1 4993 177 PENTOXIFYLLINE-INDUCED PROTEIN EXPRESSION CHANGE IN RAW 264.7 CELLS AS DETERMINED BY IMMUNOPRECIPITATION-BASED HIGH PERFORMANCE LIQUID CHROMATOGRAPHY. ALTHOUGH PENTOXIFYLLINE (PTX) WAS IDENTIFIED AS A COMPETITIVE NON-SELECTIVE PHOSPHODIESTERASE INHIBITOR, ITS PHARMACOLOGICAL EFFECT HAS NOT BEEN CLEARLY ELUCIDATED. THE PRESENT STUDY EXPLORED THE EFFECT OF LOW DOSE 10 MUG/ML PTX (THERAPEUTIC DOSE) COMPARED TO HIGH DOSE 300 MUG/ML PTX (EXPERIMENTAL DOSE) IN RAW 264.7 CELLS THROUGH IMMUNOPRECIPITATION-BASED HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (IP-HPLC), IMMUNOHISTOCHEMISTRY, AND WESTERN BLOT. 10 MUG/ML PTX INCREASED THE EXPRESSION OF PROLIFERATION (KI-67, PCNA, CYCLIN D2, CDC25A), EPIGENETIC MODIFICATION (KDM4D, PCAF, HMGB1), PROTEIN TRANSLATION (DOHH, DHPS, EIF5A1), RAS SIGNALING (KRAS, PAKT1/2/3, PI3K), NFKB SIGNALING (NFKB, GADD45, P38), PROTECTION (HSP70, SOD1, GSTO1/2), SURVIVAL (PAKT1/2/3, SP1, SIRTUIN 6), NEUROMUSCULAR DIFFERENTIATION (NSEGAMMA, MYOSIN-1A, DESMIN), OSTEOBLASTIC DIFFERENTIATION (BMP2, RUNX2, OSTERIX), ACUTE INFLAMMATION (TNFALPHA, IL-1, CXCR4), INNATE IMMUNITY (BETA-DEFENSIN 1, LACTOFERRIN, TLR-3, -4), CELL-MEDIATED IMMUNITY (CD4, CD8, CD80), WHILE DECREASED THE EXPRESSION OF ER STRESS (EIF2ALPHA, EIF2AK3, ATF6ALPHA), FIBROSIS (FGF2, CTGF, COLLAGEN 3A1), AND CHRONIC INFLAMMATION (CD68, MMP-2, -3, COX2) VERSUS THE UNTREATED CONTROLS. THE ACTIVATION OF PROLIFERATION BY 10 MUG/ML PTX WAS ALSO SUPPORTED BY THE INCREASE OF CMYC-MAX HETERODIMER AND BETA-CATENIN-TCF1 COMPLEX IN DOUBLE IP-HPLC. 10 MUG/ML PTX ENHANCED FAS-MEDIATED APOPTOSIS BUT DIMINISHED P53-MEDIATED APOPTOSIS, AND DOWNREGULATED MANY ANGIOGENESIS PROTEINS (ANGIOGENIN, VEGF-A, AND FLT4), BUT UPREGULATED HIF1ALPHA, VEGFR2, AND CMG2 REACTIVELY. WHEREAS, 300 MUG/ML PTX CONSISTENTLY DECREASED PROLIFERATION, EPIGENETIC MODIFICATION, RAS AND NFKB SIGNALING, NEUROMUSCULAR AND OSTEOBLASTIC DIFFERENTIATION, BUT INCREASED APOPTOSIS, ER STRESS, AND FIBROSIS COMPARED TO 10 MUG/ML PTX. THESE DATA SUGGEST PTX HAS DIFFERENT BIOLOGICAL EFFECT ON RWA 264.7 CELLS DEPENDING ON THE CONCENTRATION OF 10 MUG/ML AND 300 MUG/ML PTX. THE LOW DOSE 10 MUG/ML PTX ENHANCED RAS/NFKB SIGNALING, PROLIFERATION, DIFFERENTIATION, AND INFLAMMATION, PARTICULARLY, IT STIMULATED NEUROMUSCULAR AND OSTEOBLASTIC DIFFERENTIATION, INNATE IMMUNITY, AND CELL-MEDIATED IMMUNITY, BUT ATTENUATED ER STRESS, FIBROSIS, ANGIOGENESIS, AND CHRONIC INFLAMMATION, WHILE THE HIGH DOSE 300 MUG/ML PTX WAS FOUND TO ALLEVIATE THE 10 MUG/ML PTX-INDUCED BIOLOGICAL EFFECTS, RESULTED IN THE SUPPRESSION OF RAS/NFKB SIGNALING, PROLIFERATION, NEUROMUSCULAR AND OSTEOBLASTIC DIFFERENTIATION, AND INFLAMMATION.	2022	

2 6586  32 TUBASTATIN, A SELECTIVE HISTONE DEACETYLASE 6 INHIBITOR SHOWS ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS. EPIGENETIC MODIFICATIONS REPRESENT A PROMISING NEW APPROACH TO MODULATE CELL FUNCTIONS AS OBSERVED IN AUTOIMMUNE DISEASES. EMERGING EVIDENCE SUGGESTS THE UTILITY OF HDAC INHIBITORS IN THE TREATMENT OF CHRONIC IMMUNE AND INFLAMMATORY DISORDERS. HOWEVER, CLASS AND ISOFORM SELECTIVE INHIBITION OF HDAC IS CURRENTLY FAVORED AS IT LIMITS THE TOXICITY THAT HAS BEEN OBSERVED WITH PAN-HDAC INHIBITORS. HDAC6, A MEMBER OF THE HDAC FAMILY, WHOSE MAJOR SUBSTRATE IS ALPHA-TUBULIN, IS BEING INCREASINGLY IMPLICATED IN THE PATHOGENESIS OF INFLAMMATORY DISORDERS. THE PRESENT STUDY WAS CARRIED OUT TO STUDY THE POTENTIAL ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS OF HDAC6 SELECTIVE INHIBITOR TUBASTATIN. TUBASTATIN, A POTENT HUMAN HDAC6 INHIBITOR WITH AN IC50 OF 11 NM SHOWED SIGNIFICANT INHIBITION OF TNF-ALPHA AND IL-6 IN LPS STIMULATED HUMAN THP-1 MACROPHAGES WITH AN IC50 OF 272 NM AND 712 NM RESPECTIVELY. ADDITIONALLY, TUBASTATIN INHIBITED NITRIC OXIDE (NO) SECRETION IN MURINE RAW 264.7 MACROPHAGES DOSE DEPENDENTLY WITH AN IC50 OF 4.2 MUM AND INDUCED ALPHA-TUBULIN HYPERACETYLATION CORRESPONDING TO HDAC6 INHIBITION IN THP-1 CELLS WITHOUT AFFECTING THE CELL VIABILITY. TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF PAW VOLUME AT 30 MG/KG I.P. IN A FREUND'S COMPLETE ADJUVANT (FCA) INDUCED ANIMAL MODEL OF INFLAMMATION. THE DISEASE MODIFYING ACTIVITY OF TUBASTATIN WAS ALSO EVIDENT IN COLLAGEN INDUCED ARTHRITIS DBA1 MOUSE MODEL AT 30 MG/KG I.P. THE SIGNIFICANT ATTENUATION OF CLINICAL SCORES (~70%) BY TUBASTATIN WAS CONFIRMED HISTOPATHOLOGICALLY AND WAS FOUND COMPARABLE TO DEXAMETHASONE (~90% INHIBITION OF CLINICAL SCORES). TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF IL-6 IN PAW TISSUES OF ARTHRITIC MICE. THE PRESENT WORK HAS DEMONSTRATED ANTI-INFLAMMATORY AND ANTIRHEUMATIC EFFECTS OF A SELECTIVE HDAC6 INHIBITOR TUBASTATIN.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
3  916  35 CHRONIC HIGH GLUCOSE AND INSULIN STIMULATE BONE-MARROW STROMAL CELLS ADIPOGENIC DIFFERENTIATION IN YOUNG SPONTANEOUSLY HYPERTENSIVE RATS. WE EVALUATED WHETHER GENETIC PREDISPOSITION IS SUFFICIENT TO INDUCE CHANGES DUE TO CHRONIC HIGH GLUCOSE (HG; 25 MMOL/L) IN THE PRESENCE OR ABSENCE OF INSULIN (HGI; 10 MUG/ML) ON OSTEOGENIC DIFFERENTIATION AND MARKERS IN BONE-MARROW MESENCHYMAL STEM CELLS (BMSCS) FROM YOUNG WISTAR (WBMSCS) AND SPONTANEOUS HYPERTENSIVE RATS (SBMSCS) WITHOUT HYPERTENSION. HG SUPPRESSED OSTEOGENIC DIFFERENTIATION IN BOTH THE STRAINS, OBSERVED BY MINERALIZATION INHIBITION AND DECREASED LEVELS OF THE OSTEOGENIC MARKERS RUNX2, OSTERIX, OSTEOPONTIN, AND BONE SIALOPROTEIN, COMPARED TO OSTEOGENIC MEDIUM (OM) CELLS. IN WBMSCS, THE EFFECTS OF HG WERE ASSOCIATED WITH THE DOWN REGULATION OF ERK1/2 AND UP REGULATION OF P38 ACTIVITIES; HOWEVER, HGI DID NOT REVERT THE EFFECTS OF HG ON MAPK ACTIVITIES. MOREOVER, HG DID NOT AFFECT MAPK SIGNALING IN SBMSCS COMPARED TO THAT IN OM. HGI INCREASED MINERALIZATION IN WBMSCS COMPARED TO THAT IN OM, BUT NOT IN SBMSCS. HIGH EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AND GLUCOSE TRANSPORTER TYPE 4 IN OM COULD BE RELATED WITH THE PREDISPOSITION TO ADIPOGENIC DIFFERENTIATION NOTED IN SBMSCS AND WAS CONFIRMED BY EMERGENCE OF ADIPOCYTE-LIKE CELLS BY HGI TREATMENT. DOWNREGULATION OF P38 AND UPREGULATION OF JNK ACTIVITIES WERE OBSERVED IN BOTH BMSCS TREATED WITH HGI COMPARED TO THOSE TREATED BY HG. MA (OSMOTIC CONTROL) ALSO SUPPRESSED OSTEOGENIC DIFFERENTIATION IN BOTH THE STRAINS. IN CONCLUSION, WE DEMONSTRATED THAT SBMSCS FROM YOUNG SPONTANEOUS HYPERTENSIVE RATS, WITHOUT HYPERTENSION BUT WITH GENETIC AND EPIGENETIC PREDISPOSITION, EXHIBITED DECREASED OSTEOBLASTIC DIFFERENTIATION UNDER HG AND HGI DID NOT REVERT THE EFFECTS OF HG IN SBMSCS BUT INCREASED ADIPOGENIC DIFFERENTIATION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
4  428  22 ANTI-INFLAMMATORY ACTIVITY OF MIODESIN: MODULATION OF INFLAMMATORY MARKERS AND EPIGENETIC EVIDENCE. PURPOSE: TO INVESTIGATE THE EFFECTS OF A COMBINED HERBAL MEDICINE MIODESIN ON THE INFLAMMATORY RESPONSE OF KEY CELLS INVOLVED IN THE ACUTE AND CHRONIC INFLAMMATORY PROCESSES AS WELL AS THE POSSIBLE EPIGENETIC INVOLVEMENT. METHODS: AFTER THE ESTABLISHMENT OF THE IC(50) DOSE, THE CHONDROCYTE, KERATINOCYTE, AND MACROPHAGE CELL LINES WERE PRETREATED FOR 2 HOURS WITH MIODESIN (200 MUG/ML) AND STIMULATED WITH LPS (1 MUG/ML) FOR 24 HOURS. THE SUPERNATANT WAS USED TO MEASURE THE LEVELS OF CYTOKINES (IL-1BETA, IL-6, IL-8, AND TNF-ALPHA) AND CHEMOKINES (CCL2, CCL3, AND CCL5), AND THE CELLS WERE USED TO EXTRACT THE MRNA FOR THE TRANSCRIPTION FACTOR (NF-KAPPABETA), INFLAMMATORY ENZYMES (COX-1, COX-2, PLA2, AND INOS), AND CHEMOKINES (CCL2, CCL3, AND CCL5). RESULTS: MIODESIN INHIBITED THE RELEASE OF LPS-INDUCED CYTOKINES (IL-1BETA, IL-6, IL-8, AND TNF-ALPHA; P < 0.01) AND CHEMOKINES (CCL2, CCL3, AND CCL5; P < 0.01) AND THE EXPRESSION OF THE TRANSCRIPTION FACTOR (NF-KAPPABETA; P < 0.01), INFLAMMATORY ENZYMES (COX-1, COX-2, PLA2, INOS; P < 0.01), AND CHEMOKINES (CCL2, CCL3, AND CCL5; P < 0.01). IN ADDITION, THE EVALUATION OF EPIGENETIC MECHANISM REVEALED THAT MIODESIN DID NOT INDUCE CHANGES IN DNA METHYLATION, ASSURING THE GENETIC SAFENESS OF THE COMPOUND IN TERMS OF THE INFLAMMATORY RESPONSE. CONCLUSIONS: MIODESIN PRESENTS ANTI-INFLAMMATORY PROPERTIES, INHIBITING HYPERACTIVATION OF CHONDROCYTES, KERATINOCYTES, AND MACROPHAGES, INVOLVING EPIGENETICS IN SUCH EFFECTS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
5 3526  37 IL-6 AND SIL-6R INDUCES STAT3-DEPENDENT DIFFERENTIATION OF HUMAN VSMCS INTO OSTEOBLAST-LIKE CELLS THROUGH JMJD2B-MEDIATED HISTONE DEMETHYLATION OF RUNX2. INFLAMMATION AND VASCULAR CALCIFICATION ARE INDEPENDENT RISK FACTORS OF CARDIOVASCULAR EVENTS. VASCULAR SMOOTH MUSCLE CELLS (VSMCS) EXHIBIT OSTEOBLAST-LIKE CHARACTERISTICS IN RESPONSE TO VARIOUS STIMULI SUCH AS OXIDIZED CHOLESTEROL AND INFLAMMATION. HOWEVER THE PRECISE MECHANISM OF TRANSCRIPTIONAL REGULATION OF VSMCS BY INFLAMMATORY STIMULI REMAINS UNCLEAR. WE INVESTIGATED THE PROCESS AND MECHANISMS OF INFLAMMATORY CYTOKINE-INDUCED TRANSFORMATION OF HUMAN VSMCS (HVSMCS) INTO OSTEOBLAST-LIKE CELLS, WITH A SPECIAL FOCUS ON EPIGENETIC CHANGES. OUR RESULTS DEMONSTRATED: (1) INTERLEUKIN-6 (IL-6)/SOLUBLE INTERLEUKIN-6 RECEPTOR (SIL-6R) INDUCED TRANSFORMATION OF HVSMCS INTO AN OSTEOBLAST PHENOTYPE, WITH SUBSEQUENT VASCULAR CALCIFICATION, BASED ON THE RESULTS OF ALIZARIN RED S STAINING AND O-CRESOLPHTHALEIN COMPLEXONE METHOD; (2) IL-6/SIL-6R ACCELERATED THE EXPRESSION OF RUNT-RELATED TRANSCRIPTION FACTOR 2 (RUNX2) BASED ON THE RESULTS OF QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION; (3) KNOCKDOWN OF SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION (STAT) 3 REDUCED IL-6/SIL-6R-INDUCED RUNX2 MRNA EXPRESSION AND OSTEOBLAST TRANSDIFFERENTIATION OF HVSMCS; (4) CHROMATIN IMMUNOPRECIPITATION (CHIP) COUPLED WITH PCR (CHIP-PCR) IDENTIFIED A STAT-BINDING SITE IN RUNX2 PROMOTER REGION CONTAINING TRIMETHYLATED HISTONE 3 LYSINE 9 (H3K9ME3), A TRANSCRIPTIONAL REPRESSOR, AND H3K4ME3, A TRANSCRIPTIONAL ENHANCER. STIMULATION WITH IL-6/SIL-6R SUPPRESSED H3K9ME3 BUT NOT H3K4ME3 THROUGH THE RECRUITMENT OF JUMONJI DOMAIN-CONTAINING PROTEIN (JMJD) 2B, A HISTONE LYSINE DEMETHYLASE, AT THE STAT-BINDING SITE IN RUNX2 PROMOTER REGION; (5) IL-6/SIL-6R-INDUCED RUNX2 GENE EXPRESSION WAS INHIBITED IN HVSMCS PRETREATED WITH JIB04, JMJD2 INHIBITOR, AND THE INHIBITORY EFFECT WAS JIB04 DOSE-DEPENDENT. OUR RESULTS INDICATE THAT THE IL-6/STAT3/JMJD2B PATHWAY REGULATES HVSMCS DIFFERENTIATION INTO OSTEOBLAST-LIKE CELLS, WHICH SUGGEST ITS PATHOGENIC ROLE IN VASCULAR CALCIFICATION ASSOCIATED WITH CHRONIC INFLAMMATION.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
6 2032  29 EPIGENETIC CHANGES IN P21 EXPRESSION IN RENAL CELLS AFTER EXPOSURE TO BROMATE. THIS STUDY TESTED THE HYPOTHESIS THAT BROMATE (KBRO3)-INDUCED RENAL CELL DEATH IS MEDIATED BY EPIGENETIC MECHANISMS. GLOBAL DNA METHYLATION, AS ASSESSED BY 5-METHYLCYTOSINE STAINING, WAS NOT CHANGED IN NORMAL RAT KIDNEY CELLS TREATED WITH ACUTE CYTOTOXIC DOSES OF KBRO3 (100 AND 200 PPM), AS COMPARED WITH CONTROLS. HOWEVER, KBRO3 TREATMENT DID INCREASE P38, P53 AND HISTONE 2AX (H2AX) PHOSPHORYLATION, AND P21 EXPRESSION. TREATMENT OF CELLS WITH INHIBITORS OF DNA METHYLTRANSFERASE (5-AZACYTIDINE OR 5-AZA) AND HISTONE DEACETYLASE (TRICHOSTATIN A OR TSA) IN ADDITION TO KBRO3 INCREASED CYTOTOXICITY, AS COMPARED WITH CELLS EXPOSED TO KBRO3 ALONE. 5-AZA AND TSA CO-TREATMENT DID NOT ALTER P38 OR P53 PHOSPHORYLATION, BUT SLIGHTLY DECREASED H2AX PHOSPHORYLATION AND SIGNIFICANTLY DECREASED P21 EXPRESSION. WE ALSO ASSESSED EPIGENETIC CHANGES IN CELLS TREATED UNDER SUB-CHRONIC CONDITIONS WITH ENVIRONMENTALLY RELEVANT CONCENTRATIONS OF KBRO3. UNDER THESE CONDITIONS (0-10PPM KBRO3 FOR UP TO 18 DAYS), WE DETECTED NO INCREASES IN CELL DEATH OR DNA DAMAGE. IN CONTRAST, SLIGHT ALTERATIONS WERE DETECTED IN THE PHOSPHORYLATION OF H2AX, P38, AND P53. SUB-CHRONIC LOW-DOSE KBRO3 TREATMENT ALSO INDUCED A BIPHASIC RESPONSE IN P21 EXPRESSION, WITH LOWER CONCENTRATIONS INCREASING EXPRESSION, BUT HIGHER CONCENTRATIONS DECREASING EXPRESSION. METHYLATION-SPECIFIC PCR DEMONSTRATED THAT SUB-CHRONIC KBRO3 TREATMENT ALTERED THE METHYLATION OF CYTOSINE BASES IN THE P21 GENE, AS COMPARED WITH CONTROLS, CORRELATING TO ALTERATIONS IN P21 PROTEIN EXPRESSION. COLLECTIVELY, THESE DATA SHOW THE NOVEL FINDING THAT KBRO3-INDUCED RENAL CELL DEATH IS ALTERED BY INHIBITORS OF EPIGENETIC MODIFYING ENZYMES AND THAT KBRO3 ITSELF INDUCES EPIGENETIC CHANGES IN THE P21 GENE.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
7 1658  29 DOWN-REGULATION OF A PRO-APOPTOTIC PATHWAY REGULATED BY PCAF/ADA3 IN EARLY STAGE GASTRIC CANCER. THE LOSS OF P300/CBP-ASSOCIATED PROTEIN (PCAF) EXPRESSION IS ASSOCIATED WITH POOR CLINICAL OUTCOME IN GASTRIC CANCER, AND A POTENTIAL BIO-MARKER FOR INVASIVE AND AGGRESSIVE TUMORS. HOWEVER, THE MECHANISM LINKING LOSS OF PCAF TO THE ONSET OF GASTRIC CANCER HAS NOT BEEN IDENTIFIED. GIVEN THAT PCAF AND ITS BINDING PARTNER TRANSCRIPTIONAL ADAPTOR PROTEIN 3 (ADA3) WERE RECENTLY SHOWN TO REGULATE THE INTRINSIC (MITOCHONDRIAL) PATHWAY TO APOPTOSIS VIA EPIGENETIC REGULATION OF PHOSPHOFURIN ACIDIC CLUSTER SORTING PROTEINS 1 AND 2 (PACS1, PACS2), WE ANALYZED PCAF, ADA3, AND PACS1/2 EXPRESSION IN 99 PATIENT-MATCHED SURGICAL SAMPLES RANGING FROM NORMAL GASTRIC MUCOSA, THROUGH PRE-MALIGNANT CHRONIC GASTRITIS AND INTESTINAL METAPLASIA TO STAGE I-III INVASIVE CANCERS. PCAF MRNA LEVELS WERE NOT REDUCED IN EITHER PRE-MALIGNANT STATE BUT WERE SIGNIFICANTLY DOWN-REGULATED IN ALL STAGES OF GASTRIC CANCER, COMMENCING AT AJCC STAGE I (P < 0.05), THUS LINKING REDUCED PCAF EXPRESSION WITH EARLY MALIGNANT CHANGE. FURTHERMORE, PATIENTS WITH COMBINED REDUCTION OF PCAF AND PACS1 HAD SIGNIFICANTLY POORER OVERALL SURVIVAL (P = 0.0257), CONFIRMED IN AN INDEPENDENT DATASET OF 359 PATIENTS (P = 5.8 X 10E-6). AT THE PROTEIN LEVEL, PCAF, ADA3, AND PACS1 EXPRESSION WERE ALL SIGNIFICANTLY DOWN-REGULATED IN INTESTINAL-TYPE GASTRIC CANCER, AND CORRELATED WITH REDUCED PROGRESSION FREE SURVIVAL. WE CONCLUDE THAT A PRO-APOPTOTIC MECHANISM CENTERED ON THE INTRINSIC (MITOCHONDRIAL) PATHWAY AND REGULATED BY PCAF/ADA3 CAN INFLUENCE THE PROGRESSION FROM PREMALIGNANT TO MALIGNANT CHANGE, AND THUS ACT AS A TUMOR SUPPRESSION MECHANISM IN GASTRIC CANCER.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
8 4001  39 LOSS OF MEN1 LEADS TO RENAL FIBROSIS AND DECREASES HGF-ADAMTS5 PATHWAY ACTIVITY VIA AN EPIGENETIC MECHANISM. BACKGROUND: RENAL FIBROSIS IS A SERIOUS CONDITION THAT RESULTS IN THE DEVELOPMENT OF CHRONIC KIDNEY DISEASES. THE MEN1 GENE IS AN EPIGENETIC REGULATOR THAT ENCODES THE MENIN PROTEIN AND ITS ROLE IN KIDNEY TISSUE REMAINS UNCLEAR. METHODS: KIDNEY HISTOLOGY WAS EXAMINED ON PARAFFIN SECTIONS STAINED WITH HEMATOXYLIN-EOSIN STAINING. MASSON'S TRICHROME STAINING AND SIRIUS RED STAINING WERE USED TO ANALYZE RENAL FIBROSIS. GENE AND PROTEIN EXPRESSION WERE DETERMINED BY QUANTITATIVE REAL-TIME PCR (QPCR) AND WESTERN BLOT, RESPECTIVELY. IMMUNOHISTOCHEMISTRY STAINING IN THE KIDNEY TISSUES FROM MICE OR PATIENTS WAS USED TO EVALUATE PROTEIN LEVELS. FLOW CYTOMETRY WAS USED TO ANALYZE THE CELL CYCLE DISTRIBUTIONS AND APOPTOSIS. RNA-SEQUENCING WAS PERFORMED FOR DIFFERENTIAL EXPRESSION GENES IN THE KIDNEY TISSUES OF THE MEN1F/F AND MEN1?/? MICE. CHROMATIN IMMUNOPRECIPITATION SEQUENCING (CHIP-SEQ) WAS CARRIED OUT FOR IDENTIFICATION OF MENIN- AND H3K4ME3-ENRICHED REGIONS WITHIN THE WHOLE GENOME IN THE MOUSE KIDNEY TISSUE. CHIP-QPCR ASSAYS WERE PERFORMED FOR OCCUPANCY OF MENIN AND H3K4ME3 AT THE GENE PROMOTER REGIONS. LUCIFERASE REPORTER ASSAY WAS USED TO DETECT THE PROMOTER ACTIVITY. THE EXACERBATED UNILATERAL URETERAL OBSTRUCTION (UUO) MODELS IN THE MEN1F/F AND MEN1?/? MICE WERE USED TO ASSESS THE PHARMACOLOGICAL EFFECTS OF RH-HGF ON RENAL FIBROSIS. RESULTS: THE EXPRESSION OF MEN1 IS REDUCE IN KIDNEY TISSUES OF FIBROTIC MOUSE AND HUMAN DIABETIC PATIENTS AND TREATMENT WITH FIBROTIC FACTOR RESULTS IN THE DOWNREGULATION OF MEN1 EXPRESSION IN RENAL TUBULAR EPITHELIAL CELLS (RTECS). DISRUPTION OF MEN1 IN RTECS LEADS TO HIGH EXPRESSION OF ALPHA-SMA AND COLLAGEN 1, WHEREAS MEN1 OVEREXPRESSION RESTRAINS EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) INDUCED BY TGF-BETA TREATMENT. CONDITIONAL KNOCKOUT OF MEN1 RESULTED IN CHRONIC RENAL FIBROSIS AND UUO-INDUCED TUBULOINTERSTITIAL FIBROSIS (TIF), WHICH IS ASSOCIATED WITH AN INCREASED INDUCTION OF EMT, G2/M ARREST AND JNK SIGNALING. MECHANISTICALLY, MENIN RECRUITS AND INCREASES H3K4ME3 AT THE PROMOTER REGIONS OF HEPATOCYTE GROWTH FACTOR (HGF) AND A DISINTEGRIN AND METALLOPROTEINASE WITH THROMBOSPONDIN MOTIFS 5 (ADAMTS5) GENES AND ENHANCES THEIR TRANSCRIPTIONAL ACTIVATION. IN THE UUO MICE MODEL, EXOGENOUS HGF RESTORED THE EXPRESSION OF ADAMTS5 AND AMELIORATED RENAL FIBROSIS INDUCED BY MEN1 DEFICIENCY. CONCLUSIONS: THESE FINDINGS DEMONSTRATE THAT MEN1 IS AN ESSENTIAL ANTIFIBROTIC FACTOR IN RENAL FIBROGENESIS AND COULD BE A POTENTIAL TARGET FOR ANTIFIBROTIC THERAPY.	2022	
                                
9 3128  37 GIPC-REGULATED IGFBP-3 PROMOTES HSC MIGRATION IN VITRO AND PORTAL HYPERTENSION IN VIVO THROUGH A BETA1-INTEGRIN PATHWAY. BACKGROUND & AIMS: TRANSFORMING GROWTH FACTOR (TGF-BETA)-INDUCED ACTIVATION OF QUIESCENT HEPATIC STELLATE CELLS (HSCS) AND THEIR TRANSFORMATION TO MYOFIBROBLASTS IS A KEY EVENT IN LIVER FIBROSIS AND PORTAL HYPERTENSION. GIPC (ALSO REFERRED TO AS SYNECTIN) IS A DOWNSTREAM SIGNAL ACTIVATION MOLECULE OF TGF-BETA AND OTHER RECEPTORS. IN THIS STUDY, WE SOUGHT TO IDENTIFY NOVEL GENES TARGETED BY TGF-BETA AND GIPC AND ELUCIDATE IF AND HOW THEY MAY CONTRIBUTE TO LIVER FIBROSIS. METHODS: WE PERFORMED SEQUENTIAL MESSENGER RNA SEQUENCING ANALYSIS ON TGF-BETA-STIMULATED HSCS AND THEN ON TGF-BETA-STIMULATED HSCS IN THE PRESENCE AND ABSENCE OF GIPC ALSO REFERRED TO AS SYNECTIN (GIPC) KNOCKDOWN. INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN-3 (IGFBP-3) TRANSPORT PROTEIN EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC. QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, TARGETED CHROMATIN IMMUNOPRECIPITATION, AND WESTERN BLOT ANALYSIS WERE DONE FOR FURTHER CONFIRMATION. RESULTS: IGFBP-3, AN INSULIN GROWTH FACTOR TRANSPORT PROTEIN, EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC, WHICH WAS CONFIRMED BY QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, AND WESTERN BLOT ANALYSIS. TARGETED CHROMATIN IMMUNOPRECIPITATION SHOWED THAT GIPC INCREASES THE HISTONE 3 LYSINE 27 (H3K27) ACETYLATION ACTIVATING MARK AND CONCURRENTLY DECREASES THE H3K27 INHIBITORY TRIMETHYLATION (H3K27M3) MARK, PROVIDING AN EPIGENETIC CORRELATE TO THE GENE REGULATION CHANGES. IN VIVO, GLOBAL KNOCKOUT OF IGFBP-3 MICE RESULTED IN ATTENUATION OF HSC ACTIVATION MARKERS AND ATTENUATION OF PORTAL PRESSURE IN RESPONSE TO CHRONIC LIVER INJURY MODELS. ANALYSIS OF SERUM LEVELS FROM CIRRHOTIC PATIENTS ALSO SHOWED AN IGFBP-3 INCREASE OF MORE THAN 2-FOLD COMPARED WITH HEALTHY CONTROLS. FINALLY, IN VITRO MECHANISM STUDIES SHOWED THAT IGFBP-3 PROMOTES HSC MIGRATION THROUGH INTEGRIN-DEPENDENT PHOSPHORYLATION OF PROTEIN KINASE B. CONCLUSIONS: TGF-BETA UP-REGULATES IGFBP-3 THROUGH GIPC, LEADING TO INCREASED HSC MIGRATION IN VITRO AND PROMOTES PORTAL HYPERTENSION IN VIVO. THESE STUDIES SUPPORT THE ROLE OF IGFBP-3 AS A POTENTIAL PATHOPHYSIOLOGIC TARGET OR BIOMARKER IN CHRONIC LIVER DISEASE.	2020	
                                                                                                                                                                                                                                                                                                                        
10 1966  34 EPIGENETIC ALTERATION OF PRKCDBP IN COLORECTAL CANCERS AND ITS IMPLICATION IN TUMOR CELL RESISTANCE TO TNFALPHA-INDUCED APOPTOSIS. PURPOSE: PRKCDBP IS A PUTATIVE TUMOR SUPPRESSOR IN WHICH ALTERATION HAS BEEN OBSERVED IN SEVERAL HUMAN CANCERS. WE INVESTIGATED EXPRESSION AND FUNCTION OF PRKCDBP IN COLORECTAL CELLS AND TISSUES TO EXPLORE ITS CANDIDACY AS A SUPPRESSOR IN COLORECTAL TUMORIGENESIS. EXPERIMENTAL DESIGN: EXPRESSION AND METHYLATION STATUS OF PRKCDBP AND ITS EFFECT ON TUMOR GROWTH WERE EVALUATED. TRANSCRIPTIONAL REGULATION BY NF-KAPPAB SIGNALING WAS DEFINED BY LUCIFERASE REPORTER AND CHROMATIN IMMUNOPRECIPITATION ASSAYS. RESULTS: PRKCDBP EXPRESSION WAS HARDLY DETECTABLE IN 29 OF 80 (36%) PRIMARY TUMORS AND 11 OF 19 (58%) CELL LINES, AND ITS ALTERATION CORRELATED WITH TUMOR STAGE AND GRADE. PROMOTER HYPERMETHYLATION WAS COMMONLY FOUND IN CANCERS. PRKCDBP EXPRESSION INDUCED THE G(1) CELL-CYCLE ARREST AND INCREASED CELLULAR SENSITIVITY TO VARIOUS APOPTOTIC STRESSES. PRKCDBP WAS INDUCED BY TNFALPHA, AND ITS LEVEL CORRELATED WITH TUMOR CELL SENSITIVITY TO TNFALPHA-INDUCED APOPTOSIS. PRKCDBP INDUCTION BY TNFALPHA WAS DISRUPTED BY BLOCKING NF-KAPPAB SIGNALING WHILE IT WAS ENHANCED BY RELA TRANSFECTION. THE PRKCDBP PROMOTER ACTIVITY WAS INCREASED IN RESPONSE TO TNFALPHA, AND THIS RESPONSE WAS ABOLISHED BY DISRUPTION OF A KAPPAB SITE IN THE PROMOTER. PRKCDBP DELAYED THE FORMATION AND GROWTH OF XENOGRAFT TUMORS AND IMPROVED TUMOR RESPONSE TO TNFALPHA-INDUCED APOPTOSIS. CONCLUSIONS: PRKCDBP IS A PROAPOPTOTIC TUMOR SUPPRESSOR WHICH IS COMMONLY ALTERED IN COLORECTAL CANCER BY PROMOTER HYPERMETHYLATION, AND ITS GENE TRANSCRIPTION IS DIRECTLY ACTIVATED BY NF-KAPPAB IN RESPONSE TO TNFALPHA. THIS SUGGESTS THAT PRKCDBP INACTIVATION MAY CONTRIBUTE TO TUMOR PROGRESSION BY REDUCING CELLULAR SENSITIVITY TO TNFALPHA AND OTHER STRESSES, PARTICULARLY UNDER CHRONIC INFLAMMATORY MICROENVIRONMENT.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
11 2896  26 GASTRIC ENTEROCHROMAFFIN-LIKE CELL HYPERPLASIA AND NEOPLASIA IN THE RAT: AN INDIRECT EFFECT OF THE HISTAMINE H2-RECEPTOR ANTAGONIST, BL-6341. ORAL ADMINISTRATION OF BL-6341 HYDROCHLORIDE, A LONG-ACTING HISTAMINE H2-RECEPTOR ANTAGONIST, TO RATS FOR 2 YEARS AT DOSES OF 10, 55 OR 300 MG/KG/DAY RESULTED IN SEVERAL CHANGES IN THE FUNDIC (OXYNTIC) MUCOSA OF THE GLANDULAR STOMACH. THE MOST SIGNIFICANT ALTERATION WAS A PROLIFERATION OF ARGYROPHIL ENDOCRINE CELLS THAT WAS DEMONSTRATED TO BE ENTEROCHROMAFFIN-LIKE (ECL) CELLS. THE ECL CELL PROLIFERATION CONSISTED OF A CONTINUUM OF CHANGES INVOLVING DIFFUSE HYPERPLASIA, FOCAL ADENOMATOUS HYPERPLASIA, AND CARCINOID TUMOR FORMATION AT THE HIGHEST DOSE LEVEL OF 300 MG/KG. AT 55 MG/KG ONLY ECL CELL HYPERPLASIA OCCURRED, AND AT THE LOW DOSE OF 10 MG/KG THERE WERE NO REMARKABLE PROLIFERATIVE CHANGES. THE REFERENCE COMPOUND, CIMETIDINE (950 MG/KG), PRODUCED A DEGREE OF ECL CELL PROLIFERATION THAT WAS SLIGHTLY LESS, BUT NOT SIGNIFICANTLY DIFFERENT THAN, THAT OBSERVED WITH 55 MG/KG OF BL-6341. DOSE-RELATED ELEVATIONS OF SERUM GASTRIN WERE OBSERVED WITH BL-6341, WHILE CIMETIDINE PRODUCED HYPERGASTRINEMIA THAT WAS GENERALLY INTERMEDIATE BETWEEN THAT PRODUCED BY THE MIDDLE AND LOW DOSES OF BL-6341. THE HYPERGASTRINEMIA RESULTED FROM THE PHARMACOLOGIC INHIBITION OF ACID SECRETION, WHICH IS THE NEGATIVE FEEDBACK MECHANISM CONTROLLING THE PRODUCTION OF GASTRIN. ONLY THE 300 MG/KG DOSE OF BL-6341 PRODUCED A SIGNIFICANT, SUSTAINED (24 HOURS) HYPERGASTRINEMIA AND CARCINOID TUMORS. THE CHRONIC, SUSTAINED HYPERGASTRINEMIA WAS CONSIDERED TO BE THE PRIMARY CAUSE OF THE ECL CELL CARCINOID NEOPLASIA. ALL GENETIC TOXICOLOGY TESTS PERFORMED WITH BL-6341 WERE NEGATIVE. IT WAS CONCLUDED THAT THE DEMONSTRATED HYPERGASTRINEMIA REPRESENTS AN INDIRECT, HORMONAL, EPIGENETIC MECHANISM OF TUMORIGENESIS.	1988	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
12 6294  17 THE PROINFLAMMATORY CYTOKINE TNFALPHA INDUCES DNA DEMETHYLATION-DEPENDENT AND -INDEPENDENT ACTIVATION OF INTERLEUKIN-32 EXPRESSION. IL-32 IS A CYTOKINE INVOLVED IN PROINFLAMMATORY IMMUNE RESPONSES TO BACTERIAL AND VIRAL INFECTIONS. HOWEVER, THE ROLE OF EPIGENETIC EVENTS IN THE REGULATION OF IL-32 GENE EXPRESSION IS UNDERSTUDIED. HERE WE SHOW THAT IL-32 IS REPRESSED BY DNA METHYLATION IN HEK293 CELLS. USING CHIP SEQUENCING, LOCUS-SPECIFIC METHYLATION ANALYSIS, CRISPR/CAS9-MEDIATED GENOME EDITING, AND RT-QPCR (QUANTITATIVE RT-PCR) AND IMMUNOBLOT ASSAYS, WE FOUND THAT SHORT-TERM TREATMENT (A FEW HOURS) WITH THE PROINFLAMMATORY CYTOKINE TUMOR NECROSIS FACTOR ALPHA (TNFALPHA) ACTIVATES IL-32 IN A DNA DEMETHYLATION-INDEPENDENT MANNER. IN CONTRAST, PROLONGED TNFALPHA TREATMENT (SEVERAL DAYS) INDUCED DNA DEMETHYLATION AT THE PROMOTER AND A CPG ISLAND IN THE IL-32 GENE IN A TET (TEN-ELEVEN TRANSLOCATION) FAMILY ENZYME- AND NF-KAPPAB-DEPENDENT MANNER. NOTABLY, THE HYPOMETHYLATION STATUS OF TRANSCRIPTIONAL REGULATORY ELEMENTS IN IL-32 WAS MAINTAINED FOR A LONG TIME (SEVERAL WEEKS), CAUSING ELEVATED IL-32 EXPRESSION EVEN IN THE ABSENCE OF TNFALPHA. CONSIDERING THAT IL-32 CAN, IN TURN, INDUCE TNFALPHA EXPRESSION, WE SPECULATE THAT SUCH FEEDFORWARD EVENTS MAY CONTRIBUTE TO THE TRANSITION FROM AN ACUTE INFLAMMATORY RESPONSE TO CHRONIC INFLAMMATION.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
13 6148  33 THE EXPRESSION OF TRANSCRIPTION FACTORS MECP2 AND CREB IS MODULATED IN INFLAMMATORY PELVIC PAIN. EARLY ACTIVATION OF TRANSCRIPTION FACTORS IS ONE OF THE EPIGENETIC MECHANISMS CONTRIBUTING TO THE INDUCTION AND MAINTENANCE OF CHRONIC PAIN STATES. PREVIOUS STUDIES IDENTIFIED THE CHANGES IN A NUMBER OF NOCICEPTION-RELATED GENES, SUCH AS CALCITONIN GENE-RELATED PEPTIDE (CGRP), SUBSTANCE P (SP), AND BRAIN-DERIVED NEUROTROPIC FACTOR (BDNF) IN THE PELVIC ORGANS AFTER TRANSIENT COLONIC INFLAMMATION. THE GENE AND PROTEIN EXPRESSION OF THESE NEUROPEPTIDES COULD BE MODULATED BY TRANSCRIPTION FACTORS METHYL-CPG-BINDING PROTEIN 2 (MECP2) AND CAMP RESPONSE ELEMENT-BINDING PROTEIN (CREB). IN THIS STUDY, WE AIMED TO EVALUATE TIME-DEPENDENT CHANGES IN THE EXPRESSION LEVELS OF MECP2 AND CREB IN THE LUMBOSACRAL (LS) SPINAL CORD AND SENSORY GANGLIA AFTER INFLAMMATION-INDUCED PELVIC PAIN IN RAT. ADULT SPRAGUE-DAWLEY RATS WERE TREATED WITH 2,4,6-TRINITROBENZENESULFONIC ACID (TNBS) TO INDUCE TRANSIENT COLONIC INFLAMMATION. LS (L6-S2) SPINAL CORD SEGMENTS AND RESPECTIVE DORSAL ROOT GANGLIAS (DRGS) WERE ISOLATED FROM CONTROL AND EXPERIMENTAL ANIMALS AT 1, 2, 6, 24 H AND 3 DAYS POST-TNBS TREATMENT. IMMUNOHISTOCHEMICAL (IHC) LABELING AND WESTERN BLOTTING EXPERIMENTS WERE PERFORMED TO ASSESS THE EXPRESSION OF MECP2, CREB AND THEIR PHOSPHORYLATED FORMS. TOTAL MECP2 EXPRESSION, BUT NOT PHOSPHORYLATED P-MECP2 (PS421MECP2) EXPRESSION WAS DETECTED IN THE CELLS OF THE SPINAL DORSAL HORN UNDER CONTROL CONDITIONS. COLONIC INFLAMMATION TRIGGERED A SIGNIFICANT DECREASE IN THE NUMBER OF MECP2-EXPRESSING NEURONS IN PARALLEL WITH ELEVATED NUMBERS OF PS421MECP2-EXPRESSING CELLS AT 2 H AND 6 H POST-TNBS. THE MAJORITY OF MECP2-POSITIVE CELLS (80 +/- 6%) CO-EXPRESSED CREB. TNBS TREATMENT CAUSED A TRANSIENT UP-REGULATION OF CREB-EXPRESSING CELLS AT 1 H POST-TNBS ONLY. THE NUMBER OF CELLS EXPRESSING PHOSPHORYLATED CREB (PS133CREB) DID NOT CHANGE AT 1 H AND 2 H POST-TNBS, BUT WAS DOWN-REGULATED BY THREE FOLDS AT 6 H POST-TNBS. ANALYSIS OF DRG SECTIONS REVEALED THAT THE NUMBER OF MECP2-POSITIVE NEURONS WAS UP-REGULATED BY TNBS TREATMENT, REACHING THREE-FOLD INCREASE AT 2 H POST-TNBS, AND EIGHT-FOLD INCREASE AT 6 H POST-TNBS (P </= 0.05 TO CONTROL). THESE DATA SHOWED EARLY CHANGES IN MECP2 AND CREB EXPRESSION IN THE DORSAL HORN OF THE SPINAL CORD AND SENSORY GANGLIA AFTER COLONIC INFLAMMATION, SUGGESTING A POSSIBLE CONTRIBUTION MECP2 AND CREB SIGNALING IN THE DEVELOPMENT OF VISCERAL HYPERALGESIA AND PELVIC PAIN FOLLOWING PERIPHERAL INFLAMMATION.	2018	
                                                                                                                          
14 4020  28 LOW-MOLECULAR-WEIGHT FIBROBLAST GROWTH FACTOR 2 ATTENUATES HEPATIC FIBROSIS BY EPIGENETIC DOWN-REGULATION OF DELTA-LIKE1. LIVER FIBROSIS, A MAJOR CAUSE OF END-STAGE LIVER DISEASES, IS CLOSELY REGULATED BY MULTIPLE GROWTH FACTORS AND CYTOKINES. THE CORRELATION OF FIBROBLAST GROWTH FACTOR 2 (FGF2) WITH CHRONIC LIVER INJURY HAS BEEN REPORTED, BUT THE EXACT FUNCTIONS OF DIFFERENT FGF2 ISOFORMS IN LIVER FIBROGENESIS REMAIN UNCLEAR. HERE, WE REPORT ON THE DIFFERENTIAL EXPRESSION PATTERNS AND FUNCTIONS OF LOW- AND HIGH-MOLECULAR-WEIGHT FGF2 (NAMELY, FGF2(LMW) AND FGF2(HMW) , RESPECTIVELY) IN HEPATIC FIBROGENESIS USING A CCL4 -INDUCED MOUSE LIVER FIBROSIS MODEL. FGF2(HMW) DISPLAYED A ROBUST INCREASE IN CCL4 -INDUCED HEPATIC FIBROSIS AND PROMOTED FIBROGENESIS. IN CONTRAST, ENDOGENOUS FGF2(LMW) EXHIBITED A SLIGHT INCREASE IN HEPATIC FIBROSIS AND SUPPRESSED THIS PATHOLOGICAL PROGRESSION. MOREOVER, EXOGENOUS ADMINISTRATION OF RECOMBINANT FGF2(LMW) POTENTLY AMELIORATED CCL4 -INDUCED LIVER FIBROSIS. MECHANISTICALLY, WE SHOWED THAT FGF2(LMW) TREATMENT ATTENUATED HEPATIC STELLATE CELL ACTIVATION AND FIBROSIS BY EPIGENETIC DOWN-REGULATION OF DELTA-LIKE 1 EXPRESSION THROUGH THE P38 MITOGEN-ACTIVATED PROTEIN KINASE PATHWAY. CONCLUSION: FGF2(LMW) AND FGF2(HMW) HAVE DISTINCT ROLES IN LIVER FIBROGENESIS. THESE FINDINGS DEMONSTRATE A POTENT ANTIFIBROTIC EFFECT OF FGF2(LMW) ADMINISTRATION, WHICH MAY PROVIDE A NOVEL APPROACH TO TREAT CHRONIC LIVER DISEASES.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
15 4696  23 NF-KAPPAB REPRESSES RETINOIC ACID RECEPTOR-MEDIATED GPRC5A TRANSACTIVATION IN LUNG EPITHELIAL CELLS TO PROMOTE NEOPLASIA. CHRONIC INFLAMMATION IS ASSOCIATED WITH LUNG TUMORIGENESIS, IN WHICH NF-KAPPAB-MEDIATED EPIGENETIC REGULATION PLAYS A CRITICAL ROLE. LUNG TUMOR SUPPRESSOR G PROTEIN-COUPLED RECEPTOR, FAMILY C, MEMBER 5A (GPRC5A), IS REPRESSED IN MOST NON-SMALL CELL LUNG CANCER (NSCLC); HOWEVER, THE MECHANISMS REMAIN UNCLEAR. HERE, WE SHOW THAT NF-KAPPAB ACTS AS A TRANSCRIPTIONAL REPRESSOR IN SUPPRESSION OF GPRC5A. NF-KAPPAB INDUCED GPRC5A REPRESSION BOTH IN VITRO AND IN VIVO. INTRIGUINGLY, TRANSACTIVATION OF NF-KAPPAB DOWNSTREAM TARGETS WAS NOT REQUIRED, BUT THE TRANSACTIVATION DOMAIN OF RELA/P65 WAS REQUIRED FOR GPRC5A REPRESSION. NF-KAPPAB DID NOT BIND TO ANY POTENTIAL CIS-ELEMENT IN THE GPRC5A PROMOTER. INSTEAD, P65 WAS COMPLEXED WITH RETINOIC ACID RECEPTOR ALPHA/BETA (RARALPHA/BETA) AND RECRUITED TO THE RA RESPONSE ELEMENT SITE AT THE GPRC5A PROMOTER, RESULTING IN DISRUPTED RNA POLYMERASE II COMPLEXING AND SUPPRESSED TRANSCRIPTION. NOTABLY, PHOSPHORYLATION ON SERINE 276 OF P65 WAS REQUIRED FOR INTERACTION WITH RARALPHA/BETA AND REPRESSION OF GPRC5A. MOREOVER, NF-KAPPAB-MEDIATED EPIGENETIC REPRESSION WAS THROUGH SUPPRESSION OF ACETYLATED HISTONE H3K9 (H3K9AC), BUT NOT DNA METHYLATION OF THE CPG ISLANDS, AT THE GPRC5A PROMOTER. CONSISTENTLY, A HISTONE DEACETYLASE INHIBITOR, BUT NOT DNA METHYLATION INHIBITOR, RESTORED GPRC5A EXPRESSION IN NSCLC CELLS. THUS, NF-KAPPAB INDUCES TRANSCRIPTIONAL REPRESSION OF GPRC5A VIA A COMPLEX WITH RARALPHA/BETA AND MEDIATES EPIGENETIC REPRESSION VIA SUPPRESSION OF H3K9AC.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
16 3718  28 INHIBITION OF BCL6B PROMOTES GASTRIC CANCER BY AMPLIFYING INFLAMMATION IN MICE. BACKGROUND: CHRONIC GASTRITIS HAS BEEN DEMONSTRATED TO BE A KEY CAUSE OF GASTRIC CANCER (GC), AND CONTROL OF GASTRIC INFLAMMATION IS REGARDED AS AN EFFECTIVE TREATMENT FOR THE CLINICAL PREVENTION OF GASTRIC CARCINOGENESIS. HOWEVER, THERE REMAINS AN UNMET NEED TO IDENTIFY THE DOMINANT REGULATORS OF GASTRIC ONCOGENESIS-ASSOCIATED INFLAMMATION IN VIVO. METHODS: THE MOUSE MODEL FOR THE STUDY OF INFLAMMATION-ASSOCIATED GC WAS INDUCED BY BENZO[A]PYRENE (BAP) INTRAGASTRIC ADMINISTRATION IN BCL6B(-/-) AND WILDTYPE MICE ON A C57BL/6 BACKGROUND. 5-AZA-2'-DEOXYCYTIDINE (5-AZA), THE DEMETHYLATION DRUG, WAS INTRAPERITONEALLY INJECTED TO RESTORE BCL6B EXPRESSION. HUMAN GC TISSUE ARRAY WAS USED TO ANALYSE PATIENT SURVIVAL BASED ON BCL6B AND CD3 PROTEIN EXPRESSION. RESULTS: BCL6B WAS GRADUALLY DOWNREGULATED BY ITS OWN PROMOTER HYPERMETHYLATION IN PARALLEL TO AN INCREASING INFLAMMATORY RESPONSE DURING THE PROGRESSION OF BAP-INDUCED GASTRIC CARCINOGENESIS IN MICE. MOREOVER, KNOCKOUT OF BCL6B DRAMATICALLY WORSENED THE SEVERITY OF GASTRIC CANCER AND AGGRAVATED THE INFLAMMATORY RESPONSE IN THE BAP-INDUCED MICE GC MODEL. RE-ACTIVATION OF BCL6B BY 5-AZA IMPEDED INFLAMMATORY AMPLIFICATION AND BAP-INDUCED GC DEVELOPMENT, PROLONGING SURVIVAL TIME IN WILDTYPE MICE, WHEREAS NO NOTABLE CURATIVE EFFECT OCCURRED IN BCL6B(-/-) MICE WITH 5-AZA TREATMENT. FINALLY, SIGNIFICANT NEGATIVE CORRELATIONS WERE DETECTED BETWEEN THE MRNA LEVELS OF BCL6B AND INFLAMMATORY CYTOKINES IN HUMAN GC TISSUES; PATIENTS HARBOURING BCL6B-NEGETIVE AND SEVERE-INFLAMMATION GC TUMOURS WERE FOUND TO EXHIBIT THE SHORTEST SURVIVAL TIME. CONCLUSIONS: EPIGENETIC INACTIVATION OF BCL6B PROMOTES GASTRIC CANCER THROUGH AMPLIFICATION OF THE GASTRIC INFLAMMATORY RESPONSE IN VIVO AND OFFERS A NEW APPROACH FOR GC TREATMENT AND REGENERATIVE MEDICINE.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
17 6297  29 THE PROTECTIVE EFFECT OF HBO1 ON CIGARETTE SMOKE EXTRACT-INDUCED APOPTOSIS IN AIRWAY EPITHELIAL CELLS. PURPOSE: EPIGENETIC MODIFICATION IS ONE OF MOST IMPORTANT MECHANISMS UNDERLYING THE PATHOGENESIS OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). THE PURPOSE OF THIS STUDY WAS TO DETERMINE WHETHER HISTONE ACETYLTRANSFERASE BINDING TO ORC1 (HBO1) CAN PROTECT AGAINST CIGARETTE SMOKE (CS)-INDUCED CELL APOPTOSIS AND SUSTAIN NORMAL HISTONE ACETYLATION IN COPD. METHODS: HUMAN LUNG TISSUE SAMPLES WERE OBTAINED FROM PATIENTS WHO UNDERWENT LUNG RESECTION. THE EMPHYSEMA MOUSE MODEL AND HBO1 OVEREXPRESSING MICE WERE EACH ESTABLISHED BY INTRAPERITONEAL INJECTION WITH CIGARETTE SMOKE EXTRACT (CSE) OR INTRATRACHEAL LENTIVIRAL VECTORS INSTILLATION. TUNEL (TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE DUTP NICK END LABELING) ASSAYS WERE USED TO ASSESS APOPTOTIC RATIO IN MICE. THE APOPTOSIS OF HUMAN BRONCHIAL EPITHELIAL CELLS (HBECS) WAS ASSAYED BY FLOW CYTOMETRY. HBO1, B-CELL LYMPHOMA-2 (BCL-2), AND H3K14AC PROTEIN EXPRESSION WERE DETECTED BY WESTERN BLOTTING. HBO1 MRNA EXPRESSION WAS MEASURED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. RESULTS: PROTEIN EXPRESSION OF HBO1 WAS DECREASED SIGNIFICANTLY IN LUNG TISSUE FROM COPD PATIENTS AND CSE-TREATED EMPHYSEMA MOUSE MODELS. OVEREXPRESSION OF HBO1 ATTENUATED CSE-INDUCED EMPHYSEMATOUS CHANGES, AS WELL AS APOPTOSIS IN THE LUNGS OF COPD MICE. IN VITRO, THE HBO1 PROTEIN DEGRADED IN A TIME- AND DOSE-DEPENDENT COURSE WITH CSE TREATMENT. WITH FLOW CYTOMETRY, WE PROVED THAT HBO1 COULD REVERSE THE APOPTOSIS OF HBECS INDUCED BY CSE. FURTHERMORE, HBO1 OVEREXPRESSION PROMOTED THE EXPRESSION OF ANTI-APOPTOTIC BCL-2 PROTEIN AND ENHANCED H3K14 ACETYLATION IN AIRWAY EPITHELIAL CELLS. CONCLUSION: THESE FINDINGS DEMONSTRATE THAT THE KEY HISTONE MODULATOR HBO1 PLAYS A PROTECTIVE ROLE IN COPD PATHOGENESIS THAT MAY SHED LIGHT ON POTENTIAL THERAPEUTIC TARGETS TO INHIBIT THE PROGRESS OF COPD.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
18 1777  28 EDIBLE BLUE-GREEN ALGAE REDUCE THE PRODUCTION OF PRO-INFLAMMATORY CYTOKINES BY INHIBITING NF-KAPPAB PATHWAY IN MACROPHAGES AND SPLENOCYTES. BACKGROUND: CHRONIC INFLAMMATION CONTRIBUTES TO THE DEVELOPMENT OF PATHOLOGICAL DISORDERS INCLUDING INSULIN RESISTANCE AND ATHEROSCLEROSIS. IDENTIFICATION OF ANTI-INFLAMMATORY NATURAL PRODUCTS CAN PREVENT THE INFLAMMATORY DISEASES. METHODS: ANTI-INFLAMMATORY EFFECTS OF BLUE-GREEN ALGAE (BGA), I.E., NOSTOC COMMUNE VAR. SPHAEROIDES KUTZING (NO) AND SPIRULINA PLATENSIS (SP), WERE COMPARED IN RAW 264.7 AND MOUSE BONE MARROW-DERIVED MACROPHAGES (BMM) AS WELL AS SPLENOCYTES FROM APOLIPOPROTEIN E KNOCKOUT (APOE(-/-)) MICE FED BGA. RESULTS: WHEN MACROPHAGES PRETREATED WITH 100MUG/ML NO LIPID EXTRACT (NOE) OR SP LIPID EXTRACT (SPE) WERE ACTIVATED BY LIPOPOLYSACCHARIDE (LPS), EXPRESSION AND SECRETION OF PRO-INFLAMMATORY CYTOKINES, SUCH AS TUMOR NECROSIS FACTOR ALPHA (TNFALPHA), INTERLEUKIN 1BETA (IL-1BETA), AND IL-6, WERE SIGNIFICANTLY REPRESSED. NOE AND SPE ALSO SIGNIFICANTLY REPRESSED THE EXPRESSION OF TNFALPHA AND IL-1BETA IN BMM. LPS-INDUCED SECRETION OF IL-6 WAS LOWER IN SPLENOCYTES FROM APOE(-/-) FED AN ATHEROGENIC DIET CONTAINING 5% NO OR SP FOR 12WEEKS. IN RAW 264.7 MACROPHAGES, NOE AND SPE MARKEDLY DECREASED NUCLEAR TRANSLOCATION OF NF-KAPPAB. THE DEGREE OF REPRESSION OF PRO-INFLAMMATORY GENE EXPRESSION BY ALGAL EXTRACTS WAS MUCH STRONGER THAN THAT OF SN50, AN INHIBITOR OF NF-KAPPAB NUCLEAR TRANSLOCATION. TRICHOSTATIN A, A PAN HISTONE DEACETYLASE INHIBITOR, INCREASED BASAL EXPRESSION OF IL-1BETA AND ATTENUATED THE REPRESSION OF THE GENE EXPRESSION BY SPE. SPE SIGNIFICANTLY DOWN-REGULATED MRNA ABUNDANCE OF 11 HDAC ISOFORMS, CONSEQUENTLY INCREASING ACETYLATED HISTONE 3 LEVELS. CONCLUSION: NOE AND SPE REPRESS PRO-INFLAMMATORY CYTOKINE EXPRESSION AND SECRETION IN MACROPHAGES AND SPLENOCYTES VIA INHIBITION OF NF-KAPPAB PATHWAY. HISTONE ACETYLATION STATE IS LIKELY INVOLVED IN THE INHIBITION. GENERAL SIGNIFICANCE: THIS STUDY UNDERSCORES NATURAL PRODUCTS CAN EXERT ANTI-INFLAMMATORY EFFECTS BY EPIGENETIC MODIFICATIONS SUCH AS HISTONE ACETYLATION.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
19 1210  26 COX2 ENHANCES NEOVASCULARIZATION OF INFLAMMATORY TENOCYTES THROUGH THE HIF-1ALPHA/VEGFA/PDGFB PATHWAY. TENDON INJURIES ARE AMONG THE MOST CHALLENGING IN ORTHOPEDICS. DURING THE EARLY TENDON REPAIR, NEW BLOOD VESSEL FORMATION IS NECESSARY. HOWEVER, EXCESSIVE ANGIOGENESIS ALSO EXACERBATES SCAR FORMATION, LEADING TO PAIN AND DYSFUNCTION. A SIGNIFICANTLY WORSE OUTCOME WAS ASSOCIATED WITH HIGHER EXPRESSION LEVELS OF HYPOXIA-INDUCIBLE FACTOR-1 ALPHA (HIF-1ALPHA), AND ITS TRANSCRIPTIONAL TARGETS VASCULAR ENDOTHELIAL GROWTH FACTOR A (VEGFA) AND PLATELET-DERIVED GROWTH FACTOR B (PDGFB), BUT THE UNDERLYING MOLECULAR MECHANISMS REMAIN UNCLEAR. IN THIS STUDY, LIPOPOLYSACCHARIDE (LPS) WAS USED TO INDUCE AN INFLAMMATORY RESPONSE IN TENOCYTES. LPS INCREASED THE TENOCYTES' INFLAMMATORY FACTOR COX2 EXPRESSION AND ACTIVATED THE HIF-1ALPHA/VEGFA/PDGFB PATHWAY. MOREOVER, THE CONDITIONED MEDIUM FROM THE TENOCYTES BOOSTED RAT AORTIC VASCULAR ENDOTHELIAL CELL (RAOEC) ANGIOGENESIS. FURTHERMORE, TRICHOSTATIN A (TSA), AN INHIBITOR OF HISTONE DEACETYLASE, WAS USED TO TREAT INFLAMMATORY TENOCYTES. THE EXPRESSION LEVELS OF HIF-1ALPHA AND ITS TRANSCRIPTIONAL TARGETS VEGFA AND PDGFB DECREASED, RESULTING IN RAOEC ANGIOGENESIS INHIBITION. FINALLY, THE DUAL-LUCIFERASE REPORTER GENE ASSAY AND CHROMATIN IMMUNOPRECIPITATION (CHIP) ASSAY PROVED THAT THE HIF-1ALPHA/PDGFB PATHWAY PLAYED A MORE CRITICAL ROLE IN TENOCYTE ANGIOGENESIS THAN THE HIF-1ALPHA/VEGFA PATHWAY. TSA COULD ALLEVIATE ANGIOGENESIS MAINLY THROUGH EPIGENETIC REGULATION OF THE HIF-1ALPHA/PDGFB PATHWAY. TAKEN TOGETHER, TSA MIGHT BE A PROMISING ANTI-ANGIOGENESIS DRUG FOR ABNORMAL ANGIOGENESIS, WHICH IS INDUCED BY TENDON INJURIES.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
20 3724  31 INHIBITION OF EZH2 AMELIORATES CARTILAGE ENDPLATE DEGENERATION AND ATTENUATES THE PROGRESSION OF INTERVERTEBRAL DISC DEGENERATION VIA DEMETHYLATION OF SOX-9. BACKGROUND: CARTILAGINOUS ENDPLATE (CEP) DEGENERATION IS CONSIDERED AS ONE OF THE MAJOR CAUSES OF INTERVERTEBRAL DISC DEGENERATION (IVDD) WHICH CAUSES LOW BACK PAIN. RECENT STUDIES HAVE PROVED THAT EPIGENETIC ALTERATION IS INVOLVED IN A VARIETY OF DISEASES. THIS WORK EXPLORED THE ROLE OF HISTONE METHYLTRANSFERASE ENHANCER OF ZESTE HOMOLOGUE 2 (EZH2) IN CEP DEGENERATION, AS WELL AS ITS UNDERLYING EPIGENETIC MECHANISMS, AND CONFIRMED THE EFFECT OF EZH2 KNOCKDOWN ON DELAYING IVDD DEVELOPMENT. METHODS: WESTERN BLOTTING, IMMUNOFLUORESCENCE STAINING, AND CHIP ASSAY WERE APPLIED TO DEMONSTRATE THE MOLECULAR MECHANISM OF EZH2 IN CEP TISSUE. THE THERAPEUTIC POTENTIAL OF EZH2 WAS INVESTIGATED USING PUNCTURE-INDUCED RAT MODELS. FINDINGS: THE EZH2 EXPRESSION WAS UPREGULATED IN HUMAN AND RAT CEP TISSUE. IT WAS ALSO FOUND THAT THE OVEREXPRESSION OF EZH2 SUPPRESSED THE EXPRESSION OF COLLAGEN II, AGGRECAN AND SOX-9, AND PROMOTED THE EXPRESSION OF ADTAMTS5 AND MMP13 IN RAT ENDPLATE CHONDROCYTES (EPCS), WHICH COULD BE REVERSED BY EZH2 SILENCING. THE CORRELATION BETWEEN EZH2 AND SOX-9 WAS FURTHER EXPLORED, WHILE OVEREXPRESSION OF SOX-9 COULD REVERSE THE EFFECT OF EZH2 IN RAT EPCS. MOREOVER, INHIBITION OF EZH2 UPREGULATED THE LEVEL OF SOX-9 BY DEMETHYLATING H3K27ME3 AT SOX-9 PROMOTER SITES, REVEALING THE REGULATORY MECHANISM OF EZH2 ON SOX-9. MEANWHILE, PUNCTURE-INDUCED RAT MODELS SHOWED THAT EZH2 KNOCKDOWN EXERTED A PROTECTIVE EFFECT ON CEP AND DISC DEGENERATION. INTERPRETATION: THIS STUDY REVEALS THAT EZH2 INHIBITION IS A PROMISING STRATEGY FOR MITIGATING THE SYMPTOMS AND PROGRESSION OF IVDD. FUNDING: THIS STUDY WAS FUNDED BY THE NATURAL SCIENCE FOUNDATION OF ZHEJIANG PROVINCE (Y16H060034). AUTHORS DECLARE THAT THE FUNDERS HAD NO INVOLVEMENT IN THE STUDY DESIGN, DATA ANALYSIS AND INTERPRETATION OF THE RESULTS.	2019