1   510 122 ASSOCIATION OF RASGRF1 METHYLATION WITH EPILEPTIC SEIZURES. DNA METHYLATION, ONE OF THE MECHANISMS OF EPIGENETIC REGULATION, HAS BEEN SUGGESTED TO BE RELATED WITH EPILEPSY. RASGRF1 IS A PATERNALLY IMPRINTED GENE AND HAS A DIFFERENTIALLY METHYLATED REGION (DMR) AT THE PROMOTER THAT CAN SILENCE GENE EXPRESSION. WE HAVE PREVIOUSLY OBSERVED THE DOWN-REGULATION OF RASGRF1 IN THE TEMPORAL NEOCORTEX OF EPILEPSY PATIENTS AND IN THE HIPPOCAMPUS OF EPILEPTIC ANIMALS. HERE, WE FURTHER EXPLORED THE DYNAMIC CHANGE (1-DAY ACUTE PERIOD, 10-DAY LATENT PERIOD AND 45-DAY CHRONIC PHASE) OF DNA METHYLATION AND RASGRF1 EXPRESSION AFTER ACUTE EPILEPTIC SEIZURES IN KAINIC ACID (KA)-TREATED MICE, AND WE OBSERVED THE IMPACT OF N-PHTHALYL-L-TRYPTOPHAN (RG108), A DNA METHYLTRANSFERASE (DNMT) INHIBITOR, ON AN ACUTE EPILEPTIC MODEL BY POLYMERASE CHAIN REACTION (PCR), WESTERN BLOTTING, AND BISULFITE SEQUENCING PCR (BSP). THE RESULTS DIRECTLY SHOWED THAT THE METHYLATION OF THE RASGRF1 PROMOTER GRADUALLY INCREASED AND REACHED A MAXIMAL LEVEL AT THE LATENT PERIOD, WITH SUBSEQUENT SUPPRESSION OF RASGRF1 MRNA AND PROTEIN EXPRESSION LEVELS, WHICH REACHED A MINIMUM LEVEL IN THE CHRONIC PHASE. RG108 INHIBITED THE INCREASED METHYLATION OF THE RASGRF1 GENE, WITH SIGNIFICANT INHIBITION OCCURRING AT THE LATENT PERIOD, AND RESTORED RASGRF1 EXPRESSION LEVELS IN THE CHRONIC PHASE. IN ADDITION, WE DEMONSTRATED THAT RG108 COULD SUPPRESS ACUTE EPILEPTIC SEIZURES IN KA-TREATED MICE AND EPILEPTIC DISCHARGES IN 4-AMINOPYRIDINE (4-AP)-TREATED HIPPOCAMPAL SLICES. THESE FINDINGS DEMONSTRATE THAT RASGRF1 IS CLOSELY ASSOCIATED WITH EPILEPSY VIA THE ABERRANT METHYLATION OF RASGRF1, AND REGULATING THE METHYLATION STATUS OF RELEVANT GENES MIGHT BE AN INTRIGUING TOPIC IN FUTURE RESEARCH ON EPILEPSY.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
2  3083  33 GENOME-WIDE SCREEN OF DNA METHYLATION CHANGES INDUCED BY LOW DOSE X-RAY RADIATION IN MICE. EPIGENETIC MECHANISMS PLAY A KEY ROLE IN NON-TARGETED EFFECTS OF RADIATION. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE GLOBAL HYPOMETHYLATION AND PROMOTER HYPERMETHYLATION OF PARTICULAR GENES INDUCED BY LOW DOSE RADIATION (LDR). THIRTY MALE BALB/C MICE WERE DIVIDED INTO 3 GROUPS: CONTROL, ACUTELY EXPOSED (0.5 GY X-RAYS), AND CHRONIC EXPOSURE FOR 10 DAYS (0.05GY/DX10D). HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) AND MEDIP-QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR) WERE USED TO STUDY METHYLATION PROFILES. DNMT1 AND MBD2 EXPRESSION WAS DETERMINED BY QPCR AND WESTERN BLOT ASSAYS. METHYLATION AND EXPRESSION OF RAD23B AND DDIT3 WERE DETERMINED BY BISULFATE SEQUENCING PRIMERS (BSP) AND QPCR, RESPECTIVELY. THE RESULTS SHOW THAT LDR INDUCED GENOMIC HYPOMETHYLATION IN BLOOD 2 H POSTIRRADITAION, BUT WAS NOT RETAINED AT 1-MONTH. DNMT1 AND MBD2 WERE DOWNREGULATED IN A TISSUE-SPECIFIC MANNER BUT DID NOT PERSIST. SPECIFIC HYPERMETHYLATION WAS OBSERVED FOR 811 REGIONS IN THE GROUP RECEIVING CHRONIC EXPOSURE, WHICH COVERED ALMOST ALL KEY BIOLOGICAL PROCESSES AS INDICATED BY GO AND KEGG PATHWAY ANALYSIS. EIGHT HYPERMETHYLATED GENES (RAD23B, TDG, CCND1, DDIT3, LLGL1, RASL11A, TBX2, SCL6A15) WERE VERIFIED BY MEDIP-QPCR. AMONG THEM, RAD23B AND DDIT3 GENE DISPLAYED TISSUE-SPECIFIC METHYLATION AND DOWNREGULATION, WHICH PERSISTED FOR 1-MONTH POSTIRRADIATION. THUS, LDR INDUCED GLOBAL HYPOMETHYLATION AND TISSUE-SPECIFIC PROMOTER HYPERMETHYLATION OF PARTICULAR GENES. PROMOTER HYPERMETHYLATION, RATHER THAN GLOBAL HYPOMETHYLATION, WAS RELATIVELY STABLE. DYSREGULATION OF METHYLATION MIGHT BE CORRELATED WITH DOWN-REGULATION OF DNMT1 AND MBD2, BUT MUCH BETTER UNDERSTANDING THE MOLECULAR MECHANISMS INVOLVED IN THIS PROCESS WILL REQUIRE FURTHER STUDY.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
3  5348  64 RASGRF1, A POTENTIAL METHYLATIC MEDIATOR OF ANTI-EPILEPTOGENESIS? EPILEPTOGENESIS, INDUCED BY STATUS EPILEPTICUS (SE), IS A CHRONIC PROCESS, AND INTERVENTION IN THIS PROGRESS MAY PREVENT CHRONIC EPILEPSY. IT HAS BEEN PROPOSED THAT DNA METHYLATION MIGHT BE RELATED WITH EPILEPTOGENESIS. RASGRF1 HAS A DIFFERENTIALLY METHYLATED REGION AT THE PROMOTER WHICH CAN SILENCE GENE EXPRESSION. WE HAVE PREVIOUSLY OBSERVED THE DOWN-REGULATION OF RASGRF1 IN EPILEPSY PATIENTS AND PROVED THAT HYPERMETHYLATION OF RASGRF1 REACHES MAXIMAL LEVEL AT THE LATENT PERIOD IN MICE AFTER KAINATE-INDUCED SE (KA MICE), WITH CORRESPONDING ALTERATION OF RASGRF1 EXPRESSION. IN THE PRESENT STUDY, N-PHTHALYL-L-TRYPTOPHAN (RG108), A DNA METHYLTRANSFERASE INHIBITOR, WAS APPLIED IN KA MICE AT LATENT PHASE AND THE BEHAVIOR, ELECTROENCEPHALOGRAM AND PATHOLOGICAL CHANGES WERE OBSERVED IN CHRONIC PHASE. METHYLATION AND EXPRESSION OF RASGRF1 WERE DETERMINED BY POLYMERASE CHAIN REACTION (PCR), WESTERN BLOTTING, AND BISULFITE SEQUENCING PCR. THE RESULTS SHOWED THAT THE INCIDENCE OF SPONTANEOUS RECURRENT SEIZURES (SRS) WAS SIGNIFICANTLY LOWER IN THE RG108 GROUP THAN THE NORMAL SALINE (NS) GROUP. SUBGROUP ANALYSIS SHOWED SIGNIFICANT HYPERMETHYLATION AND LOWER EXPRESSION OF RASGRF1 IN THE RG108-SRS SUBGROUP AND THE NS-SRS SUBGROUP BUT NOT IN THE RG108-NSRS (NO SRS) SUBGROUP AND THE NS-NSRS SUBGROUP COMPARED WITH THE CONTROL GROUP. NO SIGNIFICANT DIFFERENCE WAS FOUND BETWEEN THE RG108-SRS AND NS-SRS SUBGROUPS. MEANWHILE, HIPPOCAMPAL NEURONAL LOSS WAS OBSERVED IN RG108-SRS AND NS-SRS SUBGROUPS. WE THUS DEMONSTRATED THAT RG108 COULD MODIFY THE PROGRESSION OF EPILEPTOGENESIS AFTER KA INDUCED SE AND PREVENT CHRONIC EPILEPSY. MEANWHILE, HYPERMETHYLATION OF RASGRF1 AFTER KA INDUCED SE COULD BE REVERSED WITH CORRESPONDING CHANGES OF RASGRF1 EXPRESSION. ADDITIONALLY, WE SPECULATED THAT RASGRF1 MIGHT BE A POTENTIAL EPIGENETIC MEDIATOR IN EPILEPTOGENESIS AND CHRONIC EPILEPSY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
4  4245  32 METHYLATION STATUS OF DDIT3 GENE IN CHRONIC MYELOID LEUKEMIA. BACKGROUND: DNA-DAMAGE-INDUCIBLE TRANSCRIPT 3 (DDIT3), A CANDIDATE TUMOR SUPPRESSOR GENE (TSG), HAS BEEN FOUND INVOLVED IN THE REGULATION OF CELLULAR GROWTH AND DIFFERENTIATION. THE EPIGENETIC CHANGES OF TSGS ARE RECENTLY RECOGNIZED AS AN ABNORMAL MECHANISM CONTRIBUTING TO THE DEVELOPMENT OF CHRONIC MYELOID LEUKEMIA (CML). THE AIM OF THIS STUDY WAS TO INVESTIGATE THE METHYLATION STATUS OF DDIT3 GENE IN CML PATIENTS. METHODS: THE METHYLATION STATUS OF DDIT3 PROMOTER WAS DETECTED IN THE BONE MARROW MONONUCLEAR CELLS FROM 53 PATIENTS WITH CML USING METHYLATION-SPECIFIC PCR (MSP). THE EXPRESSION LEVELS OF DDIT3 AND BCR/ABL TRANSCRIPT WERE DETERMINED BY REAL-TIME QUANTITATIVE PCR (RQ-PCR). CLINICAL DATA OF THESE PATIENTS WERE COLLECTED AND ANALYZED. RESULTS: THE ABERRANT METHYLATION OF DDIT3 GENE PROMOTER WAS FOUND IN 35 OF 53 (66%) CML CASES. CORRELATION WAS NOT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND THE AGE, SEX, HEMOGLOBIN CONCENTRATION, PLATELET COUNTS, CHROMOSOMAL ABNORMALITIES, BCR/ABL TRANSCRIPT, AND STAGING OF CML PATIENTS (P > 0.05), BUT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND WBC COUNTS OF CML CASES (R = 0.781, P < 0.001). THE LEVEL OF DDIT3 TRANSCRIPT IN CML PATIENTS WAS SIGNIFICANTLY LOWER THAN THAT IN CONTROLS (MEDIAN 3.28 VS 19.69, P < 0.001), HOWEVER, THERE WAS NO DIFFERENCE IN THE LEVEL OF DDIT3 TRANSCRIPT BETWEEN METHYLATION-POSITIVE CML CASES (0.05-65.32, MEDIAN 2.13) AND METHYLATION- NEGATIVE CML CASES (0.12-126.04, MEDIAN 3.92) (P > 0.05). CONCLUSION: OUR RESULTS DEMONSTRATE THAT ABERRANT METHYLATION OF DDIT3 OCCURS IN CML FREQUENTLY.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
5  4936  31 PATERNAL COMBINED BOTANICALS CONTRIBUTE TO THE PREVENTION OF ESTROGEN RECEPTOR-NEGATIVE MAMMARY CANCER IN TRANSGENIC MICE. BACKGROUND: PARENTAL NUTRITIONAL INTERVENTIONS HAVE CONSIDERABLY AFFECTED GAMETOGENESIS AND EMBRYOGENESIS, LEADING TO THE DIFFERENTIAL SUSCEPTIBILITY OF OFFSPRING TO CHRONIC DISEASES SUCH AS CANCER. MOREOVER, COMBINATORIAL BIOACTIVE DIETS ARE MORE EFFICACIOUS IN AMELIORATING EPIGENETIC ABERRATIONS IN TUMORIGENESIS. OBJECTIVES: WE SOUGHT TO INVESTIGATE THE TRANSGENERATIONAL INFLUENCE AND EPIGENETIC REGULATION OF PATERNAL SULFORAPHANE (SFN)-RICH BROCCOLI SPROUTS (BSP) AND EPIGALLOCATECHIN-3-GALLATE (EGCG)-RICH GREEN TEA POLYPHENOLS (GTPS) CONSUMPTION IN THE PREVENTION OF ESTROGEN RECEPTOR-NEGATIVE [ER(-)] MAMMARY CANCER IN TRANSGENIC MICE. METHODS: HUMAN BREAST CANCER CELLS WERE USED TO DETECT CELL VIABILITY AND EPIGENETIC-RELATED GENE EXPRESSION AFTER TREATMENT WITH EGCG AND/OR SFN. TWENTY-FOUR C3 OR HER2/NEU MALES WERE RANDOMLY ASSIGNED INTO 4 GROUPS AND TREATED WITH CONTROL, 26% BSP (W/W) IN FOOD, 0.5% GTPS (W/V) IN DRINKING WATER OR COMBINED BSP AND GTPS FOR 7 WK BEFORE MATING. TUMOR GROWTH OF NONTREATED FEMALE PUPS WAS MONITORED WEEKLY FOR 19 WK (C3) AND 25 WK (HER2/NEU). TUMOR- AND EPIGENETIC-RELATED PROTEIN EXPRESSION AND ENZYME ACTIVITIES IN MAMMARY TUMORS WERE MEASURED. SPERMS WERE ISOLATED FROM TREATED MALES FOR RNA SEQUENCING AND REDUCED-REPRESENTATION BISULFITE SEQUENCING ANALYSIS. DATA WERE ANALYZED WITH A 2-FACTOR OR 3-FACTOR ANALYSIS OF VARIANCE. RESULTS: EGCG AND SFN INHIBITED BREAST CANCER CELL GROWTH VIA EPIGENETIC REGULATION. COMBINED BSP AND GTPS SYNERGISTICALLY (COMBINATION INDEX < 1) SUPPRESSED TUMOR GROWTH OVER TIME (P < 0.001) IN 2 MOUSE MODELS. KEY TUMOR-RELATED PROTEINS WERE FOUND DIFFERENTIALLY EXPRESSED (P < 0.05) ALONG WITH EPIGENETIC REGULATIONS IN OFFSPRING MAMMARY TUMORS. THE TRANSCRIPTOME PROFILE OF SPERM DERIVED FROM DIETARY-TREATED MALES REVEALED DIFFERENTIALLY EXPRESSED GENES CORRELATED WITH SPERMATOGENESIS AND BREAST CANCER PROGRESSION. DNA METHYLOMES OF THE SPERM AND FURTHER INTEGRATED ANALYSIS WITH TRANSCRIPTOMES INDICATE THAT DNA METHYLATION ALONE MAY NOT CONTRIBUTE TO SUFFICIENT REGULATION IN DIETARY-TREATED SPERM PRONUCLEUS, LEADING TO OFFSPRING TUMOR SUPPRESSION. CONCLUSIONS: COLLECTIVELY, PATERNAL CONSUMPTION OF COMBINED BSP AND GTPS SHOWS POTENTIAL FOR PREVENTING ER(-) MAMMARY CANCER THROUGH TRANSGENERATIONAL EFFECTS. J NUTR 2023;XX:XX-XX.	2023	

6   479  47 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	
                                                        
7  2126  37 EPIGENETIC INACTIVATION OF MIR-34B/C IN ADDITION TO MIR-34A AND DAPK1 IN CHRONIC LYMPHOCYTIC LEUKEMIA. BACKGROUND: TP53 MUTATION/DELETION IS UNCOMMON IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). WE POSTULATED THAT COMPONENTS OF TP53-CENTERED TUMOR SUPPRESSOR NETWORK, MIR-34B/C, IN ADDITION TO DAPK1 AND MIR-34A MIGHT BE INACTIVATED BY DNA HYPERMETHYLATION. MOREOVER, WE TESTED IF MIR-34B/C METHYLATION MIGHT CORRELATE WITH MIR-203 OR MIR-124-1 METHYLATION IN CLL. METHODS: MIR-34B/C, MIR-34A AND DAPK1 METHYLATION WAS STUDIED IN 11 NORMAL CONTROLS, 7 CLL CELL LINES, AND 78 DIAGNOSTIC CLL SAMPLES BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. MEC-1 CELLS WERE TREATED WITH 5-AZA-2'-DEOXYCYTIDINE FOR REVERSAL OF METHYLATION-ASSOCIATED MIRNA SILENCING. TUMOR SUPPRESSOR PROPERTIES OF MIR-34B WERE DEMONSTRATED BY OVER-EXPRESSION OF PRECURSOR MIR-34B IN MEC-1 CELLS. RESULTS: MIR-34B/C PROMOTER WAS UNMETHYLATED IN NORMAL CONTROLS, BUT COMPLETELY METHYLATED IN 4 CLL CELL LINES. MIR-34B/C EXPRESSION WAS INVERSELY CORRELATED WITH MIR-34B/C METHYLATION. DIFFERENT MSP STATUSES OF MIR-34B/C, INCLUDING COMPLETE METHYLATION AND COMPLETE UNMETHYLATION, WERE VERIFIED BY QUANTITATIVE BISULFITE PYROSEQUENCING. 5-AZA-2'-DEOXYCYTIDINE TREATMENT RESULTED IN PROMOTER DEMETHYLATION AND MIR-34B RE-EXPRESSION IN MEC1 CELLS. MOREOVER, OVER-EXPRESSION OF MIR-34B RESULTED IN INHIBITION OF CELLULAR PROLIFERATION AND INCREASED CELL DEATH. IN PRIMARY CLL SAMPLES, MIR-34A, MIR-34B/C AND DAPK1 METHYLATION WAS DETECTED IN 2.6%, 17.9% AND 34.6% OF PATIENTS AT DIAGNOSIS RESPECTIVELY. FURTHERMORE, 39.7%, 3.8% AND 2.6% PATIENTS HAD METHYLATION OF ONE, TWO OR ALL THREE GENES RESPECTIVELY. OVERALL, 46.2% PATIENTS HAD METHYLATION OF AT LEAST ONE OF THESE THREE GENES. BESIDES, MIR-34B/C METHYLATION WAS ASSOCIATED WITH METHYLATION OF MIR-34A (P = 0.03) AND MIR-203 (P = 0.012) IN CLL. CONCLUSIONS: TAKEN TOGETHER, MIR-34B/C IS A TUMOR SUPPRESSOR MIRNA FREQUENTLY METHYLATED, AND HENCE SILENCED IN CLL. TOGETHER WITH DAPK1 METHYLATION, MIR-34B/C METHYLATION IS IMPLICATED IN THE DISRUPTION OF THE TP53-CENTERED TUMOR SUPPRESSOR NETWORK. MOREOVER, THE ASSOCIATION OF MIRNA METHYLATION WARRANTS FURTHER STUDY.	2014	
                                                                                                                                                                                                                                                                   
8  5459  39 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
9  4601  39 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	
                                                                                                                                                                                                                                                                                                                                                                                                                          
10  4899  39 OXIDATIVE STRESS MEDIATES THE APOPTOSIS AND EPIGENETIC MODIFICATION OF THE BCL-2 PROMOTER VIA DNMT1 IN A CIGARETTE SMOKE-INDUCED EMPHYSEMA MODEL. BACKGROUND: EMPHYSEMA IS A CRUCIAL PATHOLOGICAL CHARACTERISTIC OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). OXIDATIVE STRESS, APOPTOSIS AND EPIGENETIC MECHANISMS CONTRIBUTE TO THE PATHOGENESIS OF EMPHYSEMA. HOWEVER, AN ATTEMPT TO ACCURATELY IDENTIFY WHETHER THESE MECHANISMS INTERACT WITH EACH OTHER AND HOW THEY ARE TRIGGERED HAS NEVER BEEN CONDUCTED. METHOD: THE TOTAL REACTIVE OXYGEN SPECIES (ROS) LEVEL, PULMONARY APOPTOSIS AND B-CELL LYMPHOMA/LEUKEMIA-2 (BCL-2) EXPRESSION, AN APOPTOSIS REGULATOR, WERE DETECTED IN SAMPLES FROM COPD PATIENTS. BISULFITE SEQUENCING PCR (BSP) WAS CONDUCTED TO OBSERVE THE ALTERATIONS IN THE METHYLATION OF THE BCL-2 PROMOTER IN SPECIMENS. THE DYSREGULATION OF DNA METHYLTRANSFERASE ENZYME 1 (DNMT1), A VITAL DNA METHYLTRANSFERASE ENZYME, IN THE LUNGS OF PATIENTS WAS CONFIRMED THROUGH WESTERN BLOTTING. TO FIND OUT INTERACTIONS BETWEEN OXIDATIVE STRESS AND DNA METHYLATION IN EMPHYSEMA, MOUSE MODELS WERE BUILT WITH ANTIOXIDANT TREATMENT AND DNMT1 SILENCING, AND WERE EXAMINED WITH THE PULMONARY APOPTOSIS, BCL-2 AND DNMT1 LEVELS, AND EPIGENETIC ALTERATIONS OF BCL-2. RESULTS: HIGHER ROS LEVELS AND PULMONARY APOPTOSIS WERE OBSERVED IN COPD PATIENTS THAN IN HEALTHY CONTROLS. DOWNREGULATED BCL-2 EXPRESSION WITH INCREASED PROMOTER METHYLATION AND DNMT1 PROTEIN EXPRESSION WAS FOUND IN COPD PATIENTS. ANTIOXIDANT TREATMENT REDUCED THE LEVEL OF ROS, DNMT1 PROTEIN AND EMPHYSEMATOUS PROGRESSION IN THE SMOKING MODELS. FOLLOWING DNMT1 BLOCKADE, SMOKING MODELS SHOWED IMPROVED LUNG FUNCTION, PULMONARY APOPTOSIS, EMPHYSEMATOUS PROGRESSION, AND INCREASED BCL-2 PROTEIN LEVEL WITH LESS PROMOTER METHYLATION THAN EMPHYSEMA MICE. CONCLUSION: CIGARETTE-INDUCED OXIDATIVE STRESS MEDIATES PULMONARY APOPTOSIS AND HYPERMETHYLATION OF THE BCL-2 PROMOTER IN EMPHYSEMA MODELS THROUGH DNMT1.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
11  2842  31 FREQUENT CONCOMITANT EPIGENETIC SILENCING OF SOX1 AND SECRETED FRIZZLED-RELATED PROTEINS (SFRPS) IN HUMAN HEPATOCELLULAR CARCINOMA. BACKGROUND AND AIM: EXCEPT FOR GENETIC MUTATIONS, EPIGENETIC CHANGES ARE ALSO INVOLVED IN THE DEVELOPMENT OF HUMAN CANCERS. RECENTLY, WE HAVE IDENTIFIED SOX1, SRY (SEX DETERMINING REGION Y)-BOX 1, IS HYPERMETHYLATED IN CERVICAL CANCER AND OVARIAN CANCER. THEREFORE, WE INVESTIGATED WHETHER PROMOTER HYPERMETHYLATION OF SOX1 IS COMMON IN HEPATOCELLULAR CARCINOMA (HCC). METHODS: WE USED METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MS-PCR) AND BISULFITE SEQUENCING TO ANALYZE THE METHYALTION LEVEL OF THE SOX1 PROMOTER IN SEVEN HCC CELL LINES, 54 CLINICAL HCCS, 42 CIRRHOTIC LIVERS, 21 LIVERS WITH CHRONIC HEPATITIS, AND 15 CONTROL LIVERS. THEN, WE EMPLOYED QUANTITATIVE MS-PCR (QMSP) TO VALIDATE IN AN INDEPENDENT SET OF SAMPLES (60 PAIRED HCCS AND 30 CONTROL LIVERS). FINALLY, WE USED LUCIFERASE REPORTER AND COLONY FORMATION ASSAY TO CHECK THE EFFECT OF SOX1 IN HCC. RESULTS: PROMOTER METHYLATION OF SOX1 WAS SIGNIFICANTLY FREQUENT IN HCC CELL LINES AND CLINICAL HCCS, CIRRHOTIC LIVERS, BUT NOT IN CONTROL LIVERS (P < 0.0001). THERE IS A SIGNIFICANT CORRELATION BETWEEN DOWNREGULATION OF SOX1 EXPRESSION AND PROMOTER METHYLATION. QMSP RESULTS CONFIRMED THAT PROMOTER HYPERMETHYLATION OF SOX1 IS SIGNIFICANTLY MORE FREQUENT IN HCCS THAN CONTROL LIVERS (P < 0.0001). THE FREQUENCY OF SOX1 METHYLATION IN PATIENTS WITH SECRETED FRIZZLED-RELATED PROTEINS (SFRPS) METHYLATION IS SIGNIFICANTLY HIGHER THAN IN PATIENTS WITHOUT SFRPS METHYLATION (P < 0.0001). FURTHERMORE, ECTOPIC EXPRESSION OF SOX1 COULD SUPPRESS T-CELL FACTOR-DEPENDENT TRANSCRIPTIONAL ACTIVITY AND COLONY FORMATION NUMBER IN HCCS. CONCLUSIONS: CONCOMITANT EPIGENETIC SILENCING OF SOX1 AND SFRPS THROUGH PROMOTER HYPERMETHYLATION IS FREQUENT IN HCCS, AND THIS MIGHT CONTRIBUTE TO ABNORMAL ACTIVATION OF CANONICAL WNT SIGNAL PATHWAY.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
12  1620  31 DNA METHYLTRANSFERASE-MEDIATED TRANSCRIPTIONAL SILENCING IN MALIGNANT GLIOMA: A COMBINED WHOLE-GENOME MICROARRAY AND PROMOTER ARRAY ANALYSIS. EPIGENETIC INACTIVATION OF TUMOR SUPPRESSOR GENES IS A COMMON FEATURE IN HUMAN CANCER. PROMOTER HYPERMETHYLATION AND HISTONE DEACETYLATION ARE REVERSIBLE EPIGENETIC MECHANISMS ASSOCIATED WITH TRANSCRIPTIONAL REGULATION. DNA METHYLTRANSFERASES (DNMT1 AND DNMT3B) REGULATE AND MAINTAIN PROMOTER METHYLATION AND ARE OVEREXPRESSED IN HUMAN CANCER. WE PERFORMED WHOLE-GENOME MICROARRAY ANALYSIS TO IDENTIFY GENES WITH ALTERED EXPRESSION AFTER RNAI-INDUCED SUPPRESSION OF DNMT IN A GLIOBLASTOMA MULTIFORME (GBM) CELL LINE. WE THEN IDENTIFIED GENES WITH BOTH DECREASED EXPRESSION AND EVIDENCE OF PROMOTER CPG ISLAND HYPERMETHYLATION IN GBM TISSUE SAMPLES USING A COMBINED WHOLE-GENOME MICROARRAY TRANSCRIPTOME ANALYSIS IN CONJUNCTION WITH A PROMOTER ARRAY ANALYSIS AFTER DNA IMMUNOPRECIPITATION WITH ANTI-5-METHYLCYTIDINE. DNMT1 AND 3B KNOCKDOWN RESULTED IN THE RESTORED EXPRESSION OF 308 GENES THAT ALSO CONTAINED PROMOTER REGION HYPERMETHYLATION. OF THESE, 43 WERE ALSO FOUND TO BE DOWNREGULATED IN GBM TISSUE SAMPLES. THREE DOWNREGULATED GENES WITH HYPERMETHYLATED PROMOTERS AND RESTORED EXPRESSION IN RESPONSE TO ACUTE DNMT SUPPRESSION WERE ASSAYED FOR METHYLATION CHANGES USING BISULFITE SEQUENCE ANALYSIS OF THE PROMOTER REGION AFTER CHRONIC DNMT SUPPRESSION. RESTORATION OF GENE EXPRESSION WAS NOT ASSOCIATED WITH CHANGES IN PROMOTER REGION METHYLATION, BUT RATHER WITH CHANGES IN HISTONE METHYLATION AND CHROMATIN CONFORMATION. TWO OF THE IDENTIFIED GENES EXHIBITED GROWTH SUPPRESSIVE ACTIVITY IN IN VITRO ASSAYS. COMBINING TARGETED GENETIC MANIPULATIONS WITH COMPREHENSIVE GENOMIC AND EXPRESSION ANALYSES PROVIDES A POTENTIALLY POWERFUL NEW APPROACH FOR IDENTIFYING EPIGENETICALLY REGULATED GENES IN GBM.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
13  1632  36 DNMTS ARE INVOLVED IN TGF-BETA1-INDUCED EPITHELIAL-MESENCHYMAL TRANSITIONS IN AIRWAY EPITHELIAL CELLS. CHRONIC RHINOSINUSITIS (CRS) PATHOGENESIS IS CLOSELY RELATED TO TISSUE REMODELING, INCLUDING EPITHELIAL-MESENCHYMAL TRANSITION (EMT). EPIGENETIC MECHANISMS PLAY KEY ROLES IN EMT. DNA METHYLATION, MEDIATED BY DNA METHYLTRANSFERASES (DNMTS), IS AN EPIGENETIC MARKER THAT IS CRITICAL TO EMT. THE GOAL OF THIS STUDY WAS TO DETERMINE WHETHER DNMTS WERE INVOLVED IN TGF-BETA1-INDUCED EMT AND ELUCIDATE THE UNDERLYING MECHANISMS IN NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION AND DNMT ACTIVITY WERE QUANTIFIED. DNMT EXPRESSION WAS MEASURED USING REAL-TIME PCR (QRT-PCR) IN HUMAN CRS TISSUES. MRNA AND PROTEIN LEVELS OF DNMTS, E-CADHERIN, VIMENTIN, ALPHA-SMA, AND FIBRONECTIN WERE DETERMINED USING RT-PCR AND WESTERN BLOTTING, RESPECTIVELY. DNMT1, DNMT3A, AND DNMT3B GENE EXPRESSION WERE KNOCKED DOWN USING SIRNA TRANSFECTION. MAPK PHOSPHORYLATION AND EMT-RELATED TRANSCRIPTION FACTOR LEVELS WERE DETERMINED USING WESTERN BLOTTING. SIGNALING PATHWAYS WERE ANALYZED USING SPECIFIC INHIBITORS OF MAPK. WE DEMONSTRATED THESE DATA IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION, DNMT ACTIVITY, AND DNMT EXPRESSION INCREASED IN CRS TISSUES. DNMT EXPRESSION WAS POSITIVELY CORRELATED WITH LUND-MCKAY CT SCORES. TGF-BETA1 DOSE-DEPENDENTLY INDUCED DNMT EXPRESSION. FURTHER, 5-AZA INHIBITED TGF-BETA1-INDUCED DNMT, SNAIL, AND SLUG EXPRESSION RELATED TO EMT, AS WELL AS P38 AND JNK PHOSPHORYLATION IN A549 CELLS AND TGF-BETA1-INDUCED DNMT EXPRESSION AND EMT IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. TGF-BETA1-INDUCED DNMT EXPRESSION LEADS TO DNA METHYLATION AND EMT VIA P38, JNK, SNAIL, AND SLUG SIGNALING PATHWAYS. INHIBITION OF DNMT SUPPRESSED THE EMT PROCESS AND THEREFORE IS POTENTIALLY A CRS THERAPEUTIC STRATEGY.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
14  1613  33 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
15   912  23 CHRONIC EXPOSURE TO WATER POLLUTANT TRICHLOROETHYLENE INCREASED EPIGENETIC DRIFT IN CD4(+) T CELLS. AIM: AUTOIMMUNE DISEASE AND CD4(+) T-CELL ALTERATIONS ARE INDUCED IN MICE EXPOSED TO THE WATER POLLUTANT TRICHLOROETHYLENE (TCE). WE EXAMINED HERE WHETHER TCE ALTERED GENE-SPECIFIC DNA METHYLATION IN CD4(+) T CELLS AS A POSSIBLE MECHANISM OF IMMUNOTOXICITY. MATERIALS & METHODS: NAIVE AND EFFECTOR/MEMORY CD4(+) T CELLS FROM MICE EXPOSED TO TCE (0.5 MG/ML IN DRINKING WATER) FOR 40 WEEKS WERE EXAMINED BY BISULFITE NEXT-GENERATION DNA SEQUENCING. RESULTS: A PROBABILISTIC MODEL CALCULATED FROM MULTIPLE GENES SHOWED THAT TCE DECREASED METHYLATION CONTROL IN CD4(+) T CELLS. DATA FROM INDIVIDUAL GENES FITTED TO A QUADRATIC REGRESSION MODEL SHOWED THAT TCE INCREASED GENE-SPECIFIC METHYLATION VARIANCE IN BOTH CD4 SUBSETS. CONCLUSION: TCE INCREASED EPIGENETIC DRIFT OF SPECIFIC CPG SITES IN CD4(+) T CELLS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
16  3460  31 HYPOMETHYLATION OF THE IL8 PROMOTER IN NASAL EPITHELIAL CELLS OF PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS. BACKGROUND: IL-8 IS AN IMPORTANT CHEMOKINE IMPLICATED IN THE PATHOGENESIS OF CHRONIC RHINOSINUSITIS (CRS), BUT LITTLE IS KNOWN ABOUT EPIGENETIC REGULATION OF IL8 IN THE PATHOGENESIS OF CRS. OBJECTIVE: WE SOUGHT TO INVESTIGATE THE RELATIONSHIP BETWEEN THE DNA METHYLATION LEVEL IN THE IL8 PROXIMAL PROMOTER AND CRS IN HAN CHINESE SUBJECTS. METHODS: PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS (CRSWNP; N = 187), PATIENTS WITH CHRONIC RHINOSINUSITIS WITHOUT NASAL POLYPS (CRSSNP; N = 89), AND CONTROL SUBJECTS (N = 57) WERE ENROLLED IN 2 INDEPENDENT COHORTS. PURIFIED HUMAN NASAL EPITHELIAL CELLS FROM EACH PARTICIPANT WERE ASSESSED FOR PERCENTAGE DNA METHYLATION OF CPG SITES IN THE IL8 PROXIMAL PROMOTER BY USING BISULFITE PYROSEQUENCING AND FOR FUNCTIONAL ASPECTS OF METHYLATION STATUS BY USING IN VITRO ASSAYS. RESULTS: DNA METHYLATION OF CPG SITES 1, 2, AND 3, RESPECTIVELY, IN THE IL8 PROXIMAL PROMOTER WAS SIGNIFICANTLY DECREASED IN HUMAN NASAL EPITHELIAL CELLS OF PATIENTS WITH CRSWNP COMPARED WITH THAT IN PATIENTS WITH CRSSNP (P < .001) AND CONTROL SUBJECTS (P < .001). PERCENTAGE OF DNA METHYLATION OF THE CPG3 SITE WAS CORRELATED NEGATIVELY WITH BOTH TISSUE EOSINOPHILIC CATIONIC PROTEIN (P < .01) AND MYELOPEROXIDASE (P < .05) LEVELS. IL-1BETA (P < .001) AND TNF-ALPHA (P < .01) SIGNIFICANTLY INCREASED IL8 EXPRESSION ACCOMPANIED BY A REDUCTION IN METHYLATION AT THE CPG3 SITE (P < .001). ELECTROPHORETIC MOBILITY SHIFT ASSAYS DEMONSTRATED THAT METHYLATION STATUS OF CPG3 CHANGED THE BINDING OF OCTAMER-BINDING TRANSCRIPTION FACTOR 1 AND NUCLEAR FACTOR KAPPAB. CONCLUSION: DECREASED DNA METHYLATION OF PARTICULARLY CPG SITES IN THE IL8 PROXIMAL PROMOTER MIGHT PLAY A ROLE IN THE PATHOGENESIS OF CRSWNP.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
17  3448  29 HYPERMETHYLATION OF THE N-MYC DOWNSTREAM-REGULATED GENE 2 PROMOTER IN PERIPHERAL BLOOD MONONUCLEAR CELLS IS ASSOCIATED WITH LIVER FIBROSIS IN CHRONIC HEPATITIS B. DNA METHYLATION IS A FUNDAMENTAL EPIGENETIC MODIFICATION TO REGULATE GENE EXPRESSION. N-MYC DOWNSTREAM-REGULATED GENE (NDRG) 2 IS A CYTOPLASMIC PROTEIN AND PARTICIPATES IN THE PATHOGENESIS OF LIVER FIBROSIS. IN THIS STUDY, THE MRNA EXPRESSION AND METHYLATION STATUS OF NDRG2 WAS EVALUATED IN PATIENTS WITH CHRONIC HEPATITIS B (CHB). THE STUDY INCLUDED 143 CHB PATIENTS AND 65 NORMAL CONTROLS (NC). THE MRNA EXPRESSION OF NDRG2 IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WAS DETECTED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS WAS DETECTED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. THE NDRG2 MRNA LEVEL WAS LOWER IN THE CHB GROUP THAN IN THE NC GROUP (P < 0.001). METHYLATION FREQUENCY OF THE NDRG2 PROMOTER WAS SIGNIFICANTLY HIGHER IN CHB PATIENTS THAN IN THE NC GROUP (52.44% VS. 26.15%, P < 0.001). IMPORTANTLY, THE RELATIVE EXPRESSION LEVELS OF NDRG2 MRNA WERE SIGNIFICANTLY LOWER IN THE METHYLATED GROUP THAN IN THE UNMETHYLATED GROUP IN BOTH CHB PATIENTS AND NC (P < 0.001). FURTHERMORE, A LOWER MRNA LEVEL AND HYPERMETHYLATION OF NDRG2 WERE ASSOCIATED WITH LIVER FIBROSIS AND INFLAMMATION GRADE IN CHB. THE ASPARTATE AMINOTRANSFERASE-TO-PLATELET RATIO INDEX (APRI) SCORE IS WIDELY USED TO PREDICT LIVER FIBROSIS. THE MRNA EXPRESSION LEVELS AND METHYLATION STATUS OF NDRG2 SHOWED A BETTER SCORE COMPARED TO APRI FOR DISCRIMINATING THE SEVERITY OF LIVER FIBROSIS. IN CONCLUSION, HYPERMETHYLATION OF NDRG2 IN PBMCS WAS CORRELATED WITH DECREASED MRNA EXPRESSION AND WITH LIVER FIBROSIS. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS IS A POTENTIAL NONINVASIVE BIOMARKER TO PREDICT THE SEVERITY OF LIVER FIBROSIS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
18  2326  35 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
19  4014  35 LOW-DOSE CD INDUCES HEPATIC GENE HYPERMETHYLATION, ALONG WITH THE PERSISTENT REDUCTION OF CELL DEATH AND INCREASE OF CELL PROLIFERATION IN RATS AND MICE. BACKGROUND: CADMIUM (CD) IS CLASSIFIED AS A HUMAN CARCINOGEN PROBABLY ASSOCIATED WITH EPIGENETIC CHANGES. DNA METHYLATION IS ONE OF EPIGENETIC MECHANISMS BY WHICH CELLS CONTROL GENE EXPRESSION. THEREFORE, THE PRESENT STUDY GENOME-WIDELY SCREENED THE METHYLATION-ALTERED GENES IN THE LIVER OF RATS PREVIOUSLY EXPOSED TO LOW-DOSE CD. METHODOLOGY PRINCIPAL FINDINGS: RATS WERE EXPOSED TO CD AT 20 NMOL/KG EVERY OTHER DAY FOR 4 WEEKS AND GENE METHYLATION WAS ANALYZED AT THE 48(TH) WEEK WITH METHYLATED DNA IMMUNOPRECIPITATION-CPG ISLAND MICROARRAY. AMONG THE 1629 ALTERED GENES, THERE WERE 675 GENES WHOSE PROMOTER CPG ISLANDS (CGIS) WERE HYPERMETHYLATED, 899 GENES WHOSE PROMOTER CGIS WERE HYPOMETHYLATED, AND 55 GENES WHOSE PROMOTER CGIS WERE MIXED WITH HYPER- AND HYPO-METHYLATION. CASPASE-8 GENE PROMOTER CGIS AND TNF GENE PROMOTER CGIS WERE HYPERMETHYLATED AND HYPOMETHYLATED, RESPECTIVELY, ALONG WITH A LOW APOPTOSIS RATE IN CD-TREATED RAT LIVERS. TO LINK THE ABERRANT METHYLATION OF CASPASE-8 AND TNF GENES TO THE LOW APOPTOSIS INDUCED BY LOW-DOSE CD, MICE WERE GIVEN CHRONIC EXPOSURE TO LOW-DOSE CD WITH AND WITHOUT METHYLATION INHIBITOR (5-AZA-2'-DEOXYCTIDENE, 5-AZA). AT THE 48(TH) WEEK AFTER CD EXPOSURE, LIVERS FROM CD-TREATED MICE DISPLAYED THE INCREASED CASPASE-8 CGI METHYLATION AND DECREASED CASPASE-8 PROTEIN EXPRESSION, ALONG WITH SIGNIFICANT INCREASES IN CELL PROLIFERATION AND OVEREXPRESSION OF TGF-BETA1 AND CYTOKERATIN 8/18 (THE LATTER IS A NEW MARKER OF MOUSE LIVER PRENEOPLASTIC LESIONS), ALL WHICH WERE PREVENTED BY 5-AZA TREATMENT. CONCLUSION/SIGNIFICANCE: THESE RESULTS SUGGEST THAT CD-INDUCED GLOBAL GENE HYPERMETHYLATION, MOST LIKELY CASPASE-8 GENE PROMOTER HYPERMETHYLATION THAT DOWN-REGULATED ITS EXPRESSION, LEADING TO THE DECREASED HEPATIC APOPTOSIS AND INCREASED PRENEOPLASTIC LESIONS.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
20  1433  32 DIFFERENTIAL GENOME-WIDE ARRAY-BASED METHYLATION PROFILES IN PROGNOSTIC SUBSETS OF CHRONIC LYMPHOCYTIC LEUKEMIA. GLOBAL HYPOMETHYLATION AND REGIONAL HYPERMETHYLATION ARE WELL-KNOWN EPIGENETIC FEATURES OF CANCER; HOWEVER, IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), STUDIES ON GENOME-WIDE EPIGENETIC MODIFICATIONS ARE LIMITED. HERE, WE ANALYZED THE GLOBAL METHYLATION PROFILES IN CLL, BY APPLYING HIGH-RESOLUTION METHYLATION MICROARRAYS (27,578 CPG SITES) TO 23 CLL SAMPLES, BELONGING TO THE IMMUNOGLOBULIN HEAVY-CHAIN VARIABLE (IGHV) MUTATED (FAVORABLE) AND IGHV UNMUTATED/IGHV3-21 (POOR-PROGNOSTIC) SUBSETS. OVERALL, RESULTS DEMONSTRATED SIGNIFICANT DIFFERENCES IN METHYLATION PATTERNS BETWEEN THESE SUBGROUPS. SPECIFICALLY, IN IGHV UNMUTATED CLL, WE IDENTIFIED METHYLATION OF 7 KNOWN OR CANDIDATE TUMOR SUPPRESSOR GENES (EG, VHL, ABI3, AND IGSF4) AS WELL AS 8 UNMETHYLATED GENES INVOLVED IN CELL PROLIFERATION AND TUMOR PROGRESSION (EG, ADORA3 AND PRF1 ENHANCING THE NUCLEAR FACTOR-KAPPAB AND MITOGEN-ACTIVATED PROTEIN KINASE PATHWAYS, RESPECTIVELY). IN CONTRAST, THESE LATTER GENES WERE SILENCED BY METHYLATION IN IGHV MUTATED PATIENTS. THE ARRAY DATA WERE VALIDATED FOR SELECTED GENES USING METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION, QUANTITATIVE REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION, AND BISULFITE SEQUENCING. FINALLY, THE SIGNIFICANCE OF DNA METHYLATION IN REGULATING GENE PROMOTERS WAS SHOWN BY REINDUCING 4 METHYLATED TUMOR SUPPRESSOR GENES (EG, VHL AND ABI3) IN IGHV UNMUTATED SAMPLES USING THE METHYL-INHIBITOR 5-AZA-2'-DEOXYCYTIDINE. TAKEN TOGETHER, OUR DATA FOR THE FIRST TIME REVEAL DIFFERENCES IN GLOBAL METHYLATION PROFILES BETWEEN PROGNOSTIC SUBSETS OF CLL, WHICH MAY UNFOLD EPIGENETIC SILENCING MECHANISMS INVOLVED IN CLL PATHOGENESIS.	2010