effects , Adenoviridae/genetics , Anticarcinogenic Agents/pharmacology , Apoptosis , Blotting, Western , Cell Nucleus/metabolism , Cytosol/metabolism , Dose-Response Relationship, Drug , Enzyme Activation , Humans , I-kappa B Kinase , Luciferases/metabolism , Male , NF-kappa B/metabolism , Organoselenium Compounds/pharmacology , Poly(ADP-ribose) Polymerases/metabolism , Prostatic Neoplasms/metabolism , Protein Binding , Time Factors , Transcription, Genetic , Transfection , Tumor Cells, Cultured
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