1   479 157 ARSENIC TRIOXIDE INHIBITS DNA METHYLTRANSFERASE AND RESTORES TMS1 GENE EXPRESSION IN K562 CELLS. BACKGROUND: GENE SILENCING ASSOCIATED WITH ABERRANT METHYLATION OF PROMOTER REGION CPG ISLANDS IS AN ACQUIRED EPIGENETIC ALTERATION THAT SERVES AS AN ALTERNATIVE TO GENETIC DEFECTS IN THE INACTIVATION OF TUMOR SUPPRESSOR GENES IN HUMAN CANCERS. THE DEMETHYLATING, DOSE-DEPENDENT EFFECT OF ARSENIC TRIOXIDE (AS2O3) ON SEVERAL TUMOR-RELATED GENES HAS ALREADY BEEN POSTULATED. HOWEVER, WHETHER SUCH A DEMETHYLATING EFFECT ALSO APPLIES TO THE TMS1 GENE IN CHRONIC MYELOID LEUKEMIA CELL LINE K562 CELLS HAS NOT BEEN STUDIED SO FAR. THE AIM OF THE PRESENT STUDY WAS TO DETECT THE METHYLATION STATUS OF THE TMS1 GENE IN K562 CELLS AND THE DEMETHYLATION EFFECT OF AS2O3 ON TMS1 AS WELL AS TMS1 APOPTOSIS-ASSOCIATED PROTEIN BCL-2/BAX AND DNA METHYLTRANSFERASE (DNMT) EXPRESSION. METHODS: TMS1 MRNA EXPRESSION IN K562 CELLS AND NORMAL BONE MARROW WAS DETERMINED BY REVERSE TRANSCRIPTION (RT) POLYMERASE CHAIN REACTION (PCR), AND THE DNA METHYLATION STATUS OF THE TMS1 PROMOTER IN K562 CELLS TREATED WITH DIFFERENT CONCENTRATIONS OF AS2O3 FOR 48 H WAS DETERMINED BY METHYLATION-SPECIFIC PCR. RT-PCR AND WESTERN BLOT WERE USED TO DETECT TMS1 AND DNMT EXPRESSION. WE ALSO ASSESSED TMS1-ASSOCIATED APOPTOSIS PROTEIN BCL-2/BAX EXPRESSION BY WESTERN BLOT AND APOPTOSIS RATES BY FLOW CYTOMETRY USING ANNEXIN V/PROPIDIUM IODIDE DOUBLE STAINING. RESULTS: IN K562 CELLS, TMS1 WAS COMPLETELY METHYLATED AND BOTH TMS1 MRNA AND PROTEIN SHOWED A LOW EXPRESSION, BUT 2 MUMOL/L AS2O3 COULD SIGNIFICANTLY RESTORE THE EXPRESSION OF THE TMS1 GENE BOTH AT MRNA AND PROTEIN LEVEL (P < 0.01) BY FULLY REVERSING DNA METHYLATION. AS2O3 DECREASED MRNA AND PROTEIN EXPRESSION OF DNMT1 (P < 0.05) IN A DOSE-DEPENDENT MANNER. FLOW CYTOMETRY SHOWED THAT IN THE EXPERIMENTAL GROUP (2 MUMOL/L AS2O3), CELL APOPTOSIS WAS SIGNIFICANTLY INCREASED COMPARED WITH THE CONTROL GROUP (NO AS2O3; P < 0.05). IN THE EXPERIMENTAL GROUP, WESTERN BLOT SHOWED THAT THE EXPRESSION OF THE ANTI-APOPTOTIC PROTEIN BCL-2 WAS SIGNIFICANTLY DECREASED; HOWEVER, THE PROAPOPTOTIC PROTEIN BAX WAS MARKEDLY INCREASED AND THE BCL-2/BAX RATIO WAS MARKEDLY REDUCED (P < 0.01). CONCLUSIONS: AS2O3 COULD RESTORE THE EXPRESSION OF TMS1 BY INHIBITING DNMT TO REVERSE THE HYPERMETHYLATION AND INDUCED APOPTOSIS OF K562 CELLS BY DOWNREGULATION OF BCL-2/BAX EXPRESSION.	2015	
                                                                                                                                                                                                 
2  6455  52 THYMOQUINONE ENHANCES APOPTOSIS OF K562 CHRONIC MYELOID LEUKEMIA CELLS THROUGH HYPOMETHYLATION OF SHP-1 AND INHIBITION OF JAK/STAT SIGNALING PATHWAY. THE EPIGENETIC SILENCING OF TUMOR SUPPRESSOR GENES (TSGS) IS CRITICAL IN THE DEVELOPMENT OF CHRONIC MYELOID LEUKEMIA (CML). SHP-1 FUNCTIONS AS A TSG AND NEGATIVELY REGULATES JAK/STAT SIGNALING. ENHANCEMENT OF SHP-1 EXPRESSION BY DEMETHYLATION PROVIDES MOLECULAR TARGETS FOR THE TREATMENT OF VARIOUS CANCERS. THYMOQUINONE (TQ), A CONSTITUENT OF NIGELLA SATIVA SEEDS, HAS SHOWN ANTI-CANCER ACTIVITIES IN VARIOUS CANCERS. HOWEVER, TQS EFFECT ON METHYLATION IS NOT FULLY CLEAR. THEREFORE, THE AIM OF THIS STUDY IS TO ASSESS TQS ABILITY TO ENHANCE THE EXPRESSION OF SHP-1 THROUGH MODIFYING DNA METHYLATION IN K562 CML CELLS. THE ACTIVITIES OF TQ ON CELL CYCLE PROGRESSION AND APOPTOSIS WERE EVALUATED USING A FLUOROMETRIC-RED CELL CYCLE ASSAY AND ANNEXIN V-FITC/PI, RESPECTIVELY. THE METHYLATION STATUS OF SHP-1 WAS STUDIED BY PYROSEQUENCING ANALYSIS. THE EXPRESSION OF SHP-1, TET2, WT1, DNMT1, DNMT3A, AND DNMT3B WAS DETERMINED USING RT-QPCR. THE PROTEIN PHOSPHORYLATION OF STAT3, STAT5, AND JAK2 WAS ASSESSED USING JESS WESTERN ANALYSIS. TQ SIGNIFICANTLY DOWNREGULATED THE DNMT1 GENE, DNMT3A GENE, AND DNMT3B GENE AND UPREGULATED THE WT1 GENE AND TET2 GENE. THIS LED TO HYPOMETHYLATION AND RESTORATION OF SHP-1 EXPRESSION, RESULTING IN INHIBITION OF JAK/STAT SIGNALING, INDUCTION OF APOPTOSIS, AND CELL CYCLE ARREST. THE OBSERVED FINDINGS IMPLY THAT TQ PROMOTES APOPTOSIS AND CELL CYCLE ARREST IN CML CELLS BY INHIBITING JAK/STAT SIGNALING VIA RESTORATION OF THE EXPRESSION OF JAK/STAT-NEGATIVE REGULATOR GENES.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
3  2326  55 EPIGENETIC REGULATION OF HOTAIR IN ADVANCED CHRONIC MYELOID LEUKEMIA. PURPOSE: CHRONIC MYELOID LEUKEMIA (CML) ACCOUNTS FOR ~10% OF LEUKEMIA CASES, AND ITS PROGRESSION INVOLVES EPIGENETIC GENE REGULATION. THIS STUDY INVESTIGATED EPIGENETIC REGULATION OF HOTAIR AND ITS TARGET MICRORNA, MIR-143, IN ADVANCED CML. PATIENTS AND METHODS: WE FIRST ISOLATED BONE MARROW MONONUCLEAR CELLS FROM 70 PATIENTS WITH DIFFERENT PHASES OF CML AND FROM HEALTHY DONORS AS NORMAL CONTROL; WE ALSO CULTURED K562 AND KCL22 CELLS, TREATED WITH DEMETHYLATION DRUG; MTT ASSAY, FLOW CYTOMETRY, QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QPCR), METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP), WESTERN BLOT, LUCIFERASE ASSAY, RNA PULL-DOWN ASSAY AND RNA-BINDING PROTEIN IMMUNOPRECIPITATION (RIP) ASSAY WERE PERFORMED. RESULT: AS MEASURED BY QPCR, HOTAIR EXPRESSION IN K562 CELLS, KCL22 CELLS, AND SAMPLES FROM CASES OF ADVANCED-STAGE CML INCREASED WITH LEVELS OF SEVERAL DNA METHYLTRANSFERASES AND HISTONE DEACETYLATES, INCLUDING DNMT1, DNMT3A, HDAC1, EZH2, AND LSD1, AND MIR-143 LEVELS WERE DECREASED AND HOTAIR LEVELS WERE INCREASED. TREATMENT WITH 5-AZACYTIDINE, A DNA METHYLATION INHIBITOR, DECREASED DNMT1, DNMT3A, HDAC1, EZH2, LSD1 MRNA, PROTEIN LEVELS, AND HOTAIR MRNA LEVELS BUT INCREASED MIR-143 LEVELS. HOTAIR KNOCKDOWN AND MIR-143 OVEREXPRESSION BOTH INHIBITED PROLIFERATION AND PROMOTED APOPTOSIS IN KCL22 AND K562 CELLS THROUGH THE PI3K/AKT PATHWAY. RNA PULL-DOWN, MASS SPECTROMETRY, AND RIP ASSAYS SHOWED THAT HOTAIR INTERACTED WITH EZH2 AND LSD1. A DUAL-LUCIFERASE ASSAY DEMONSTRATED THAT HOTAIR INTERACTED WITH MIR-143. CONCLUSION: OUR FINDINGS DEMONSTRATE THE KEY EPIGENETIC INTERACTIONS OF HOTAIR RELATED TO CML PROGRESSION AND SUGGEST HOTAIR AS A POTENTIAL THERAPEUTIC TARGET FOR ADVANCED CML. FURTHERMORE, OUR RESULTS SUPPORT THE USE OF DEMETHYLATION DRUGS AS A CML TREATMENT STRATEGY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
4  1272  51 CYTOTOXIC ACTIVITY OF VALPROIC ACID ON PRIMARY CHRONIC LYMPHOCYTIC LEUKEMIA CELLS. BACKGROUND: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS THE MOST COMMON ADULT LEUKEMIA IN WESTERN CIVILIZATION. THE ACCUMULATION OF CD5+CD19+ B LYMPHOCYTES IN PERIPHERAL BLOOD IS DUE TO A DEFECT IN THE APOPTOTIC PATHWAY RATHER THAN EXCESSIVE PROLIFERATION IN THE BONE MARROW AND LYMPH NODES. DESPITE A NUMBER OF TREATMENTS, CLL REMAINS AN INCURABLE DISEASE. VALPROIC ACID (VPA) ACTIVITY, AS A HISTONE DEACETYLASE INHIBITOR, COULD RESTORE THE EPIGENETIC CHANGES UNDERLYING THE PATHOGENESIS OF CLL AND THUS INDUCE CELL DEATH. OBJECTIVES: IN THE PRESENT STUDY WE HYPOTHESIZED THAT VPA COULD INDUCE CLL PRIMARY CELLS DEATH THROUGH ACTIVATION OF APOPTOSIS. MATERIAL AND METHODS: PERIPHERAL BLOOD SAMPLES WERE OBTAINED FROM 53 CLL PATIENTS. PERIPHERAL BLOOD MONONUCLEAR CELLS WERE ISOLATED THROUGH DENSITY GRADIENT CENTRIFUGATION AND WERE THE SUBJECT OF A 24-HOUR CELL CULTURE WITH 10 MM OF VPA. THE CYTOTOXIC EFFECT OF VPA WAS EVALUATED WITH AN XTT TEST AND THEREAFTER CONFIRMED USING ANNEXIN V-FITC/PI STAINING AND FLOW CYTOMETRY TECHNIQUES. RESULTS: IN THIS STUDY, A MEDIAN VPA CYTOTOXICITY OF 13.88% WITH A RANGE OF 0-54.65% WAS OBSERVED. ANNEXIN V/PI STAINING CONFIRMED THAT THE DEMONSTRATED CYTOTOXICITY WAS CAUSED BY INCREASED APOPTOSIS IN THE VPA TREATED CELLS AS COMPARED TO CONTROL CELLS. STATISTICAL ANALYSIS SHOWED THAT VPA'S EFFECT ON CLL CELLS DEPENDS ON LACTATE DEHYDROGENASE SERUM LEVELS, BUT IS INDEPENDENT OF ALL OTHER PROGNOSTIC MARKERS. CONCLUSIONS: THE RESULTS OF THE PRESENT EXPERIMENTS FOUND THAT VPA AT A CLINICALLY APPLICABLE CONCENTRATION SIGNIFICANTLY INDUCES APOPTOSIS INDEPENDENTLY OF THE DISEASE STAGE AND MIGHT BE A VALUABLE THERAPEUTIC AGENT FOR ALL CLL PATIENTS.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
5  5459  46 RESEARCH ON THE EPIGENETIC REGULATION MECHANISM OF THE PTPN6 GENE IN ADVANCED CHRONIC MYELOID LEUKAEMIA. PTPN6, A TYROSINE PHOSPHATASE PROTEIN, PLAYS A NEGATIVE ROLE IN CELL SIGNAL TRANSDUCTION AND IS NEGATIVELY CORRELATED WITH TUMOUR FORMATION AND GROWTH. HOWEVER, EPIGENETIC REGULATION MECHANISM OF THE PTPN6 GENE IN ADVANCED CHRONIC MYELOID LEUKAEMIA (CML) REMAINS UNCLEAR. THIS STUDY INVESTIGATED BONE MARROW OR BLOOD SAMPLES FROM 44 CML PATIENTS AND 10 HEALTHY VOLUNTEERS. KCL22 AND K562 CELLS WERE CULTURED AND TREATED WITH DEMETHYLATION DRUGS AND HISTONE DEACETYLASE INHIBITORS. REAL TIME QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR), METHYLATION-SPECIFIC PCR, BISULFITE SEQUENCING PCR, WESTERN BLOTTING, CO-IMMUNOPRECIPITATION AND CHROMATIN IMMUNOPRECIPITATION (CHIP) WAS PERFORMED. PTPN6 WAS DOWN-REGULATED IN CELL LINES AND PATIENTS WITH ADVANCED PHASE CML, WHEREAS DNMT1, DNMT3A, MECP2, MBD2 AND HDAC1 WERE UP-REGULATED. TREATMENT WITH 5-AZACYTIDINE, DECITABINE, SODIUM VALPROATE AND LBH589 INCREASED PTPN6 EXPRESSION, BUT DECREASED THAT OF DNMT1, DNMT3A, MECP2, MBD2 AND HDAC1. IMMUNOPRECIPITATION AND MASS SPECTROMETRY SHOWED THAT HDAC1 COMBINED DIRECTLY WITH PTPN6. CHIP-SEQ SHOWED THAT HDAC1 DID NOT COMBINE WITH THE PROMOTER REGION OF PTPN6, WHILE MAPK, AKT, STAT5, JAK2 AND MYC PROMOTER REGIONS ALL COMBINED WITH HDAC1. PTPN6 IS ASSOCIATED WITH PROGRESSION OF CML. LOW EXPRESSION LEVEL OF PTPN6 WAS ASSOCIATED WITH DNA METHYLATION AND REGULATED BY HISTONE ACETYLATION. HDAC1 PARTICIPATES IN THE REGULATION OF PTPN6.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
6  4601  59 NDRG2 MRNA LEVELS AND MIR-28-5P AND MIR-650 ACTIVITY IN CHRONIC LYMPHOCYTIC LEUKEMIA. BACKGROUND: NDRG2 IS IDENTIFIED AS A TUMOR SUPPRESSOR GENE IN MANY TUMORS, AND FUNCTIONS IN CELL PROLIFERATION, DIFFERENTIATION AND APOPTOSIS. RECENT DATA INDICATE THAT NDRG2 EXPRESSION IS UP-REGULATED BY TP53. MOREOVER, PROPOSED MECHANISMS OF NDRG2 INACTIVATION INCLUDE EPIGENETIC SILENCING OF THE NDRG2 PROMOTER AND DOWN-REGULATION BY MICRORNAS (MIRNAS). HOWEVER, FEW STUDIES HAVE EVER BEEN DONE ON THE ROLE OF NDRG2 AND THE NDRG2-REGULATING MIRNAS INTERFERENCE IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). METHODS: NDRG2 AND MICRORNAS MRNA LEVELS IN CLL SUBJECTS WERE ASSESSED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QRT-PCR). THE DUAL-LUCIFERASE REPORTER ASSAY WAS PERFORMED TO DETERMINE NDRG2-RELATED MIRNAS. LOW EXPRESSION OF MATURE EXOGENOUS MIRNAS IN CLL CELLS WAS ESTABLISHED BY TRANSIENT TRANSFECTION. NDRG2 PROTEIN LEVELS IN CLL CELLS WERE DETECTED BY WESTERN BLOT. IN ADDITION, FLOW CYTOMETRY WAS CONDUCTED TO EXAMINE THE APOPTOSIS OF CLL CELLS. RESULTS: LOWER EXPRESSION OF NDRG2 WAS FOUND IN THE B-CELLS FROM 102 CLL PATIENTS COMPARED THE 40 NORMAL SUBJECTS (P < 0.001). PATIENTS WITH ADVANCED BINET STAGE (P = 0.001), HIGH LACTATE DEHYDROGENASE (LDH) LEVEL (P = 0.036), UN-MUTATED IMMUNOGLOBULIN HEAVY CHAIN VARIABLE REGION GENE (IGHV) (P = 0.004) AND THOSE WITH P53 ABERRATIONS (P < 0.001) HAD A MARKEDLY LOWER LEVELS OF NDRG2 MRNA. THIS DECREASE WAS ASSOCIATED WITH BRIEFER TIME-TO-TREATMENT (P = 0.001) AND POORER SURVIVAL (P < 0.001). HIGH EXPRESSION OF MIR-28-5P AND MIR-650 WAS ASSOCIATED WITH BINET B/C STAGE (P = 0.044) AND IGHV UN-MUTATED (P = 0.011), AS WELL AS BINET B/C STAGE (P = 0.013) AND P53 ABERRATIONS (P = 0.037), RESPECTIVELY. INHIBITION OF MIR-28-5P OR MIR-650 COULD INDUCE MORE APOPTOSIS IN CLL CELLS WITH GERMLINE TP53. CONCLUSIONS: NDRG2 MRNA LEVELS MIGHT BE A USEFUL PROGNOSTIC VARIABLE FOR PATIENTS OF CLL AND UP-REGULATING NDRG2 TRANSCRIPTION MAY BE A THERAPY APPROACH IN CLL WITHOUT P53 ABERRATIONS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
7  1905  47 ENHANCER OF ZESTE HOMOLOG 2 CONTRIBUTES TO APOPTOSIS BY INACTIVATING JANUS KINASE 2/ SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION SIGNALING IN INFLAMMATORY BOWEL DISEASE. BACKGROUND: INFLAMMATORY BOWEL DISEASE (IBD) IS A PREVALENT WORLDWIDE HEALTH PROBLEM FEATURED BY RELAPSING, CHRONIC GASTROINTESTINAL INFLAMMATION. ENHANCER OF ZESTE HOMOLOG 2 (EZH2) IS A CRITICAL EPIGENETIC REGULATOR IN DIFFERENT PATHOLOGICAL MODELS, SUCH AS CANCER AND INFLAMMATION. HOWEVER, THE ROLE OF EZH2 IN THE IBD DEVELOPMENT IS STILL OBSCURE. AIM: TO EXPLORE THE EFFECT OF EZH2 ON IBD PROGRESSION AND THE UNDERLYING MECHANISM. METHODS: THE IBD MOUSE MODEL WAS CONDUCTED BY ADDING DEXTRAN SODIUM SULFATE (DSS), AND THE EFFECT OF EZH2 ON DSS-INDUCED COLITIS WAS ASSESSED IN THE MODEL. THE FUNCTION OF EZH2 IN REGULATING APOPTOSIS AND PERMEABILITY WAS EVALUATED BY ANNEXIN V-FITC APOPTOSIS DETECTION KIT, TRANSEPITHELIAL ELECTRICAL RESISTANCE ANALYSIS, AND WESTERN BLOT ANALYSIS OF RELATED MARKERS, INCLUDING ZONA OCCLUDENS 1, CLAUDIN-5, AND OCCLUDIN, IN NCM460 AND FETAL HUMAN COLON (FHC) CELLS. THE MECHANICAL INVESTIGATION WAS PERFORMED BY QUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION, WESTERN BLOT ANALYSIS, AND CHROMATIN IMMUNOPRECIPITATION ASSAYS. RESULTS: THE COLON LENGTH WAS INHIBITED IN THE DSS-TREATED MICE AND WAS ENHANCED BY THE EZH2 DEPLETION IN THE SYSTEM. DSS TREATMENT CAUSED A DECREASED HISTOLOGICAL SCORE IN THE MICE, WHICH WAS REVERSED BY EZH2 DEPLETION. THE INFLAMMATORY CYTOKINES, SUCH AS TUMOR NECROSIS FACTOR-ALPHA, INTERLEUKIN-6, AND INTERLEUKIN-1BETA, WERE INDUCED IN THE DSS-TREATED MICE, IN WHICH THE DEPLETION OF EZH2 COULD REVERSE THIS EFFECT. MOREOVER, THE TUMOR NECROSIS FACTOR-ALPHA TREATMENT INDUCED THE APOPTOSIS OF NCM460 AND FHC CELLS, IN WHICH EZH2 DEPLETION COULD REVERSE THIS EFFECT IN THE CELLS. MOREOVER, THE DEPLETION OF EZH2 ATTENUATED PERMEABILITY OF COLONIC EPITHELIAL CELLS. MECHANICALLY, THE DEPLETION OF EZH2 OR EZH2 INHIBITOR GSK343 WAS ABLE TO ENHANCE THE EXPRESSION AND THE PHOSPHORYLATION OF JANUS KINASE 2 (JK2) AND SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION IN THE NCM460 AND FHC CELLS. SPECIFICALLY, EZH2 INACTIVATED JAK2 EXPRESSION BY REGULATING HISTONE H3K27ME3. JAK2 INHIBITOR TG101348 WAS ABLE TO REVERSE EZH2 KNOCKDOWN-MEDIATED COLONIC EPITHELIAL CELL PERMEABILITY AND APOPTOSIS. CONCLUSION: THUS, WE CONCLUDED THAT EZH2 CONTRIBUTED TO APOPTOSIS AND INFLAMMATORY RESPONSE BY INACTIVATING JAK2/ SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION SIGNALING IN IBD. EZH2 MAY BE APPLIED AS A POTENTIAL TARGET FOR IBD THERAPY.	2021	

8  1613  47 DNA METHYLTRANSFERASE 1 MEDIATED ABERRANT METHYLATION AND SILENCING OF SHP-1 GENE IN CHRONIC MYELOGENOUS LEUKEMIA CELLS. INTRODUCTION: EXTENSIVE STUDIES ON SHP-1 PROTEIN AND SHP-1 MRNA REVEALED THAT THE DIMINISHMENT OR ABOLISHMENT OF THE EXPRESSION OF SHP-1 IN LEUKEMIAS/LYMPHOMAS WAS DUE TO ABERRANT PROMOTER METHYLATION. THUS FAR, THE MECHANISM OF EPIGENETIC SILENCING OF THE SHP-1 TYROSINE PHOSPHATASE GENE THAT OCCURS IN CHRONIC MYELOGENOUS LEUKEMIA CELLS REMAINS POORLY UNDERSTOOD. METHODS: THE EXPRESSIONS OF THE TARGET MOLECULES WERE DETERMINED BY QUANTITATIVE REAL TIME PCR AND WESTERN BLOT, RESPECTIVELY. BISULFITE SEQUENCING PCR WAS USED TO DETECT METHYLATION STATUS OF DNA CPG. THE LENTIVIRAL VECTORS WERE APPLIED TO MODIFY GENE EXPRESSION. RESULTS: IN THE PRESENT STUDY, WE FOUND THAT THE PROMOTER 2 OF SHP-1 GENE IS LOCATED BETWEEN POSITIONS FROM -577BP TO +300BP, AND 22 CPG SITES CONTAINED IN POSITIONS -353BP APPROXIMATELY +182BP ARE ABERRANTLY METHYLATED IN K562 CELLS. IN VITRO, WE DEMONSTRATED THAT DNMT1 SILENCING INDUCED DEMETHYLATION OF THE 22 CPG SITES LOCATED IN THE SHP-1 PROMOTER AND RE-EXPRESSION OF SHP-1 GENE IN K562 CELLS. MOREOVER, WE PROVED THAT THE EXPRESSION LEVELS OF DNMT1 AND SHP-1 MRNA AND PROTEIN WERE NEGATIVELY CORRELATED IN K562 CELLS AND BM ASPIRATES MONONUCLEAR CELLS FROM CML PATIENTS. CONCLUSION: COLLECTIVELY, THESE RESULTS INDICATE THAT DNMT1 MEDIATES ABERRANT METHYLATION AND SILENCING OF SHP-1 GENE IN CHRONIC MYELOGENOUS LEUKEMIA CELLS, AND PROVIDE A NOVEL THERAPEUTIC TARGET FOR CML.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
9   574  42 BCR/ABL INCREASES EZH2 LEVELS WHICH REGULATES XIAP EXPRESSION VIA MIRNA-219 IN CHRONIC MYELOID LEUKEMIA CELLS. IN THIS STUDY, WE SHOWED THAT THE LEVELS OF EZH2 IN BONE MARROW MONONUCLEAR CELLS (BMMNCS) ISOLATED FROM INDIVIDUALS WITH CHRONIC MYELOID LEUKEMIA (CML) (N=12) WERE SIGNIFICANTLY GREATER THAN THOSE IN BMMNCS ISOLATED FROM HEALTHY VOLUNTEERS (N=6) AS WELL AS INDIVIDUALS WITH PHILADELPHIA CHROMOSOME-NEGATIVE MYELOPROLIFERATIVE NEOPLASMS. LENTIVIRAL TRANSDUCTION OF THE BCR/ABL GENE IN BA/F3 CELLS INCREASED EZH2 LEVELS IN PARALLEL WITH PHOSPHORYLATION OF STAT5. NOTABLY, CHROMATIN IMMUNOPRECIPITATION ASSAYS SHOWED THAT STAT5A BOUND TO A PROMOTER REGION OF THE EZH2 GENE, RESULTING IN AN INCREASE IN THE TRANSCRIPTIONAL ACTIVITY OF EZH2 IN LEUKEMIA CELLS. IMPORTANTLY, DOWNREGULATION OF EZH2 BY SHORT HAIRPIN RNAS (SHRNAS) INHIBITED THE EXPRESSION OF XIAP AND INCREASED THE MIR-219 LEVELS ASSOCIATED WITH A DECREASE IN HYPERMETHYLATION OF MIR-219-1 CPG ISLANDS. MOREOVER, OVEREXPRESSION OF MIR-219 DECREASED THE LEVELS OF XIAP IN CML CELLS. SINCE THE 3'-UNTRANSLATED REGION (3'-UTR) OF XIAP CONTAINS MIR219-5P-COMPLEMENTARY BINDING SITE, MIR-219 MIGHT MODULATE THE EXPRESSION OF XIAP THROUGH BINDING OF MIR-219 ON THE 3'-UTR OF XIAP. TAKEN TOGETHER, BCR/ABL POSITIVELY REGULATES THE EXPRESSION OF EZH2 VIA STAT5 SIGNALING. EZH2 MODULATES EPIGENETIC CHANGES AT DNA METHYLATED REGIONS ENCODING MIR-219 AND DOWNREGULATES THE LEVEL OF MIR-219, RESULTING IN UPREGULATION OF XIAP.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
10  5481  39 RESVERATROL-INDUCED APOPTOSIS DEPENDS ON THE LIPID KINASE ACTIVITY OF VPS34 AND ON THE FORMATION OF AUTOPHAGOLYSOSOMES. IN HUMAN COLORECTAL DLD1 CANCER CELLS, THE DIETARY BIOFLAVONOID RESVERATROL (RV) RAPIDLY INDUCED AUTOPHAGY. THIS EFFECT WAS REVERSIBLE (ON REMOVAL OF THE DRUG) AND WAS ASSOCIATED WITH INCREASED EXPRESSION AND CYTOSOLIC REDISTRIBUTION OF THE PROTEINS BECLIN1 AND LC3 II. SUPPLEMENTING THE CELLS WITH ASPARAGINE (ASN) ABROGATED THE BECLIN-DEPENDENT AUTOPHAGY. WHEN APPLIED ACUTELY (2 H), RV WAS NOT TOXIC; HOWEVER, REITERATE CHRONIC (48 H) EXPOSURE TO RV EVENTUALLY LED TO ANNEXIN V- AND TERMINAL DEOXINUCLEOTIDYL TRANSFERASE-MEDIATED DUTP-BIOTIN NICK END LABELING-POSITIVE CELL DEATH. THIS TOXIC EFFECT WAS AUTOPHAGY DEPENDENT, AS IT WAS PREVENTED EITHER BY ASN, BY EXPRESSING A DOMINANT-NEGATIVE LIPID KINASE-DEFICIENT CLASS III PHOSPHOINOSITIDE 3-PHOSPHATE KINASE, OR BY RNA INTERFERENCE KNOCKDOWN OF BECLIN1. LAMP2B SILENCING ABOLISHED THE FUSION OF AUTOPHAGOSOMES WITH LYSOSOMES AND PRESERVED CELL VIABILITY DESPITE THE ONGOING FORMATION OF AUTOPHAGOSOMES IN CELLS CHRONICALLY EXPOSED TO RV. THE PAN-CASPASE INHIBITOR BENZYLOXYCARBONYL-VAL-ALA-ASP-FLUOROMETHYLKETONE INHIBITED RV-INDUCED CELL DEATH, BUT NOT AUTOPHAGY. THESE RESULTS UNCOVER A NOVEL PATHWAY OF RV CYTOTOXICITY IN WHICH AUTOPHAGY PLAYS A DUAL ROLE: (I) AT FIRST, IT ACTS AS A PROSURVIVAL STRESS RESPONSE AND (II) AT A LATER TIME, IT SWITCHES TO A CASPASE-DEPENDENT APOPTOSIS PATHWAY. THE PRESENT DATA ALSO INDICATE THAT GENETIC OR EPIGENETIC INACTIVATION OF AUTOPHAGY PROTEINS IN CANCER CELLS MAY CONFER RESISTANCE TO RV-MEDIATED KILLING.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
11  2440  43 EPIGENETIC SILENCING OF TUMOR SUPPRESSOR LONG NON-CODING RNA BM742401 IN CHRONIC LYMPHOCYTIC LEUKEMIA. BM742401 IS A TUMOR SUPPRESSOR LNCRNA DOWNREGULATED IN GASTRIC CANCER. AS THE PROMOTER REGION AND THE ENTIRE TRANSCRIPT ARE EMBEDDED IN A CPG ISLAND, WE POSTULATED THAT BM742401 IS A TUMOR SUPPRESSOR LNCRNA INACTIVATED BY DNA METHYLATION IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). THE PROMOTER OF BM742401 WAS UNMETHYLATED IN NORMAL CONTROLS INCLUDING THREE EACH OF NORMAL BONE MARROW, PERIPHERAL BLOOD BUFFY COATS, AND CD19-SORTED PERIPHERAL B-CELLS, BUT METHYLATED IN FOUR (57.1%) CLL CELL LINES. METHYLATION OF BM742401 CORRELATED INVERSELY WITH EXPRESSION. IN THE COMPLETELY METHYLATED WAC3CD5+ CLL CELLS, 5-AZA-2'-DEOXYCYTIDINE TREATMENT LED TO PROMOTER DEMETHYLATION AND RE-EXPRESSION OF BM742401 TRANSCRIPT. FUNCTIONALLY, STABLE OVEREXPRESSION OF BM742401 RESULTED IN INHIBITION OF CELLULAR PROLIFERATION AND ENHANCED APOPTOSIS THROUGH CASPASE-9-DEPENDENT INTRINSIC BUT NOT CASPASE-8-DEPENDENT EXTRINSIC APOPTOSIS PATHWAY, SUGGESTING A TUMOR SUPPRESSOR ROLE OF BM742401 IN CLL. IN PRIMARY CLL SAMPLES, METHYLATION OF BM742401 WAS DETECTED IN 43/98 (43.9%) OF PATIENTS. MOREOVER, AMONG CLL PATIENTS WITH STANDARD-RISK CYTOGENETIC ABERRATIONS, METHYLATION OF BM742401 CORRELATED WITH ADVANCED RAI STAGE (>/= STAGE 2)(P = 0.002). FURTHERMORE, BM742401 METHYLATION WAS ASSOCIATED WITH MIR-129-2 METHYLATION (P = 0.05). BM742401 IS A TUMOR SUPPRESSOR LNCRNA FREQUENTLY METHYLATED IN CLL. THE MECHANISM OF BM742401 AS A TUMOR SUPPRESSOR WARRANTS FURTHER STUDIES.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
12  5458  44 RESEARCH ON EPIGENETIC MECHANISM OF SFRP2 IN ADVANCED CHRONIC MYELOID LEUKEMIA. SECRETED FRIZZLED-RELATED PROTEIN 2 (SFRP2) HAS BEEN REPORTED TO ACT AS A TUMOR SUPPRESSORS. THIS STUDY AIMS TO DETECT THE BIOLOGICAL ROLE OF SFRP2 IN ADVANCED CHRONIC MYELOID LEUKEMIA (CML). IN THIS STUDY WE EXAMINED BONE MARROW SAMPLES FROM 45 CML PATIENTS AND 10 HEALTHY DONORS. K562 AND KCL22 CELLS WERE CULTURED AND TREATED WITH DEMETHYLATION DRUG AND HISTONE DEACETYLASE INHIBITOR (HDACI). KCL22 AND K562 CELLS WERE TRANSFECTED WITH LENTIVIRAL VECTOR (LV)-SFRP2, LV-CONTROL. THE CELLS WERE THEN SUBJECTED TO PROLIFERATION AND APOPTOSIS ASSAYS, REAL TIME POLYMERASE CHAIN REACTION (PCR), METHYLATION-SPECIFIC PCR (MSP), WESTERN BLOTTING, CO-IMMUNOPRECIPITATION (COIP) AND CHROMATIN IMMUNOPRECIPITATION (CHIP), WE FOUND THAT SFRP2 WAS DOWN-REGULATED IN THE ACCELERATED AND BLAST PHASE OF CML, WHEREAS, THE LEVELS OF WNT1, WNT3 AND WNT5A WERE UP-REGULATED IN THE ACCELERATED AND BLAST PHASE OF CML. OVEREXPRESSION SFRP2 INHIBITED PROLIFERATION, PROMOTED APOPTOSIS AND ACTIVATED THE WNT PATHWAY. COIP-MS RESULTS SHOWED THAT SFRP2 INTERACTED WITH WNT1 AND WNT5A. CHIP-SEQ RESULT INDICATED THAT THE PROMOTER OF H3K4ME3 AND H3K27ME3 WERE ABLE TO INTERACT WITH SFRP2. IN CONCLUSION, OUR FINDINGS DEMONSTRATED THE SFRP2 ACT AS A POTENTIAL THERAPEUTIC TARGET FOR ADVANCED CML. FURTHERMORE, OUR RESULTS SUPPORT THE USE OF DEMETHYLATION DRUGS AND HDACI AS A POTENTIAL CML TREATMENT STRATEGY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
13  2426  47 EPIGENETIC SILENCING OF LONG NON-CODING RNA BM742401 IN MULTIPLE MYELOMA: IMPACT ON PROGNOSIS AND MYELOMA DISSEMINATION. BACKGROUND: LONG NON-CODING RNA (LNCRNA) BM742401 IS A TUMOR SUPPRESSOR IN GASTRIC CANCER AND CHRONIC LYMPHOCYTIC LEUKEMIA. AS THE PROMOTER AND CODING REGION OF BM742401 ARE FULLY EMBEDDED IN A CPG ISLAND, WE HYPOTHESIZED THAT BM742401 IS A TUMOR SUPPRESSOR LNCRNA EPIGENETICALLY SILENCED BY PROMOTER DNA METHYLATION IN MULTIPLE MYELOMA. METHODS: METHYLATION-SPECIFIC PCR AND QUANTITATIVE BISULFITE PYROSEQUENCING WERE PERFORMED TO DETECT THE METHYLATION OF BM742401 IN NORMAL PLASMA CELLS, MYELOMA CELL LINES AND PRIMARY MYELOMA SAMPLES. THE EXPRESSION OF BM742401 WAS MEASURED BY QRT-PCR. THE FUNCTION OF BM742401 IN MULTIPLE MYELOMA CELLS WAS ANALYZED BY LENTIVIRUS TRANSDUCTION FOLLOWED BY MIGRATION ASSAY. RESULTS: BM742401 METHYLATION WAS DETECTED IN 10 (66.7%) MYELOMA CELL LINES BUT NOT NORMAL PLASMA CELLS, AND INVERSELY CORRELATED WITH EXPRESSION OF BM742401. IN PRIMARY SAMPLES, BM742401 METHYLATION WAS DETECTED IN 3 (12.5%) MONOCLONAL GAMMOPATHY OF UNDETERMINED SIGNIFICANCE, 9 (15.8%) MYELOMA AT DIAGNOSIS AND 8 (17.0%) MYELOMA AT RELAPSE/PROGRESSION. MOREOVER, BM742401 METHYLATION AT DIAGNOSIS WAS ASSOCIATED WITH INFERIOR OVERALL SURVIVAL (MEDIAN OS: 25 VS. 39 MONTHS; P = 0.0496). IN MYELOMA CELL LINE JJN-3, STABLE OVEREXPRESSION OF BM742401 BY LENTIVIRUS TRANSDUCTION RESULTED IN REDUCED CELL MIGRATION (P = 0.0001) BUT NOT IMPACTING CELL DEATH OR PROLIFERATION. CONCLUSIONS: THIS IS THE FIRST REPORT OF TUMOR-SPECIFIC METHYLATION-MEDIATED SILENCING OF BM742401 IN MYELOMA, WHICH IS LIKELY AN EARLY EVENT IN MYELOMAGENESIS WITH ADVERSE IMPACT ON OVERALL SURVIVAL. MOREOVER, BM742401 IS A TUMOR SUPPRESSOR LNCRNA BY INHIBITING MYELOMA CELL MIGRATION, HENCE IMPLICATED IN MYELOMA PLASMA CELL HOMING, METASTASIS AND DISEASE PROGRESSION.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
14  6402  36 THE ROLES OF EPIGENETIC MODIFICATIONS OF PROAPOPTOTIC BID AND BIM GENES IN IMATINIB-RESISTANT CHRONIC MYELOID LEUKEMIA CELLS. IN CHRONIC MYELOID LEUKEMIA (CML), EPIGENETIC MODIFICATIONS SUCH AS PROMOTER HYPERMETHYLATION AND INACTIVE HISTONE MODIFICATION ARE KNOWN MECHANISMS OF DRUG RESISTANCE. IN OUR STUDY, WE INVESTIGATED THE ROLES OF PROMOTER HYPERMETHYLATION OF BIM AND BID GENES AND H3K27ME3 HISTONE MODIFICATION ON IMATINIB RESISTANCE. WE DETECTED HIGHER EXPRESSION LEVELS OF BIM AND BID GENES AND LOWER EXPRESSION LEVELS OF EZH2, EED2, SIRT1, AND SUZ12 GENES IN IMATINIB-RESISTANT K562/IMA-3 CELLS COMPARED TO IMATINIB-NON-RESISTANT K562 CELLS. WHILE WE DETERMINED THE EZH2 AND DNMT ENZYMES AS BOUNDED TO THE PROMOTER OF THE BIM GENE, WE DID NOT DETECT HYPERMETHYLATION OF THIS PROMOTER. WE ALSO FOUND THE H3K27ME3 HISTONE MODIFICATION PROMOTER OF BIM AND BID GENES IN BOTH CELL LINES. IN CONCLUSION, OUR RESULTS SUPPORT THE NOTION THAT DNA PROMOTER METHYLATION MAY BE FORMED INDEPENDENTLY FROM EZH2-H3K27ME3 AND PRO-APOPTOTIC BIM AND BID GENES ARE NOT METHYLLATED IN THE IMATINIB RESISTANCE OF CML CELLS.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
15  6238  47 THE MALIGNANCY SUPPRESSION ROLE OF MIR-23A BY TARGETING THE BCR/ABL ONCOGENE IN CHROMIC MYELOID LEUKEMIA. THE AIM OF THIS STUDY WAS TO INVESTIGATE THE ROLE AND MECHANISM OF MIR-23A IN THE REGULATION OF BCR/ABL AND TO PROVIDE A NEW PROGNOSTIC BIOMARKER FOR CHRONIC MYELOID LEUKEMIA (CML). THE EXPRESSION LEVELS OF MIR-23A AND BCR/ABL WERE ASSESSED IN 42 NEWLY DIAGNOSED CML PATIENTS, 37 CML PATIENTS IN FIRST COMPLETE REMISSION AND 25 HEALTHY CONTROLS. QUANTITATIVE REAL-TIME PCR, WESTERN BLOT ANALYSIS AND COLONY FORMATION ASSAY WERE USED TO EVALUATE CHANGES INDUCED BY OVEREXPRESSION OR INHIBITION OF MIR-23A OR BCR/ABL. MIR-23A MIMIC OR NEGATIVE CONTROL MIMIC WAS TRANSFECTED INTO A CML CELL LINE (K562) AND TWO LUNG CANCER CELL LINES (H157 AND SKMES1) USING LIPOFECTAMINE 2000, AND THE CELLS WERE USED FOR REAL-TIME REVERSE TRANSCRIPTION-PCR (RT-PCR) AND WESTERN BLOT ANALYSIS. WE FOUND THAT THE DOWNREGULATION OF MIR-23A EXPRESSION WAS A FREQUENT EVENT IN BOTH LEUKEMIA CELL LINES AND PRIMARY LEUKEMIC CELLS FROM PATIENTS WITH DE NOVO CML. THE MICROARRAY RESULTS SHOWED THAT MOST OF THE CML PATIENTS EXPRESSED HIGH LEVELS OF BCR/ABL AND LOW LEVELS OF MIR-23A. REAL-TIME RT-PCR AND WESTERN BLOT ANALYSIS SHOWED THAT THE BCR/ABL LEVELS IN MIR-23A-TRANSFECTED CELLS WERE LOWER THAN THOSE IN THE CONTROL GROUPS. ECTOPIC EXPRESSION OF MIR-23A IN K562 CELLS LED TO CELLULAR SENESCENCE. MOREOVER, WHEN K562 CELLS WERE TREATED WITH 5-AZA-2'-DEOXYCYTIDINE, A DNA METHYLATION INHIBITOR, BCR/ABL EXPRESSION WAS UPREGULATED, WHICH INDICATES EPIGENETIC SILENCING OF MIR-23A IN LEUKEMIC CELLS. BCR/ABL AND MIR-23A EXPRESSIONS WERE INVERSELY RELATED TO CML, AND BCR/ABL EXPRESSION WAS REGULATED BY MIR-23A IN LEUKEMIC CELLS. THE EPIGENETIC SILENCING OF MIR-23A LED TO DEREPRESSION OF BCR/ABL EXPRESSION, AND CONSEQUENTLY CONTRIBUTES TO CML DEVELOPMENT AND PROGRESSION.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
16  5319  39 PTEN IS FUNDAMENTAL FOR ELIMINATION OF LEUKEMIA STEM CELLS MEDIATED BY GSK126 TARGETING EZH2 IN CHRONIC MYELOGENOUS LEUKEMIA. PURPOSE: LEUKEMIA STEM CELLS (LSCS) ARE AN IMPORTANT SOURCE OF TYROSINE KINASE INHIBITOR RESISTANCE AND DISEASE RELAPSE IN PATIENTS WITH CHRONIC MYELOGENOUS LEUKEMIA (CML). TARGETING LSCS MAY BE AN ATTRACTIVE STRATEGY TO OVERRIDE THIS THORNY PROBLEM. GIVEN THAT EZH2 WAS OVEREXPRESSED IN PRIMARY CML CD34(+) CELLS, OUR PURPOSE IN THIS STUDY WAS TO EVALUATE THE EFFECTS OF TARGETING EZH2 ON CML LSCS AND CLARIFY ITS UNDERLYING MECHANISM.EXPERIMENTAL DESIGN: HUMAN PRIMARY CML CD34(+) CELLS AND RETROVIRALLY BCR-ABL-DRIVEN CML MOUSE MODELS WERE EMPLOYED TO EVALUATE THE EFFECTS OF SUPPRESSION OF EZH2 BY GSK126- OR EZH2-SPECIFIC SHRNA IN VITRO AND IN VIVO RECRUITMENT OF EZH2 AND H3K27ME3 ON THE PROMOTER OF TUMOR-SUPPRESSOR GENE PTEN IN CML CELLS WAS MEASURED BY CHROMATIN IMMUNOPRECIPITATION ASSAY.RESULTS: OUR RESULTS SHOWED THAT PHARMACOLOGIC INHIBITION OF EZH2 BY GSK126 NOT ONLY ELICITED APOPTOSIS AND RESTRICTED CELL GROWTH IN CML BULK LEUKEMIA CELLS, BUT ALSO DECREASED LSCS IN CML CD34(+) CELLS WHILE SPARING THOSE FROM NORMAL BONE MARROW CD34(+) CELLS. SUPPRESSION OF EZH2 BY GSK126 OR SPECIFIC SHRNA PROLONGED SURVIVAL OF CML MICE AND REDUCED THE NUMBER OF LSCS IN MICE. EZH2 KNOCKDOWN RESULTED IN ELEVATION OF PTEN AND LED TO IMPAIRED RECRUITMENT OF EZH2 AND H3K27ME3 ON THE PROMOTER OF PTEN GENE. THE EFFECT OF EZH2 KNOCKDOWN IN THE CML MICE WAS AT LEAST PARTIALLY REVERSED BY PTEN KNOCKDOWN.CONCLUSIONS: THESE FINDINGS IMPROVE THE UNDERSTANDING OF THE EPIGENETIC REGULATION OF STEMNESS IN CML LSCS AND WARRANT CLINICAL TRIAL OF GSK126 IN REFRACTORY PATIENTS WITH CML. CLIN CANCER RES; 24(1); 145-57. (C)2017 AACR.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
17  2081  52 EPIGENETIC DOWN-REGULATION OF BIM EXPRESSION IS ASSOCIATED WITH REDUCED OPTIMAL RESPONSES TO IMATINIB TREATMENT IN CHRONIC MYELOID LEUKAEMIA. BACKGROUND: EXPRESSION OF THE PRO-APOPTOTIC BCL-2-INTERACTING MEDIATOR (BIM) HAS RECENTLY BEEN IMPLICATED IN IMATINIB-INDUCED APOPTOSIS OF BCR-ABL1(+) CELLS. HOWEVER, THE MECHANISMS INVOLVED IN THE REGULATION OF BIM IN CML AND ITS ROLE IN THE CLINICAL SETTING HAVE NOT BEEN ESTABLISHED. DESIGN AND METHODS: WE ANALYSED THE MRNA EXPRESSION OF BIM IN 100 NEWLY DIAGNOSED PATIENTS WITH CML IN CHRONIC PHASE BY Q-RT-PCR AND THE PROTEIN LEVELS BY WESTERN BLOT ANALYSIS. METHYLATION STATUS WAS ANALYSED BY BISULPHITE GENOMIC SEQUENCING AND MSP. CML CELL LINES WERE TREATED WITH IMATINIB AND 5-AZA-2'-DEOXYCYTIDINE, AND WERE TRANSFECTED WITH TWO DIFFERENT SIRNAS AGAINST BIM AND CELL PROLIFERATION AND APOPTOSIS WERE ANALYSED. RESULTS: WE DEMONSTRATED THAT DOWN-REGULATION OF BIM EXPRESSION WAS PRESENT IN 36% OF THE PATIENTS AND WAS SIGNIFICANTLY ASSOCIATED WITH A LACK OF OPTIMAL RESPONSE TO IMATINIB AS INDICATED BY THE DECREASE IN CYTOGENETIC AND MOLECULAR RESPONSES AT 6, 12 AND 18 MONTHS IN COMPARISON WITH PATIENTS WITH NORMAL BIM EXPRESSION (P<0.05). EXPRESSION OF BIM WAS MEDIATED BY PROMOTER HYPERMETHYLATION AS DEMONSTRATED BY RESTORATION OF BIM EXPRESSION AFTER TREATMENT OF CML CELLS WITH 5-AZA-2'-DEOXYCYTIDINE. USING CML CELL LINES WITH LOW AND NORMAL EXPRESSION OF BIM WE FURTHER DEMONSTRATED THAT THE EXPRESSION OF BIM IS REQUIRED FOR IMATINIB-INDUCED CML APOPTOSIS. CONCLUSION: OUR DATA INDICATE THAT DOWN-REGULATION OF BIM IS EPIGENETICALLY CONTROLLED BY METHYLATION IN A PERCENTAGE OF CML PATIENTS AND HAS AN UNFAVOURABLE PROGNOSTIC IMPACT, AND THAT THE COMBINATION OF IMATINIB WITH A DE-METHYLATING AGENT MAY RESULT IN IMPROVED RESPONSES IN PATIENTS WITH DECREASED EXPRESSION OF BIM.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
18  3371  33 HISTONE MODIFICATION OF PAIN-RELATED GENE EXPRESSION IN SPINAL CORD NEURONS UNDER A PERSISTENT POSTSURGICAL PAIN-LIKE STATE BY ELECTROCAUTERY. CHRONIC POSTSURGICAL PAIN (CPSP) IS A SERIOUS PROBLEM. WE DEVELOPED A MOUSE MODEL OF CPSP INDUCED BY ELECTROCAUTERY AND EXAMINED THE MECHANISM OF CPSP. IN THIS MOUSE MODEL, WHILE BOTH INCISION AND ELECTROCAUTERY EACH PRODUCED ACUTE ALLODYNIA, PERSISTENT ALLODYNIA WAS ONLY OBSERVED AFTER ELECTROCAUTERY. UNDER THESE CONDITIONS, WE FOUND THAT THE MRNA LEVELS OF SMALL PROLINE RICH PROTEIN 1A (SPRR1A) AND ANNEXIN A10 (ANXA10), WHICH ARE THE KEY MODULATORS OF NEUROPATHIC PAIN, IN THE SPINAL CORD WERE MORE POTENTLY AND PERSISTENTLY INCREASED BY ELECTROCAUTERY THAN BY INCISION. FURTHERMORE, THESE GENES WERE OVEREXPRESSED ALMOST EXCLUSIVELY IN CHRONIC POSTSURGICAL PAIN-ACTIVATED NEURONS. THIS EVENT WAS ASSOCIATED WITH DECREASED LEVELS OF TRI-METHYLATED HISTONE H3 AT LYS27 AND INCREASED LEVELS OF ACETYLATED HISTONE H3 AT LYS27 AT THEIR PROMOTER REGIONS. ON THE OTHER HAND, PERSISTENT ALLODYNIA AND OVEREXPRESSION OF SPRR1A AND ANXA10 AFTER ELECTROCAUTERY WERE DRAMATICALLY SUPPRESSED BY SYSTEMIC ADMINISTRATION OF GSK-J4, WHICH IS A SELECTIVE H3K27 DEMETHYLASE INHIBITOR. THESE RESULTS SUGGEST THAT THE EFFECTS OF ELECTROCAUTERY CONTRIBUTE TO CPSP ALONG WITH SYNAPTIC PLASTICITY AND EPIGENETIC MODIFICATION.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
19  3877  46 KDM6A PROMOTES IMATINIB RESISTANCE THROUGH YY1-MEDIATED TRANSCRIPTIONAL UPREGULATION OF TRKA INDEPENDENTLY OF ITS DEMETHYLASE ACTIVITY IN CHRONIC MYELOGENOUS LEUKEMIA. RATIONALE: DESPITE LANDMARK THERAPY OF CHRONIC MYELOGENOUS LEUKEMIA (CML) WITH TYROSINE KINASE INHIBITORS (TKIS), DRUG RESISTANCE REMAINS PROBLEMATIC. CANCER PATHOGENESIS INVOLVES EPIGENETIC DYSREGULATION AND IN PARTICULAR, HISTONE LYSINE DEMETHYLASES (KDMS) HAVE BEEN IMPLICATED IN TKI RESISTANCE. WE SOUGHT TO IDENTIFY KDMS WITH ALTERED EXPRESSION IN CML AND DEFINE THEIR CONTRIBUTION TO IMATINIB RESISTANCE. METHODS: BIOINFORMATICS SCREENING COMPARED KDM EXPRESSION IN CML VERSUS NORMAL BONE MARROW WITH SHRNA KNOCKDOWN AND FLOW CYTOMETRY USED TO MEASURE EFFECTS ON IMATINIB-INDUCED APOPTOSIS IN K562 CELLS. TRANSCRIPTOMIC ANALYSES WERE PERFORMED AGAINST KDM6A CRISPR KNOCKOUT/SHRNA KNOCKDOWN K562 CELLS ALONG WITH GENE RESCUE EXPERIMENTS USING WILDTYPE AND MUTANT DEMETHYLASE-DEAD KDM6A CONSTRUCTS. CO-IMMUNOPRECIPITATION, LUCIFERASE REPORTER AND CHIP WERE EMPLOYED TO ELUCIDATE MECHANISMS OF KDM6A-DEPENDENT RESISTANCE. RESULTS: AMONGST FIVE KDMS UPREGULATED IN CML, ONLY KDM6A DEPLETION SENSITIZED CML CELLS TO IMATINIB-INDUCED APOPTOSIS. RE-INTRODUCTION OF DEMETHYLASE-DEAD KDM6A AS WELL AS WILD-TYPE KDM6A RESTORED IMATINIB RESISTANCE. RNA-SEQ IDENTIFIED NTRK1 GENE DOWNREGULATION AFTER DEPLETION OF KDM6A. MOREOVER, NTRK1 EXPRESSION POSITIVELY CORRELATED WITH KDM6A IN A SUBSET OF CLINICAL CML SAMPLES AND KDM6A KNOCKDOWN IN FRESH CML ISOLATES DECREASED NTRK1 ENCODED PROTEIN (TRKA) EXPRESSION. MECHANISTICALLY, KDM6A WAS RECRUITED TO THE NTRK1 PROMOTER BY THE TRANSCRIPTION FACTOR YY1 WITH SUBSEQUENT TRKA UPREGULATION ACTIVATING DOWN-STREAM SURVIVAL PATHWAYS TO INVOKE IMATINIB RESISTANCE. CONCLUSION: CONTRARY TO ITS REPORTED ROLE AS A TUMOR SUPPRESSOR AND INDEPENDENT OF ITS DEMETHYLASE FUNCTION, KDM6A PROMOTES IMATINIB-RESISTANCE IN CML CELLS. THE IDENTIFICATION OF THE KDM6A/YY1/TRKA AXIS AS A NOVEL IMATINIB-RESISTANCE MECHANISM REPRESENTS AN UNEXPLORED AVENUE TO OVERCOME TKI RESISTANCE IN CML.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
20  4231  49 METHYLATION OF PROTOCADHERIN 10, A NOVEL TUMOR SUPPRESSOR, IS ASSOCIATED WITH POOR PROGNOSIS IN PATIENTS WITH GASTRIC CANCER. BACKGROUND & AIMS: BY USING METHYLATION-SENSITIVE REPRESENTATIONAL DIFFERENCE ANALYSIS, WE IDENTIFIED PROTOCADHERIN 10 (PCDH10), A GENE THAT ENCODES A PROTOCADHERIN AND IS SILENCED IN A TUMOR-SPECIFIC MANNER. WE ANALYZED ITS EPIGENETIC INACTIVATION, BIOLOGICAL EFFECTS, AND PROGNOSTIC SIGNIFICANCE IN GASTRIC CANCER. METHODS: METHYLATION STATUS WAS EVALUATED BY COMBINED BISULFITE RESTRICTION ANALYSIS AND BISULFITE SEQUENCING. THE EFFECTS OF PCDH10 RE-EXPRESSION WERE DETERMINED IN GROWTH, APOPTOSIS, PROLIFERATION, AND INVASION ASSAYS. PCDH10 TARGET GENES WERE IDENTIFIED BY COMPLEMENTARY DNA MICROARRAY ANALYSIS. RESULTS: PCDH10 WAS SILENCED OR DOWN-REGULATED IN 94% (16 OF 17) OF GASTRIC CANCER CELL LINES; EXPRESSION LEVELS WERE RESTORED BY EXPOSURE TO DEMETHYLATING AGENTS. RE-EXPRESSION OF PCDH10 IN MKN45 GASTRIC CANCER CELLS REDUCED COLONY FORMATION IN VITRO AND TUMOR GROWTH IN MICE; IT ALSO INHIBITED CELL PROLIFERATION (P < .01), INDUCED CELL APOPTOSIS (P < .001), AND REPRESSED CELL INVASION (P < .05), UP-REGULATING THE PRO-APOPTOSIS GENES FAS, CASPASE 8, JUN, AND CDKN1A; THE ANTIPROLIFERATION GENE FGFR; AND THE ANTI-INVASION GENE HTATIP2. PCDH10 METHYLATION WAS DETECTED IN 82% (85 OF 104) OF GASTRIC TUMORS COMPARED WITH 37% (38 OF 104) OF PAIRED NONTUMOR TISSUES (P < .0001). IN THE LATTER, PCDH10 METHYLATION WAS HIGHER IN PRECANCEROUS LESIONS (27 OF 45; 60%) THAN IN CHRONIC GASTRITIS SAMPLES (11 OF 59; 19%) (P < .0001). AFTER A MEDIAN FOLLOW-UP PERIOD OF 16.8 MONTHS, MULTIVARIATE ANALYSIS REVEALED THAT PATIENTS WITH PCDH10 METHYLATION IN ADJACENT NONTUMOR AREAS HAD A SIGNIFICANT DECREASE IN OVERALL SURVIVAL. KAPLAN-MEIER SURVIVAL CURVES SHOWED THAT PCDH10 METHYLATION WAS ASSOCIATED SIGNIFICANTLY WITH SHORTENED SURVIVAL IN STAGE I-III GASTRIC CANCER PATIENTS. CONCLUSIONS: PCDH10 IS A GASTRIC TUMOR SUPPRESSOR; ITS METHYLATION AT EARLY STAGES OF GASTRIC CARCINOGENESIS IS AN INDEPENDENT PROGNOSTIC FACTOR.	2009