1  2480 105 EPIGENETIC UPREGULATION OF LARGE-CONDUCTANCE CA2+-ACTIVATED K+ CHANNEL EXPRESSION IN UTERINE VASCULAR ADAPTATION TO PREGNANCY. OUR PREVIOUS STUDY DEMONSTRATED THAT PREGNANCY INCREASED LARGE-CONDUCTANCE CA(2+)-ACTIVATED POTASSIUM CHANNEL BETA1 SUBUNIT (BKBETA1) EXPRESSION AND LARGE-CONDUCTANCE CA(2+)-ACTIVATED POTASSIUM CHANNEL ACTIVITY IN UTERINE ARTERIES, WHICH WERE ABROGATED BY CHRONIC HYPOXIA. THE PRESENT STUDY TESTED THE HYPOTHESIS THAT PROMOTER METHYLATION/DEMETHYLATION IS A KEY MECHANISM IN EPIGENETIC REPROGRAMMING OF BKBETA1 EXPRESSION PATTERNS IN UTERINE ARTERIES. OVINE BKBETA1 PROMOTER OF 2315 BP SPANNING FROM -2211 TO +104 OF THE TRANSCRIPTION START SITE WAS CLONED, AND AN SP1-380 BINDING SITE THAT CONTAINS CPG DINUCLEOTIDE IN ITS CORE BINDING SEQUENCES WAS IDENTIFIED. SITE-DIRECTED DELETION OF THE SP1 SITE SIGNIFICANTLY DECREASED THE BKBETA1 PROMOTER ACTIVITY. ESTROGEN RECEPTOR-ALPHA BOUND TO THE SP1 SITE THROUGH TETHERING TO SP1 AND UPREGULATED THE EXPRESSION OF BKBETA1. THE SP1 BINDING SITE AT BKBETA1 PROMOTER WAS HIGHLY METHYLATED IN UTERINE ARTERIES OF NONPREGNANT SHEEP, AND METHYLATION INHIBITED TRANSCRIPTION FACTOR BINDING AND BKBETA1 PROMOTER ACTIVITY. PREGNANCY CAUSED A SIGNIFICANT DECREASE IN CPG METHYLATION AT THE SP1 BINDING SITE AND INCREASED SP1 BINDING TO THE BKBETA1 PROMOTER AND BKBETA1 MRNA ABUNDANCE. CHRONIC HYPOXIA DURING GESTATION ABROGATED THIS PREGNANCY-INDUCED DEMETHYLATION AND UPREGULATION OF BKBETA1 EXPRESSION. THE RESULTS PROVIDE EVIDENCE OF A NOVEL MECHANISM OF PROMOTER DEMETHYLATION IN PREGNANCY-INDUCED REPROGRAMMING OF LARGE-CONDUCTANCE CA(2+)-ACTIVATED POTASSIUM CHANNEL EXPRESSION AND FUNCTION IN UTERINE ARTERIES AND SUGGEST NEW INSIGHTS OF EPIGENETIC MECHANISMS LINKING GESTATIONAL HYPOXIA TO ABERRANT UTEROPLACENTAL CIRCULATION AND INCREASED RISK OF PREECLAMPSIA.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
2   919  70 CHRONIC HYPOXIA DURING GESTATION CAUSES EPIGENETIC REPRESSION OF THE ESTROGEN RECEPTOR-ALPHA GENE IN OVINE UTERINE ARTERIES VIA HEIGHTENED PROMOTER METHYLATION. ESTROGEN RECEPTOR-ALPHA (ERALPHA) PLAYS A KEY ROLE IN THE ADAPTATION OF INCREASED UTERINE BLOOD FLOW IN PREGNANCY. CHRONIC HYPOXIA IS A COMMON STRESS TO MATERNAL CARDIOVASCULAR HOMEOSTASIS AND CAUSES INCREASED RISK OF PREECLAMPSIA. STUDIES IN PREGNANT SHEEP DEMONSTRATED THAT HYPOXIA DURING GESTATION DOWNREGULATED ERALPHA GENE EXPRESSION IN UTERINE ARTERIES. THE PRESENT STUDY TESTED THE HYPOTHESIS THAT HYPOXIA CAUSES EPIGENETIC REPRESSION OF THE ERALPHA GENE IN UTERINE ARTERIES VIA HEIGHTENED PROMOTER METHYLATION. OVINE ERALPHA PROMOTER OF 2035 BP SPANNING FROM -2000 TO +35 OF THE TRANSCRIPTION START SITE WAS CLONED. NO ESTROGEN OR HYPOXIA-INDUCIBLE FACTOR RESPONSE ELEMENTS WERE FOUND AT THE PROMOTER. TWO TRANSCRIPTION FACTOR BINDING SITES, USF(-15) AND SP1(-520), CONTAINING CPG DINUCLEOTIDES WERE IDENTIFIED, WHICH HAD SIGNIFICANT EFFECTS ON THE PROMOTER ACTIVITY. THE USF ELEMENT BINDS TRANSCRIPTION FACTORS USF1 AND USF2, AND THE SP1 ELEMENT BINDS SP1, AS WELL AS ERALPHA THROUGH SP1. DELETION OF THE SP1 SITE ABROGATED 17BETA-ESTRADIOL-INDUCED INCREASE IN THE PROMOTER ACTIVITY. IN NORMOXIC CONTROL SHEEP, CPG METHYLATION AT THE SP1 BUT NOT THE USF SITE WAS SIGNIFICANTLY DECREASED IN UTERINE ARTERIES OF PREGNANT AS COMPARED WITH NONPREGNANT ANIMALS. IN PREGNANT SHEEP EXPOSED TO LONG-TERM HIGH-ALTITUDE HYPOXIA, CPG METHYLATION AT BOTH SP1 AND USF SITES IN UTERINE ARTERIES WAS SIGNIFICANTLY INCREASED. METHYLATION INHIBITED TRANSCRIPTION FACTOR BINDING AND THE PROMOTER ACTIVITY. THE RESULTS PROVIDE EVIDENCE OF HYPOXIA CAUSING HEIGHTENED PROMOTER METHYLATION AND RESULTANT ERALPHA GENE REPRESSION IN UTERINE ARTERIES AND SUGGEST NEW INSIGHTS OF MOLECULAR MECHANISMS LINKING GESTATIONAL HYPOXIA TO ABERRANT UTEROPLACENTAL CIRCULATION AND INCREASED RISK OF PREECLAMPSIA.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
3   159  28 ABERRANT PROMOTER HYPERMETHYLATION OF MULTIPLE GENES IN GALLBLADDER CARCINOMA AND CHRONIC CHOLECYSTITIS. PURPOSE: ABERRANT METHYLATION OF 5' GENE PROMOTER REGIONS IS AN EPIGENETIC PHENOMENON THAT IS A MAJOR MECHANISM FOR SILENCING OF TUMOR SUPPRESSOR GENES IN MANY CANCER TYPES. THERE IS LIMITED INFORMATION ABOUT THE MOLECULAR CHANGES INVOLVED IN THE PATHOGENESIS OF GALLBLADDER CARCINOMA (GBC), INCLUDING METHYLATION STATUS. EXPERIMENTAL DESIGN: WE INVESTIGATED THE ABERRANT PROMOTER METHYLATION PROFILE OF 24 KNOWN OR SUSPECTED TUMOR SUPPRESSOR GENES IN 50 GBCS AND COMPARED THOSE RESULTS WITH THE FINDINGS IN 25 CHRONIC CHOLECYSTITIS (CC) SPECIMENS WITHOUT CANCER. THE METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION AND COMBINED RESTRICTION ANALYSIS METHODS WERE USED TO DETECT METHYLATION, AND THE RESULTS WERE CONFIRMED BY SEQUENCING OF CLONED POLYMERASE CHAIN REACTION PRODUCTS. RESULTS: IN GBC, GENE METHYLATION FREQUENCIES VARIED FROM 0% TO 80%. TEN GENES DEMONSTRATED RELATIVELY HIGH FREQUENCIES OF ABERRANT METHYLATION: SHP1 (80%), 3-OST-2 (72%), CDH13 (44%), P15INK4B (44%), CDH1 (38%), RUNX3 (32%), APC (30%), RIZ1 (26%), P16INK4A (24%), AND HPP1 (20%). EIGHT GENES (P73, RARBETA2, SOCS-1, DAPK, DCR2, DCR1, HIN1, AND CHFR) SHOWED LOW FREQUENCIES (2-14%) OF METHYLATION, AND NO METHYLATION OF THE REMAINING SIX GENES (TIMP-3, P57, RASSF1A, CRBP1, SYK, AND NORE1) WAS DETECTED. IN CC, METHYLATION WAS DETECTED FOR SEVEN GENES: SHP1 (88%), P15INK4B (28%), 3-OST-2 (12%), CDH1 (12%), CDH13 (8%), DCR2 (4%), AND P16INK4A (4%). SIGNIFICANTLY HIGHER FREQUENCIES OF METHYLATION IN GBC COMPARED WITH CC WERE DETECTED FOR EIGHT GENES (3-OST-2, CDH13, CDH1, RUNX3, APC, RIZ1, P16INK4A, AND HPP1). OF THOSE, FOUR GENES SHOWED FREQUENT METHYLATION (>30%) IN GBCS. THE MEAN METHYLATION INDEX, AN EXPRESSION OF THE AMOUNT OF METHYLATED GENES BY CASE, WAS SIGNIFICANTLY HIGHER IN GBC (0.196 +/- 0.013) COMPARED WITH CC (0.065 +/- 0.008; P < 0.001). CONCLUSIONS: OUR STUDY CONSTITUTES THE MOST COMPREHENSIVE METHYLATION PROFILE REPORT AVAILABLE IN GBC AND DEMONSTRATES THAT THIS NEOPLASM HAS A DISTINCT PATTERN OF ABNORMAL GENE METHYLATION. WHEREAS GALLBLADDERS FROM HEALTHY INDIVIDUAL WERE NOT AVAILABLE, OUR FINDING OF METHYLATION IN CC CASES WITHOUT CANCER SUGGESTS THAT THIS PHENOMENON REPRESENTS AN EARLY EVENT IN THE PATHOGENESIS OF GBC.	2004	
                                                                                                                                                                                                             
4  4810  17 OBESITY--A CHRONIC HEALTH PROBLEM IN CLONED MICE? THE MAJORITY OF CLONED ANIMALS DERIVED BY NUCLEAR TRANSFER DIE DURING GESTATION AND DISPLAY NEONATAL ABNORMALITIES. HOWEVER, BECAUSE OF THE LONG GENERATION INTERVAL OF CLONED ANIMAL SPECIES, THE LONG-TERM CONSEQUENCES OF CLONING ON HEALTH HAVE BEEN DIFFICULT TO ACCESS. RECENT STUDIES IN MICE HAVE PROVIDED EVIDENCE THAT CLONED ANIMALS MIGHT HAVE CHRONIC HEALTH PROBLEMS SUCH AS OBESITY. OBESITY IN CLONED MICE IS LIKELY TO REFLECT EPIGENETIC ABNORMALITIES THAT ARISE PARTLY FROM INADEQUATE NUCLEAR PROGRAMMING; THESE OBESE MICE DISPLAY A UNIQUE PHENOTYPE THAT IS SUGGESTIVE OF A DEFECT OTHER THAN MALFUNCTION OF THE LEPTIN-MELANOCORTIN SYSTEM, WHICH OCCURS IN MOST RODENT MODELS OF OBESITY AND IN HUMAN OBESITY.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
5   150  28 ABERRANT HYPOMETHYLATION OF THE CANCER-TESTIS ANTIGEN PRAME CORRELATES WITH PRAME EXPRESSION IN ACUTE MYELOID LEUKEMIA. PRAME IS A TUMOR-ASSOCIATED ANTIGEN, WHICH BELONGS TO THE FAMILY OF CANCER-TESTIS ANTIGENS (CTA). THE EXPRESSION OF CTA IS MAINLY RESTRICTED TO THE TESTIS AND VARIOUS TUMORS. IN CONTRAST TO OTHER CTA, PRAME EXPRESSION IS ALSO FREQUENTLY DETECTED IN ACUTE AND CHRONIC LEUKEMIAS. DUE TO THIS EXPRESSION PATTERN, PRAME HAS ATTRACTED GREAT INTEREST AS A PROGNOSTIC TUMOR MARKER THAT CAN BE USED FOR THE DETECTION OF MINIMAL RESIDUAL DISEASE AND AS A POTENTIAL TARGET FOR IMMUNOTHERAPY. IN ACUTE MYELOID LEUKEMIA (AML), PRAME EXPRESSION HAS BEEN OBSERVED IN 30-64% OF CASES. TO EVALUATE WHETHER EPIGENETIC MECHANISMS CONTRIBUTE TO PRAME ACTIVATION IN AML, WE STUDIED DNA METHYLATION OF 15 CPG DINUCLEOTIDES WITHIN A CPG-RICH REGION LOCATED IN THE INTRON 1 OF THE PRAME GENE. DNA METHYLATION WAS DETERMINED BY SEQUENCE ANALYSIS OF CLONED PCR PRODUCTS GENERATED FROM BISULFITE-TREATED GENOMIC DNA. METHYLATION PATTERNS WERE CORRELATED WITH PRAME MRNA LEVELS AS DETERMINED BY MICROARRAY ANALYSIS AND REAL-TIME PCR. WE FOUND ALMOST COMPLETE METHYLATION IN MONONUCLEAR BLOOD CELLS FROM TWO HEALTHY DONORS AND IN BONE MARROW CELLS OF FOUR PRAME-NEGATIVE AML PATIENTS. IN CONTRAST, THE DEGREE OF PRAME METHYLATION WAS CLEARLY REDUCED IN FOUR PRAME-POSITIVE AML BONE MARROW SAMPLES. IN PARTICULAR, THESE SAMPLES WERE CHARACTERIZED BY THE PRESENCE OF CLONES, WHICH WERE COMPLETELY DEVOID OF METHYLATION. THE SIGNIFICANT INVERSE CORRELATION BETWEEN THE DEGREE OF METHYLATION AND PRAME EXPRESSION SUGGESTS A CAUSAL ROLE OF DNA METHYLATION IN PRAME REGULATION. SUCH A ROLE IS FURTHER SUPPORTED BY THE OBSERVATION THAT TREATMENT OF PRAME-NEGATIVE CELL LINES U-937 AND THP-1 WITH THE DEMETHYLATING AGENT 5'-AZA-2'DC RESULTED IN A DOSE-RELATED UPREGULATION OF PRAME EXPRESSION.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
6  2355  25 EPIGENETIC REGULATION OF PRAME GENE IN CHRONIC MYELOID LEUKEMIA. TUMOR ASSOCIATED ANTIGENS (TAA) PROVIDE ATTRACTIVE TARGETS FOR CANCER-SPECIFIC IMMUNOTHERAPY. PRAME IS A TAA GENE UP-REGULATED IN ADVANCED PHASES OF CHRONIC MYELOID LEUKEMIA (CML). TO DATE, MOLECULAR MECHANISMS FOR THE EXPRESSION OF PRAME HAVE NEVER BEEN STUDIED. WE FOUND THAT SOME PH'-POSITIVE CELL LINES DID NOT EXPRESS PRAME. THE EXPRESSION OF PRAME WAS RESTORED IN THESE CELL LINES BY TREATMENT WITH 5'-AZA-2'-DEOXYCYTIDINE, SUGGESTING THAT THE EXPRESSION OF PRAME IS MAINLY SUPPRESSED BY HYPERMETHYLATION. BISULFITE SEQUENCING ANALYSIS OF THE CPG SITES OF THE PRAME EXON 2 IN THESE CANCER CELL LINES REVEALED A CLOSE RELATIONSHIP BETWEEN THE METHYLATION STATUS OF THE PRAME GENE AND ITS EXPRESSION. A METHYLATION-SPECIFIC PCR ANALYSIS DEMONSTRATED THAT HYPOMETHYLATION OF PRAME WAS SIGNIFICANTLY MORE FREQUENT IN CML BLAST CRISIS (70%) THAN IN CHRONIC PHASE (36%) (P=0.01) AND WAS CORRELATED WITH HIGH EXPRESSION LEVELS OF PRAME TRANSCRIPTS (P<0.0001). THESE RESULTS SUGGEST THAT HYPOMETHYLATION OF PRAME UP-REGULATES ITS EXPRESSION IN CML AND MIGHT PLAY A SIGNIFICANT ROLE IN THE PROGRESSION OF THE DISEASE.	2007	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
7  1996  25 EPIGENETIC AND GENETIC ALTERATIONS OF THE EDNRB GENE IN NASOPHARYNGEAL CARCINOMA. BACKGROUND: LOSS OF HETEROZYGOSITY (LOH) AT 13Q22 IS A COMMON EVENT IN NASOPHARYNGEAL CARCINOMA (NPC). EDNRB GENE LOCATED AT 13Q22 HAS BEEN DEMONSTRATED TO BE HYPERMETHYLATED IN SOME KINDS OF TUMORS. IN THE CURRENT STUDY, WE FOCUSED ON THE EPIGENETIC AND GENETIC ALTERATIONS OF EDNRB IN NPC. METHODS: THE MRNA EXPRESSION OF EDNRB WAS DETECTED BY SEMIQUANTITATIVE RT-PCR AND REAL-TIME QUANTITATIVE PCR IN 49 NPC AND 12 CHRONIC NASOPHARYNGITIS BIOPSIES. THE METHYLATION AND LOH STATUS OF EDNRB WERE EXAMINED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION, MICROSATELLITE PCR AND SEQUENCING. WE ALSO EXAMINED THE MRNA EXPRESSION OF EDNRB IN FOUR NPC CELL LINES AFTER 5-AZA-2'-DEOXYCYTIDINE TREATMENT. RESULTS: EDNRB WAS DOWNREGULATED IN PRIMARY NPC TISSUES AND NPC CELL LINES, AND A RELATIVELY HIGHER METHYLATION LEVEL OF EDNRB WAS FOUND IN NPC BIOPSIES (84%) COMPARED TO THAT IN CHRONIC NASOPHARYNGITIS BIOPSIES (42%). TREATMENT OF NPC CELL LINES WITH 5-AZA-2'-DEOXYCYTIDINE ACTIVATED EDNRB EXPRESSION. LOH OF EDNRB GENE WAS ALSO FOUND AT TWO MICROSATELLITE SITES WITH RATIOS OF 6.25 AND 16.67% IN NPC. CONCLUSION: OUR RESULTS SUGGESTED THAT EDNRB EXPRESSION MAY BE AFFECTED BY ABERRANT PROMOTER METHYLATION AND GENE DELETION AND MAY PLAY A ROLE IN THE DEVELOPMENT OF NPC.	2007	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
8  5452  14 REPROGRAMMING DNA METHYLATION IN THE PREIMPLANTATION STAGE: PEEPING WITH DOLLY'S EYES. OOCYTE CYTOPLASMIC FACTORS CAN REPROGRAMME THE SPERM GENOME DURING FERTILISATION OR THE SOMATIC CELL GENOME DURING CLONING. DIVERSE REPROGRAMMING MACHINERY ACTS SEQUENTIALLY AND INTERDEPENDENTLY ON THE IMPORTED GENOME TO DRIVE IT TO TOTIPOTENCY, BUT THEIR THREE-DIMENSIONAL INTERACTIONS IN THE CYTOPLASM REMAIN UNKNOWN. ABERRANT EPIGENETIC PHENOMENA IN EARLY CLONED EMBRYOS INDICATE THAT PARTS OF THE SOMATIC CELL GENOME ARE UNYIELDING TO REPROGRAMMING FORCES, OWING TO THEIR 'KNOTTY' EPIGENETIC FEATURES. THIS FASTIDIOUS NATURE OF THE DONOR GENOME MIGHT PREVENT COMPLETION OF EPIGENETIC REPROGRAMMING. IT MIGHT ALSO HELP TO EXPLAIN THE CHRONIC DEVELOPMENTAL DEFECTS SEEN IN MANY CLONED EMBRYOS.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
9  1695  29 DYNAMIC ASSOCIATION OF P300 WITH THE PROMOTER OF THE G PROTEIN-COUPLED RAT DELTA OPIOID RECEPTOR GENE DURING NGF-INDUCED NEURONAL DIFFERENTIATION. THE G PROTEIN-COUPLED DELTA OPIOID RECEPTOR (DOR) PLAYS A CRITICAL ROLE IN PAIN CONTROL. EMERGING EVIDENCE SHOWS THAT DOR ALSO PLAYS A ROLE IN NEURONAL DIFFERENTIATION AND SURVIVAL. NERVE GROWTH FACTOR (NGF) IS KNOWN TO BE CRITICAL FOR THE DEVELOPMENT AND MAINTENANCE OF THE CENTRAL AND PERIPHERAL NERVOUS SYSTEMS. OUR PREVIOUS STUDIES HAVE SHOWN THAT SUSTAINED ACTIVATION OF NGF/PI3K/AKT/NF-KAPPAB SIGNALING IS ESSENTIAL FOR NGF-INDUCED DOR GENE EXPRESSION DURING NEURONAL DIFFERENTIATION AND THAT THE EPIGENETIC MODIFICATIONS AT HISTONE 3 LYSINE 9 TEMPORALLY CORRELATE WITH THE DOR GENE TRANSCRIPTION. IN THIS STUDY, WE CLONED THE RAT DOR GENE PROMOTER AND IDENTIFIED AN NGF-RESPONSIVE REGION SIMILAR TO THAT FROM THE MOUSE DOR GENE PROMOTER. WE FURTHER IDENTIFIED P300, A KNOWN NF-KAPPAB BINDING PARTNER WITH INTRINSIC HISTONE ACETYLTRANSFERASE ACTIVITY, TO BE DYNAMICALLY ASSOCIATED WITH THE DOR GENE. WE ALSO FOUND THAT ASSEMBLING OF RNA POLYMERASE II (POL II) AT THE PROMOTER TOOK PLACE BEFORE NGF STIMULATION, INDICATING THAT P300 COULD ONLY INTERACT WITH PREASSEMBLED POL II AT THE PROMOTER AFTER NGF STIMULATION. TAKEN TOGETHER, THESE RESULTS IMPLICATE THAT PREASSEMBLY OF THE POL II PREINITIATION COMPLEX, SUSTAINED ACTIVATION OF PI3K/AKT/NF-KAPPAB SIGNALING, AND DYNAMIC P300 ASSOCIATION AT THE PROMOTERS SEQUENTIALLY IS ONE OF THE MECHANISMS OF INDUCTION OF THE LATE PHASE GENES DURING NGF-INDUCED NEURONAL DIFFERENTIATION.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
10  1613  30 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
11  3627  26 INACTIVATION OF LARS2, LOCATED AT THE COMMONLY DELETED REGION 3P21.3, BY BOTH EPIGENETIC AND GENETIC MECHANISMS IN NASOPHARYNGEAL CARCINOMA. ALLELIC LOSS OF CHROMOSOME 3P, INCLUDING THE 3P21.3 REGION, IS FOUND IN 95-100% OF PRIMARY NASOPHARYNGEAL CARCINOMA (NPC) BIOPSIES, SUGGESTING THAT THIS REGION SHOULD HARBOR SOME TUMOR SUPPRESSOR GENES (TSGS) CLOSELY RELATED TO NPC DEVELOPMENT. SEVERAL TSGS LOCATED AT 3P21.3, SUCH AS RASSF1A, LTF AND BLU, HAVE BEEN DEMONSTRATED TO BE INVOLVED IN NPC DEVELOPMENT. LARS2 (LEUCYL-TRNA SYNTHETASE 2, MITOCHONDRIAL) IS ANOTHER GENE LOCATED IN THE CHROMOSOME 3 COMMON ELIMINATED REGION-1 (C3CER1) AT 3P21.3. IN THIS STUDY, WE FOCUSSED ON THE EPIGENETIC AND GENETIC ALTERATIONS OF LARS2 IN NPC. THE MRNA EXPRESSION OF LARS2 WAS DETECTED IN 36 NPC AND 8 CHRONIC NASOPHARYNGITIS (NP) TISSUES BY SEMI-QUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (RT-PCR) AND REAL-TIME RT-PCR. SUBSEQUENTLY, THE MUTATION, ALLELIC LOSS, AND METHYLATION STATUS OF LARS2 WERE ANALYSED BY POLYMERASE CHAIN REACTION-SINGLE-STRAND CONFORMATION POLYMORPHISM (PCR-SSCP), HOMOZYGOUS DELETION (HD) ANALYSIS AND METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION IN PRIMARY NPC TISSUES. NO EXPRESSION OR DOWNREGULATION OF LARS2 WAS OBSERVED IN 78% OF PRIMARY NPC TISSUES. NO MUTATIONS, ASSESSED BY PCR-SSCP AND DNA SEQUENCING, WERE FOUND IN THE PROMOTER REGION AND EXON 1 OF LARS2 IN NPC TISSUES, WHEREAS HD WAS DETECTED IN 28% OF NPC SPECIMENS AT THE LARS2 LOCUS. IN ADDITION, HYPERMETHYLATION OF LARS2 WAS FOUND IN 64% OF NPC SAMPLES BUT ONLY IN 12.5% OF NP BIOPSIES. OUR DATA INDICATE THAT INACTIVATION OF LARS2 BY BOTH GENETIC AND EPIGENETIC MECHANISMS MAY BE A COMMON AND IMPORTANT EVENT IN THE CARCINOGENESIS OF NPC.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
12   982  39 CHRONIC PRENATAL HYPOXIA INDUCES EPIGENETIC PROGRAMMING OF PKCEPSILON GENE REPRESSION IN RAT HEARTS. RATIONALE: EPIDEMIOLOGICAL STUDIES DEMONSTRATE A CLEAR ASSOCIATION OF ADVERSE INTRAUTERINE ENVIRONMENT WITH AN INCREASED RISK OF ISCHEMIC HEART DISEASE IN ADULTHOOD. HYPOXIA IS A COMMON STRESS TO THE FETUS AND RESULTS IN DECREASED PROTEIN KINASE C EPSILON (PKCEPSILON) EXPRESSION IN THE HEART AND INCREASED CARDIAC VULNERABILITY TO ISCHEMIA AND REPERFUSION INJURY IN ADULT OFFSPRING IN RATS. OBJECTIVES: THE PRESENT STUDY TESTED THE HYPOTHESIS THAT FETAL HYPOXIA-INDUCED METHYLATION OF CYTOSINE-PHOSPHATE-GUANINE DINUCLEOTIDES AT THE PKCEPSILON PROMOTER IS REPRESSIVE AND CONTRIBUTES TO PKCEPSILON GENE REPRESSION IN THE HEART OF ADULT OFFSPRING. METHODS AND RESULTS: HYPOXIC TREATMENT OF PREGNANT RATS FROM DAYS 15 TO 21 OF GESTATION RESULTED IN SIGNIFICANT DECREASES IN PKCEPSILON PROTEIN AND MRNA IN FETAL HEARTS. SIMILAR RESULTS WERE OBTAINED IN EX VIVO HYPOXIC TREATMENT OF ISOLATED FETAL HEARTS AND RAT EMBRYONIC VENTRICULAR MYOCYTE CELL LINE H9C2. INCREASED METHYLATION OF PKCEPSILON PROMOTER AT SP1 BINDING SITES, -346 AND -268, WERE DEMONSTRATED IN BOTH FETAL HEARTS OF MATERNAL HYPOXIA AND H9C2 CELLS TREATED WITH 1% O(2) FOR 24 HOURS. WHEREAS HYPOXIA HAD NO SIGNIFICANT EFFECT ON THE BINDING AFFINITY OF SP1 TO THE UNMETHYLATED SITES IN H9C2 CELLS, HEARTS OF FETUSES AND ADULT OFFSPRING, METHYLATION OF BOTH SP1 SITES REDUCED SP1 BINDING. THE ADDITION OF 5-AZA-2'-DEOXYCYTIDINE BLOCKED THE HYPOXIA-INDUCED INCREASE IN METHYLATION OF BOTH SP1 BINDING SITES AND RESTORED PKCEPSILON MRNA AND PROTEIN TO THE CONTROL LEVELS. IN HEARTS OF BOTH FETUSES AND ADULT OFFSPRING, HYPOXIA-INDUCED METHYLATION OF SP1 SITES WAS SIGNIFICANTLY GREATER IN MALES THAN IN FEMALES, AND DECREASED PKCEPSILON MRNA WAS SEEN ONLY IN MALES. IN FETAL HEARTS, THERE WAS SIGNIFICANTLY HIGHER ABUNDANCE OF ESTROGEN RECEPTOR ALPHA AND BETA ISOFORMS IN FEMALES THAN IN MALES. BOTH ESTROGEN RECEPTOR ALPHA AND BETA INTERACTED WITH THE SP1 BINDING SITES IN THE FETAL HEART, WHICH MAY EXPLAIN THE SEX DIFFERENCES IN SP1 METHYLATION IN THE FETAL HEART. ADDITIONALLY, SELECTIVE ACTIVATION OF PKCEPSILON RESTORED THE HYPOXIA-INDUCED CARDIAC VULNERABILITY TO ISCHEMIC INJURY IN OFFSPRING. CONCLUSIONS: THE FINDINGS DEMONSTRATE A DIRECT EFFECT OF HYPOXIA ON EPIGENETIC MODIFICATION OF DNA METHYLATION AND PROGRAMMING OF CARDIAC PKCEPSILON GENE REPRESSION IN A SEX-DEPENDENT MANNER, LINKING FETAL HYPOXIA AND PATHOPHYSIOLOGICAL CONSEQUENCES IN THE HEARTS OF ADULT OFFSPRING.	2010	

13  6816  28 [EXPRESSION, GENETIC AND EPIGENETIC ALTERATIONS OF LTF GENE IN NASOPHARYNGEAL CARCINOMA CELL LINES]. OBJECTIVE: TO INVESTIGATE THE EXPRESSION OF LTF MRNA IN SEVERAL NASOPHARYNGEAL CANCER (NPC) CELL LINES, AND ANALYZE THE RELATIONSHIP BETWEEN THE GENETIC AND EPIGENETIC CHANGES AND EXPRESSION OF LTF GENE. METHODS: THE EXPRESSION LEVEL OF LTF WAS DETECTED IN NPC CELL LINES HNE1, HNE2, HNE3, CNE1, CNE2, 5-8F, 6-10B CELLS AND TISSUES OF 15 CASES OF CHRONIC NASOPHARYNGITIS BY RT-PCR. THE LTF PROTEIN LEVEL WAS ANALYZED BY WESTERN BLOTTING IN 6-10B CELLS. THEN LOH, MUTATION AND METHYLATION STATUS OF LTF WAS EXAMINED BY MICROSATELLITES ANALYSIS, PCR-SSCP, MSP AND BISULFITE GENOMIC SEQUENCING, RESPECTIVELY. RESULTS: 15 CHRONIC NASOPHARYNGITIS TISSUES SHOWED STABLE LTF EXPRESSION, WHILE THERE WERE WEAK EXPRESSION IN 6-10B CELLS AND ABSENT EXPRESSION IN REMAINING DETECTED NPC CELL LINES. THERE WAS A SIGNIFICANTLY LOWER LTF EXPRESSION IN CHRONIC NASOPHARYNGITIS TISSUES (Z = -3.738, P = 0.000). NO LTF PROTEIN EXPRESSION WAS OBSERVED IN 6-10B CELLS. LOH ANALYSIS DEMONSTRATED THAT ALLELE LOSS OF LTF WASN'T FOUND IN NPC CELL LINES. LTF MUTATION WAS NOTED IN 14.3% (1/7) OF NPC CELL LINES. DNA SEQUENCING CONFIRMED THE MUTATION POINT IN THE PROMOTER REGION (-305 BP TO -50 BP) WAS AT -218 BP (DEL T) OF LTF GENE IN THE HNE1 CELL LINE. METHYLATION OF LTF GENE WAS NOT FOUND IN CHRONIC NASOPHARYNGITIS. HOWEVER, METHYLATION OF LTF PROMOTER WAS DETECTED IN ALL NPC CELL LINES. LTF MRNA EXPRESSION WAS INCREASED IN 5-8F AND 6-10B CELL LINES AFTER TREATMENT WITH 5-AZA-2-DEOXYCYTIDINE. CONCLUSION: THERE IS AN INACTIVATION OF EXPRESSION OF LTF GENE IN THE NPC CELL LINES. ITS MOLECULAR MECHANISM MAY BE RELATED WITH METHYLATION OF PROMOTER REGION AND DELETION MUTATION.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
14  5274  15 PROMOTER METHYLATION OF P16 AND EDNRB GENE IN LEUKEMIA PATIENTS IN TAIWAN. BOTH EPIGENETIC AND GENETIC ALTERNATIONS ARE INVOLVED IN CANCER FORMATION. IN THIS STUDY, WE HAVE IDENTIFIED THE METHYLATION FREQUENCY OF P16 AND ENDOTHELIN RECEPTOR TYPE B (EDNRB) OF 26 LEUKEMIA PATIENTS AND 8 RANDOMLY SELECTED NORMAL BLOOD DONORS IN TAIWAN. PROMOTER METHYLATION OF P16 WAS DETECTED IN 85% OF ACUTE LYMPHOCYTIC LEUKEMIA (ALL), 83% IN ACUTE MYELOID LEUKEMIA (AML) WHEREAS NO METHYLATION WAS DETECTED IN CHRONIC MYELOID LEUKEMIA (CML) IN BLAST CRISIS. HYPERMETHYLATION OF EDNRB WAS OBSERVED IN 92% OF ALL, 75% AML AND 100% IN CML IN BLAST CRISIS. NO ABERRANT METHYLATION OF P16 AND EDNRB WAS FOUND IN 8 NORMAL BLOOD DONORS. TAKEN TOGETHER, ABERRANT METHYLATION OF P16 AND EDNRB WAS HIGHLY PREVALENT IN LEUKEMIA PATIENTS IN TAIWAN.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
15   479  30 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	
                                                                                                                                                           
16  3792  29 INTERLEUKIN-1BETA INCREASES THE RISK OF GASTRIC CANCER THROUGH INDUCTION OF ABERRANT DNA METHYLATION IN A MOUSE MODEL. INTERLEUKIN-1BETA (IL-1BETA) HAS A SIGNIFICANT ROLE IN CHRONIC GASTRIC INFLAMMATION AND MANIFESTATIONS OF GASTRIC DISEASES. THE PRESENT STUDY AIMED TO ELUCIDATE THE SPECIFIC ROLE OF IL-1BETA IN INDUCTION OF DNA METHYLATION USING IL-1 RECEPTOR TYPE 1 KNOCKOUT (IL-1R1(-)/(-)) MICE. IN THE PRESENT STUDY, WILD-TYPE (WT) AND IL-1R1(-)/(-) MICE WERE INJECTED WITH IL-1BETA (5 MICROG/KG/DAY). SERUM LEVELS OF IL-1BETA, INTERLEUKIN-6 (IL-6) AND NITRIC OXIDE (NO) WERE MEASURED BY ENZYME-LINKED IMMUNOSORBENT OR NO ASSAYS. E-CADHERIN (E-CAD) METHYLATION STATUS AND MESSENGER (M)RNA EXPRESSION OF IL-1BETA, IL-6, E-CAD AND INDUCIBLE NITRIC OXIDE SYNTHASE (INOS) WERE ANALYZED. RESULTS FROM THE PRESENT STUDY INDICATED SIGNIFICANTLY HIGHER IL-1BETA MRNA EXPRESSION (P<0.001) IN WT MICE COMPARED WITH IL-1R1(-)/(-) MICE. IL-1BETA AND IL-6 RELEASE WAS SIGNIFICANTLY INCREASED IN TREATED WT MICE COMPARED WITH IL-1R1(-)/(-) MICE AT 1 H, 4 H AND 8 H (ALL P<0.005). IL-1BETA RELEASE WAS ONLY DETECTED IN WT MICE FOLLOWING A SECOND DOSE MEASURED AT DAY 3, WEEK 1 AND WEEK 2 WHEN COMPARED WITH IL-1R1(-)/(-) MICE. PROMOTER METHYLATION OF E-CAD AND A DECREASE IN GENE EXPRESSION WAS OBSERVED IN TREATED WT MICE. MRNA EXPRESSION OF INOS IN WT MICE WAS SIGNIFICANTLY INCREASED AT WEEK 1 COMPARED WITH IL-1R1(-)/(-) MICE (P=0.0411). FURTHERMORE, A SIGNIFICANTLY INCREASED LEVEL OF NO PRODUCTION WAS OBSERVED IN TREATED WT MICE (P<0.005 AT 8 H AND WEEK 1; P<0.001 AT 4 H AND DAY 3) WHEN COMPARED WITH IL-1R1(-)/(-) MICE. THE PRESENT RESULTS INDICATED THAT IL-1BETA WAS ABLE TO DIRECTLY INDUCE DNA METHYLATION, WHICH MAY LINK INFLAMMATION-INDUCED EPIGENETIC CHANGES AND THE DEVELOPMENT OF GASTRIC DISEASES.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
17  5348  26 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
18   411  34 ANALYSIS OF GENOME-WIDE DNA METHYLATION PATTERNS IN OBESITY. OBESITY IS A CHRONIC AND COMPLEX PSYCHOSOMATIC DISEASE THAT IS BECOMING INCREASINGLY PREVALENT WORLDWIDE. THIS STUDY AIMED TO ANALYZE WHOLE METHYLATION PROFILES TO UNCOVER THE EPIGENETIC MECHANISMS ASSOCIATED WITH OBESITY. DNA METHYLATION PROFILES IN BLOOD SAMPLES FROM PATIENTS WITH OBESITY AND NORMAL CONTROLS WERE STUDIED USING THE ILLUMINA 850 K METHYLATION MICROARRAY. THE DIAGNOSTIC VALUE OF THE DIFFERENTIALLY METHYLATED GENES WAS DETERMINED USING RECEIVER OPERATING CHARACTERISTIC (ROC) ANALYSIS. THE EXPRESSION OF SELECTED CANDIDATE GENES WAS VERIFIED USING REVERSE TRANSCRIPTION QUANTITATIVE POLYMERASE CHAIN REACTION (RT-QPCR) AND PYROSEQUENCING. A TOTAL OF 9,371 SIGNIFICANTLY DIFFERENTIALLY METHYLATED SITES (7,974 HYPERMETHYLATED SITES AND 1,397 HYPOMETHYLATED SITES) WERE IDENTIFIED IN 4,571 GENES. A DIFFERENCE IN THE DISTRIBUTION OF DIFFERENTIALLY METHYLATED SITES (HYPERMETHYLATED AND HYPOMETHYLATED) IN BOTH GENE STRUCTURES AND CPG ISLANDS WAS OBSERVED. A TOTAL OF 114 KEY DIFFERENTIALLY METHYLATED SITES WERE IDENTIFIED IN THE CPG ISLANDS. ROC RESULTS INDICATED THAT INHIBIN SUBUNIT BETA B (INHBB), HOMEOBOX A9 (HOXA9), TROPONIN T3 (TNNT3), CYCLIC ADENOSINE MONOPHOSPHATE (CAMP)-RESPONSIVE ELEMENT BINDING PROTEIN (CREB)-REGULATED TRANSCRIPTION COACTIVATOR 1 (CRTC1) AND ZINC FINGER AND BTB DOMAIN-CONTAINING 7 B (ZBTB7B) COULD DISCRIMINATE PATIENTS WITH OBESITY FROM NORMAL CONTROLS. RT-QPCR RESULTS OF CRTC1 AND ZBTB7B WERE CONSISTENT WITH OUR METHYLATION PROFILE RESULTS. THE PYROSEQUENCING RESULTS SHOWED THAT THE METHYLATION LEVELS OF CRTC1 CPG SITES (CPG1 AND CPG2-CG11660071) AND INHBB CPG SITES (CPG2) WERE SIGNIFICANTLY CHANGED IN PATIENTS WITH OBESITY COMPARED WITH NORMAL CONTROLS, WHICH WAS CONSISTENT WITH OUR DNA METHYLATION PROFILE RESULTS. OUR STUDY PROVIDES NEW INSIGHTS INTO THE PATHOLOGICAL MECHANISM OF OBESITY.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
19  3813  32 INTRAUTERINE GROWTH RESTRICTION INHIBITS EXPRESSION OF EUKARYOTIC ELONGATION FACTOR 2 KINASE, A REGULATOR OF PROTEIN TRANSLATION. NUTRIENT DEPRIVATION SUPPRESSES PROTEIN SYNTHESIS BY BLOCKING PEPTIDE ELONGATION. TRANSCRIPTIONAL UPREGULATION AND ACTIVATION OF EUKARYOTIC ELONGATION FACTOR 2 KINASE (EEF2K) BLOCKS PEPTIDE ELONGATION BY PHOSPHORYLATING EUKARYOTIC ELONGATION FACTOR 2. PREVIOUS STUDIES EXAMINING PLACENTAS FROM INTRAUTERINE GROWTH RESTRICTED (IUGR) NEWBORN INFANTS SHOW DECREASED EEF2K EXPRESSION AND ACTIVITY DESPITE CHRONIC NUTRIENT DEPRIVATION. HOWEVER, THE EFFECT OF IUGR ON HEPATIC EEF2K EXPRESSION IN THE FETUS IS UNKNOWN. WE, THEREFORE, EXAMINED THE TRANSCRIPTIONAL REGULATION OF HEPATIC EEF2K GENE EXPRESSION IN A SPRAGUE-DAWLEY RAT MODEL OF IUGR. WE FOUND DECREASED HEPATIC EEF2K MRNA AND PROTEIN LEVELS IN IUGR OFFSPRING AT BIRTH COMPARED WITH CONTROL, CONSISTENT WITH PREVIOUS PLACENTAL OBSERVATIONS. FURTHERMORE, THE CPG ISLAND WITHIN THE EEF2K PROMOTER DEMONSTRATED INCREASED METHYLATION AT A CRITICAL USF 1/2 TRANSCRIPTION FACTOR BINDING SITE. IN VITRO METHYLATION OF THIS BINDING SITE CAUSED NEAR COMPLETE LOSS OF EEF2K PROMOTER ACTIVITY, DESIGNATING THIS PROMOTER AS METHYLATION SENSITIVE. THE EEF2K PROMOTOR IN IUGR OFFSPRING ALSO LOST THE PROTECTIVE HISTONE COVALENT MODIFICATIONS ASSOCIATED WITH UNMETHYLATED CGIS. IN ADDITION, THE +1 NUCLEOSOME WAS DISPLACED 3' AND RNA POLYMERASE LOADING WAS REDUCED AT THE IUGR EEF2K PROMOTER. OUR FINDINGS PROVIDE EVIDENCE TO EXPLAIN WHY IUGR-INDUCED CHRONIC NUTRIENT DEPRIVATION DOES NOT RESULT IN THE UPREGULATION OF EEF2K GENE TRANSCRIPTION.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
20  4014  33 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