1 4601 157 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	
                                                                                                                                                                                                                                                                                                                                                           
2 3448  39 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
3 5270  43 PROMOTER DNA METHYLATION FREQUENCY AND CLINICOPATHOLOGICAL ROLE OF MIR-129-2 GENE IN PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA. OBJECTIVES: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS CHARACTERIZED BY THE ACCUMULATION OF APPARENTLY MATURE B-TYPE LYMPHOCYTES IN THE LYMPHOHEMATOPOIETIC ORGANS. METHYLATION IN PROMOTERS OF TUMOR SUPPRESSOR GENES IS ONE OF THE MECHANISMS THAT CAUSES BLOOD MALIGNANCY. IN THIS STUDY, WE EVALUATED THE PROMOTER DNA METHYLATION STATUS OF MIR-129-2 TUMOR SUPPRESSOR GENE AND ITS ASSOCIATION WITH CLINICAL AND LABORATORY PARAMETERS OF PATIENTS WITH CLL. METHODS: WE STUDIED THE PROMOTER DNA METHYLATION FREQUENCY OF THE MIR-129-2 GENE IN 50 PATIENTS WITH CLL AND 50 HEALTHY CONTROLS USING METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION METHODS. STATISTICAL ANALYSIS WAS PERFORMED USING SPSS-18 SOFTWARE, AND A P-VALUE < 0.050 WAS CONSIDERED STATISTICALLY SIGNIFICANT. RESULTS: THE FREQUENCY OF PROMOTER DNA METHYLATION OF THE MIR-129-2 GENE WAS SIGNIFICANTLY HIGHER IN THE CLL GROUP COMPARED WITH CONTROL GROUP (38.0% VS. 0.0%, P < 0.001; CHI(2) = 23.457). THE PROMOTER DNA METHYLATION FREQUENCY OF MIR-129-2 GENE WAS NOT SIGNIFICANTLY DIFFERENT BETWEEN THE TWO SEXES (P = 0.236). A SIGNIFICANT BUT WEAK CORRELATION WAS SEEN BETWEEN THE METHYLATED STATE OF THE MIR-129-2 GENE AND ORGANOMEGALY (P = 0.019, R = 0.330) AS WELL AS HEMOGLOBIN LEVELS (P = 0.020, R = -0.233). HOWEVER, BINARY LOGISTIC REGRESSION ANALYSIS INDICATED ORGANOMEGALY AS THE ONLY CLINICAL BIOMARKER WITH A STATISTICALLY SIGNIFICANT ASSOCIATION WITH THE HYPERMETHYLATED MIR-129-2 GENE STATE (P = 0.046). CONCLUSIONS: THE HIGH FREQUENCY OF PROMOTER DNA METHYLATION OF THE MIR-129-2 GENE IN THE CLL GROUP COMPARED TO THE CONTROL GROUP, AS WELL AS ITS SIGNIFICANT ASSOCIATION WITH ORGANOMEGALY, SUGGESTS THE IMPORTANCE OF THIS EPIGENETIC BIOMARKER IN THE PATHOGENESIS AND PROGNOSIS OF CLL DISEASE.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
4  831  55 CHARACTERIZATION OF TET AND IDH GENE EXPRESSION IN CHRONIC LYMPHOCYTIC LEUKEMIA: COMPARISON WITH NORMAL B CELLS AND PROGNOSTIC SIGNIFICANCE. BACKGROUND: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS THE MOST COMMON HEMATOLOGICAL MALIGNANCY IN WESTERN COUNTRIES, CHARACTERIZED BY A HETEROGENEOUS CLINICAL COURSE. ALTHOUGH GENETIC STUDIES HAVE IDENTIFIED CHROMOSOMAL ABERRATIONS OR SPECIFIC MUTATIONS, EPIGENETIC CHANGES HAVE BEEN POORLY CHARACTERIZED IN CLL. METHODS: WE ASSESSED TEN-ELEVEN TRANSLOCATIONS (TET) 1, 2, AND 3, ISOCITRATE DEHYDROGENASE (IDH) 1, AND 2 MESSENGER RNA (MRNA) EXPRESSION USING REAL-TIME PCR ON PURIFIED LEUKEMIC B CELLS FROM 214 CLL PATIENTS (MEDIAN FOLLOW-UP = 75 MONTHS, RANGE 1-380), NORMAL PERIPHERAL BLOOD B CELLS (N = 20), AND UMBILICAL CORD BLOOD B CELLS (N = 21). THE MICROENVIRONMENT INFLUENCE WAS ASSESSED AFTER 24 H CO-CULTURE OF CLL CELLS WITH BONE MARROW MESENCHYMAL STROMAL CELLS (BMSC). FINALLY, 5-HYDROXYMETHYLCYTOSINE LEVEL (%5-HMC) WAS ASSESSED BY ELISA IN CLL CELLS ALONE OR WITH MICROENVIRONMENT STIMULI. RESULTS: TET 1 AND 3 AND IDH2 WERE DECREASED IN CLL CELLS COMPARED WITH HEALTHY B CELLS (P = 0.0221, 0.0013, <0.0001, RESPECTIVELY), WHILE IDH1 WAS OVEREXPRESSED (P = 0.0037). TET2 AND IDH1 WERE SIGNIFICANTLY CORRELATED WITH TREATMENT-FREE SURVIVAL (TFS); PATIENTS WITH HIGH TET2/IDH1 EXPRESSION HAD A HIGHER MEDIAN TFS (111 MONTHS) THAN PATIENTS WITH LOW EXPRESSION (78 MONTHS, P = 0.0071/0.0123). MOREOVER, TET1 EXPRESSION DECREASED (P = 0.0371), WHILE TET3 AND IDH2 EXPRESSION INCREASED (P = 0.0273/0.0039) IN CO-CULTURES. HOWEVER, %5-HMC WAS NOT CORRELATED WITH CLINICAL DATA AND WAS UNCHANGED FOLLOWING MICROENVIRONMENT STIMULI. CONCLUSIONS: DESPITE A SLIGHT DEREGULATION IN CLL CELLS COMPARED WITH NORMAL B CELLS, WE IDENTIFIED A SIGNIFICANT ASSOCIATION BETWEEN TET/IDH GENE EXPRESSION AND PROGNOSIS, SUGGESTING THAT EPIGENETIC CHANGES COULD POTENTIALLY BE ASSOCIATED WITH DISEASE PROGRESSION. MOREOVER, DESPITE AN IDENTICAL %5-HMC, TET GENE EXPRESSION WAS INFLUENCED BY CONTACT WITH BMSC CONFIRMING THE CRUCIAL ROLE OF THE MICROENVIRONMENT IN CLL PATHOGENESIS.	2016	
                                                                                                                                                                                                                                                                                 
5 3461  48 HYPOMETHYLATION-MEDIATED H19 OVEREXPRESSION INCREASES THE RISK OF DISEASE EVOLUTION THROUGH THE ASSOCIATION WITH BCR-ABL TRANSCRIPT IN CHRONIC MYELOID LEUKEMIA. PREVIOUS STUDY HAS REVEALED THAT H19 EXPRESSION IS REQUIRED FOR EFFICIENT TUMOR GROWTH INDUCED BY BCR-ABL IN CHRONIC MYELOID LEUKEMIA (CML). HEREIN, WE FURTHER DETERMINED H19 EXPRESSION AND ITS CLINICAL IMPLICATION IN PATIENTS WITH CML. H19 EXPRESSION AND METHYLATION WERE DETECTED BY REAL-TIME QUANTITATIVE PCR AND REAL-TIME QUANTITATIVE METHYLATION-SPECIFIC PCR, AND THEN CLINICAL IMPLICATION OF H19 EXPRESSION WAS FURTHER ANALYZED. H19 EXPRESSION WAS SIGNIFICANTLY UP-REGULATED IN CML PATIENTS (P < 0.001). H19 EXPRESSION WITH AN AREA UNDER RECEIVER OPERATING CHARACTERISTIC CURVE VALUE OF 0.824 MIGHT SERVE AS A PROMISING BIOMARKER IN DISTINGUISHING CML PATIENTS FROM CONTROLS. THE PATIENTS WITH HIGH H19 EXPRESSION HAD A TENDENCY OF HIGHER WHITE BLOOD CELLS AND BCR-ABL TRANSCRIPT THAN THOSE WITH LOW H19 EXPRESSION. H19 OVEREXPRESSION OCCURRED WITH THE HIGHER FREQUENCY IN BLAST CRISIS STAGE (11/11, 100%), LOWER IN ACCELERATED PHASE (3/5, 60%), AND CHRONIC PHASE (42/62, 66%) STAGES. MOREOVER, PAIRED PATIENTS DURING DISEASE PROGRESSION WITH INCREASED BCR-ABL TRANSCRIPT ALSO SHOWED A SIGNIFICANT UPREGULATION OF H19 EXPRESSION. MEANWHILE, H19 EXPRESSION WAS DECREASED IN FOLLOW-UP PATIENTS WHO ACHIEVED COMPLETE MOLECULAR REMISSION AFTER TYROSINE KINASE INHIBITORS-BASED THERAPY. EPIGENETIC STUDIES SHOWED THAT H19 DIFFERENTIALLY METHYLATED REGION/IMPRINTING CONTROL REGION (DMR/ICR) WAS HYPOMETHYLATED AND ASSOCIATED WITH H19 EXPRESSION IN CML PATIENTS. MOREOVER, DEMETHYLATION OF H19 DMR/ICR REACTIVATED H19 EXPRESSION IN K562 CELLS. COLLECTIVELY, H19 OVEREXPRESSION, A FREQUENT EVENT IN CML, WAS ASSOCIATED WITH HIGHER BCR-ABL TRANSCRIPT INVOLVING IN DISEASE PROGRESSION. MOREOVER, H19 DMR/ICR HYPOMETHYLATION IN CML MAY BE ONE OF THE MECHANISMS MEDIATING H19 OVEREXPRESSION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                   
6 1433  44 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
7 1968  39 EPIGENETIC ALTERATION OF THE SOCS1 GENE IN CHRONIC MYELOID LEUKAEMIA. THE EXPRESSION OF THE SUPPRESSOR OF CYTOKINE SIGNALLING-1 (SOCS1) PROTEIN IS INDUCED IN RESPONSE TO STIMULATION BY SEVERAL CYTOKINES. THE INDUCED SOCS1 INHIBITS THE SIGNALLING PATHWAY THROUGH THE ASSOCIATION WITH A VARIETY OF TYROSINE KINASE PROTEINS. IN THIS STUDY, THE MUTATION ANALYSES, CPG ISLAND METHYLATION STATUS, AND THE EXPRESSION OF THE SOCS1 GENE IN 112 CHRONIC MYELOID LEUKAEMIA (CML) SAMPLES, FIVE LEUKAEMIA CELL LINES, AND 30 NORMAL CONTROLS WERE ANALYSED. NO GENETIC MUTATIONS OF SOCS1 GENE WERE NOTED IN THE CML SAMPLES. THE SOCS1 GENE WAS HYPERMETHYLATED IN 67% AND 46% OF THE BLASTIC AND CHRONIC PHASE CML SAMPLES RESPECTIVELY (P < 0.0001). HOWEVER, THERE WAS NO METHYLATION OF THE SOCS1 GENE IN NORMAL CONTROLS OR CML IN MOLECULAR REMISSION. THE METHYLATION STATUS OF THE SOCS1 GENE IS CONSISTENT WITH THE RESULTS OF THE REAL-TIME QUANTITATIVE REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION AND IMMUNOCYTOCHEMISTRY STAINING. OUR RESULTS DEMONSTRATE THAT THE SOCS1 GENE SILENCING IS CAUSED BY THE METHYLATION OF CPG ISLANDS IN CML AND IS REVERSED TO AN UNMETHYLATED STATUS IN MOLECULAR REMISSION. AS SOCS1 HAS UNIVERSAL ACTIVITY TO NEGATIVELY REGULATE SEVERAL CYTOKINE SIGNALLING PATHWAYS, THE LOSS OF THE NEGATIVE REGULATION OF CYTOKINE SIGNALLING BY THE SOCS1 MAY PLAY A ROLE IN THE PATHOGENESIS OF CML PROGRESSION.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
8 2847  21 FREQUENT P15 PROMOTER METHYLATION IN TUMOR AND PERIPHERAL BLOOD FROM HEPATOCELLULAR CARCINOMA PATIENTS. WE PROSPECTIVELY ANALYZED P15 METHYLATION PATTERNS IN 25 SURGICALLY RESECTED TUMORS AND 130 PLASMA, SERUM, AND BUFFY COAT SAMPLES FROM HEPATOCELLULAR CARCINOMA (HCC) PATIENTS, CONTROLS WITH CHRONIC HEPATITIS/CIRRHOSIS, AND HEALTHY SUBJECTS. USING METHYLATION-SPECIFIC PCR, WE DEMONSTRATED FOR THE FIRST TIME P15 PROMOTER METHYLATION IN 64% OF TUMORS AND 25% (4 OF 16) OF PATIENTS' PLASMA AND SERUM SAMPLES. CONCURRENT P15 AND P16 METHYLATION WAS SHOWN IN 48% OF TUMORS, AND P15/P16 METHYLATION WAS DETECTED IN THE PLASMA/SERUM OF 92% (11 OF 12) OF PATIENTS. OF NOTE, 75% OF 12 PATIENTS WITH CONCURRENT TUMOR METHYLATION DEVELOPED CLINICAL METASTASIS/RECURRENCE (P = 0.027). IN BUFFY COAT SAMPLES, P15 METHYLATION WAS DETECTED IN ALL EIGHT PATIENTS WITH TUMOR P15 METHYLATION, SUGGESTING THE PRESENCE OF CIRCULATING TUMOR CELLS. NONE OF THE CONTROL SAMPLES WERE METHYLATION POSITIVE. OUR DATA UNDERSCORE THE IMPORTANT ROLE(S) OF P15 AND P16 METHYLATION IN HEPATOCARCINOGENESIS AND TUMOR PROGRESSION. AMONG 92% (23 OF 25) OF PATIENTS WITH TUMOR P15/P16 METHYLATION, CIRCULATING TUMOR DNA AND HCC CELLS WERE DETECTED IN THE PERIPHERAL BLOOD OF 87% (20 OF 23) OF PATIENTS. THE COMBINATION OF THESE EPIGENETIC MARKERS MAY PROVE VALUABLE FOR NONINVASIVE HCC DIAGNOSIS AND DISEASE MONITORING.	2000	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
9 3588  47 IMPACT OF TP53 GENE PROMOTER METHYLATION ON CHRONIC LYMPHOCYTIC LEUKEMIA PATHOGENESIS AND PROGRESSION. BACKGROUND: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS A MALIGNANT LYMPHOID DISORDER THAT RESULTS FROM THE OVERGROWTH OF MATURE-LOOKING LYMPHOID CELLS IN THE BLOOD AND LYMPHATIC TISSUE. VARIOUS CLINICAL PRESENTATIONS HAVE BEEN ATTRIBUTED TO THE DISEASE AS A RESULT OF THE DIFFERENT UNDERLYING GENETIC AND EPIGENETIC ALTERATIONS. THE CURRENT STUDY HAS BEEN INITIATED TO STUDY THE ROLE OF AN EPIGENETIC ALTERATION AFFECTING THE PROMOTER OF THE TP53GENE ON CLL PATHOGENESIS AND PROGRESSION. METHODS: THE CURRENT STUDY INVOLVED 54 NEWLY DIAGNOSED PATIENTS PRESENTING WITH CLL AS WELL AS 30 NORMAL INDIVIDUALS AS CONTROLS. AFTER OBTAINING VERBAL CONSENT, DATA COLLECTION WAS DONE AND THE BLOOD COLLECTED FROM ALL ENROLLED INDIVIDUALS FOR HEMATOLOGICAL INVESTIGATIONS AS WELL AS FOR MOLECULAR CATEGORIZATION OF TP53 METHYLATION STATUS. METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MS-PCR) TECHNIQUE WAS USED TO DEFINE THE METHYLATION STATUS OF THE TP53 GENE PROMOTER THAT ENCOMPASSES DNA EXTRACTION, BISULFITE CONVERSION, CONVENTIONAL PCR AMPLIFICATION, RUNNING ON AGAROSE GEL AND DOCUMENTATION. FINALLY, STATISTICAL ANALYSIS WAS DONE TO ASSESS ANY CORRELATION OF THE TP53 EPIGENETIC ALTERATION TO THE DISEASE ETIOLOGY AND THE PROGRESSION. RESULTS: IN THE CURRENT STUDY, ALL CONTROLS AND 42 OF 54 PATIENTS SHOW UNMETHYLATED TP53 GENE PROMOTER; ON THE OTHER HAND, THE METHYLATED PROMOTER WAS DETECTED AMONG 12 PATIENTS WITH A P-VALUE OF 0.001. TP53 GENE PROMOTER METHYLATION SIGNIFICANTLY LINKED TO REDUCED PLATELET COUNT (P-VALUE OF 0.047) AND ADVANCED STAGE AT PRESENTATION (P-VALUE OF 0.076). NO SIGNIFICANT DIFFERENCES WERE SEEN AMONG BOTH METHYLATED AND UNMETHYLATED TP53 PROMOTERS IN RELATION TO THE AGE OF THE AFFECTED INDIVIDUALS, TOTAL WHITE BLOOD CELL COUNTS AND HEMOGLOBIN LEVEL OF THE AFFECTED INDIVIDUALS. CONCLUSION: THE CURRENT STUDY REVEALED A SIGNIFICANT CORRELATION OF TP53 GENE PROMOTER METHYLATION TO CHRONIC LYMPHOCYTIC LEUKEMIA PATHOGENESIS AND LOWER PLATELET COUNTS.	2019	
                                                                                                                                                                                                                                                                                                           
10 2090  35 EPIGENETIC DYSREGULATION OF THE WNT SIGNALLING PATHWAY IN CHRONIC LYMPHOCYTIC LEUKAEMIA. BACKGROUND: WNT SIGNALLING HAS RECENTLY BEEN IMPLICATED IN THE PATHOGENESIS OF CANCER. METHODS: THIS STUDY INVESTIGATED THE ACTIVITY OF WNT SIGNALLING IN PERIPHERAL BLOOD CHRONIC LYMPHOCYTIC LEUKAEMIA (CLL) LYMPHOCYTES, AND THE METHYLATION STATUS OF SEVEN SOLUBLE WNT ANTAGONIST GENES, INCLUDING WIF1, DKK3, APC, SFRP1, SFRP2, SFRP4 AND SFRP5, BY USING METHYLATION-SPECIFIC PCR IN THE PERIPHERAL BLOOD CLL LYMPHOCYTES AND BONE MARROW SAMPLES OF PATIENTS WITH CLL AT DIAGNOSIS. RESULTS: IN THE PERIPHERAL BLOOD CLL LYMPHOCYTES, CONSTITUTIVE ACTIVATION OF WNT SIGNALLING WAS DETECTED, ASSOCIATED WITH HYPERMETHYLATION OF THE SOLUBLE WNT INHIBITOR GENES. IN THE DIAGNOSTIC CLL MARROW SAMPLES, METHYLATION OF THE SEVEN GENES WAS DETECTED IN UP TO 36.4% OF SAMPLES. MOREOVER, 23 (52.3%) PATIENTS HAD METHYLATION OF AT LEAST ONE OF THE SEVEN GENES, OF WHOM 14 (60.8%) HAD METHYLATION OF TWO OR MORE WNT INHIBITOR GENES. APART FROM AN ASSOCIATION OF ADVANCED AGE WITH DKK3 METHYLATION, THERE WAS NO ASSOCIATION OF GENE HYPERMETHYLATION WITH EITHER CLINICAL CHARACTERISTICS (INCLUDING AGE, GENDER, LYMPHOCYTE COUNT AT DIAGNOSIS, RAI STAGE AND POOR-RISK KARYOTYPE) OR SURVIVAL. CONCLUSION: WNT SIGNALLING IS CONSTITUTIVELY ACTIVATED IN CLL B LYMPHOCYTES IN ASSOCIATION WITH METHYLATION OF MULTIPLE SOLUBLE WNT ANTAGONIST GENES. METHYLATION OF THESE SOLUBLE WNT ANTAGONIST GENES, OCCASIONALLY MULTIPLE GENES, IN PRIMARY CLL MARROW SAMPLES SUGGESTS AN IMPORTANT ROLE IN CLL PATHOGENESIS. MOREOVER, THIS STUDY UNDERSCORED THE IMPORTANCE OF STUDYING METHYLATION OF A PANEL OF, BUT NOT INDIVIDUAL, GENES REGULATING A CELLULAR PATHWAY.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
11 2135  43 EPIGENETIC INACTIVATION OF TUMOR SUPPRESSOR GENES IN SERUM OF PATIENTS WITH CUTANEOUS MELANOMA. SMALL AMOUNTS OF CELL-FREE DNA CIRCULATE IN BOTH HEALTHY AND DISEASED HUMAN BLOOD, WHILE INCREASED CONCENTRATIONS OF DNA ARE PRESENT IN THE SERUM OF CANCER PATIENTS. TUMOR-SPECIFIC MUTATIONS OR EPIGENETIC MODIFICATIONS HAVE PREDOMINANTLY BEEN DETECTED IN TISSUE SPECIMENS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE METHYLATION OF FIVE DIFFERENT GENES INVOLVED IN TUMOR SUPPRESSION AND DNA REPAIR (SUPPRESSORS OF CYTOKINE SIGNALING 1 AND 2 (SOCS1, SOCS2)), RAS-ASSOCIATION DOMAIN FAMILY PROTEIN 1A (RASSF1A), D-TYPE P16(INK4A) CYCLIN-DEPENDENT KINASE INHIBITOR (CDKN), AND O6-METHYLGUANINE DNA-METHYLTRANSFERASE (MGMT)) IN THE SERUM OF 100 PATIENTS USING METHYLATION-SPECIFIC PCR. IN ALL, 41 MELANOMA PATIENTS (STAGE I = 18; STAGE II = 10; STAGE III/IV = 13), 13 HEALTHY CONTROLS WITHOUT NEVI, AND 10 INDIVIDUALS WITH MORE THAN 15 NEVI OF >5 MM IN SIZE WERE INVESTIGATED. FOR COMPARISON, SERA FROM PATIENTS WITH OTHER SKIN TUMORS (NINE BASAL CELL CANCERS, FIVE KAPOSI'S SARCOMA), DIFFERENT METASTASIZED CANCERS (FIVE BREAST CANCERS, FIVE COLON CANCERS), AND SEVERAL CHRONIC INFLAMMATORY DISEASES (N = 12) WERE ALSO ANALYZED. IN ADDITION, WE EXAMINED IF METHYLATION WAS INVOLVED IN SILENCING TRANSCRIPTION OF THESE GENES IN 12 MELANOMA SPECIMENS. SOCS1, SOCS2, RASSF1A, CDKN2A, AND MGMT WERE METHYLATED IN 75, 43, 64, 75, AND 64% OF MELANOMA SAMPLES, RESPECTIVELY. OF THE 41 MELANOMA PATIENTS, 83% HAD ONE HYPERMETHYLATED GENE, WHILE 66, 51, AND 41% HAD TWO, THREE, OR FOUR HYPERMETHYLATED GENES, RESPECTIVELY. ALSO, 20% OF THESE PATIENTS SHOWED HYPERMETHYLATION FOR ALL GENES, WHILE ONLY 17% SHOWED NO METHYLATION. IMPORTANTLY, THE METHYLATION PROFILE OF THE SELECTED GENES FROM MELANOMA PATIENTS WAS DISTINCT FROM THE OTHER ANALYZED TUMORS. TRANSCRIPTION OF SOCS1, SOCS2, CDKN2A, AND RASSF1A GENES WAS SIGNIFICANTLY REDUCED IN FRESH MELANOMA SAMPLES, WHILE MGMT SHOWED A 12-FOLD UPREGULATION AT THE MESSENGER RIBONUCLEIC ACID LEVEL (P < 0.001). OUR FINDINGS SUGGEST THAT EPIGENETIC SILENCING OF THE STUDIED TUMOR SUPPRESSOR GENES IS A COMMON AND PROBABLY IMPORTANT MECHANISM FOR MELANOMA FORMATION. THIS CONVENIENT METHOD USING A SIMPLE BLOOD SAMPLE MAY CONTRIBUTE TO CLASSIFICATION OF MELANOMA AND AWAITS CLINICAL VALIDATION.	2006	
                                                        
12 2759  41 EXPRESSION OF IL-17 AND ITS GENE PROMOTER METHYLATION STATUS ARE ASSOCIATED WITH THE PROGRESSION OF CHRONIC HEPATITIS B VIRUS INFECTION. TO EXPLORE INTERLEUKIN-17 (IL-17) AND ITS EPIGENETIC REGULATION DURING THE PROGRESSION OF CHRONIC HEPATITIS B VIRUS (HBV) INFECTION.A TOTAL OF 162 PATIENTS WITH CHRONIC HBV INFECTION, INCLUDING 75 WITH CHRONIC HEPATITIS B (CHB), 54 WITH HEPATITIS B-ASSOCIATED LIVER CIRRHOSIS AND 33 WITH HEPATITIS B-ASSOCIATED HEPATOCELLULAR CARCINOMA (HBV-HCC), WERE ENROLLED IN THIS STUDY. THIRTY HEALTHY ADULTS OF THE SAME ETHNICITY WERE ENROLLED IN THE CONTROL GROUP. WHOLE VENOUS BLOOD WAS OBTAINED FROM THE PATIENTS AND NORMAL CONTROLS (N = 30). CLINICAL AND LABORATORY PARAMETERS WERE ASSESSED, AND WE PERFORMED ENZYME-LINKED IMMUNOSORBENT ASSAY AND QUANTITATIVE REAL-TIME PCR TO MEASURE THE SERUM LEVELS AND RELATIVE MRNA EXPRESSION OF IL-17, RESPECTIVELY. IL-17 PROMOTER METHYLATION IN PERIPHERAL BLOOD MONONUCLEAR CELLS WAS ASSESSED BY METHYLATION-SPECIFIC PCR. WE ANALYZED THE SERUM AND MRNA LEVELS OF IL-17 AND IL-17 PROMOTER METHYLATION IN THE 4 GROUPS AS WELL AS THE EFFECT OF METHYLATION ON SERUM IL-17 LEVELS. CORRELATIONS BETWEEN THE IL-17 PROMOTER METHYLATION STATUS AND CLINICAL PARAMETERS WERE ANALYZED BY SPEARMAN CORRELATION ANALYSIS.COMPARED TO THE NORMAL CONTROL GROUP, THE PATIENT GROUPS EXHIBITED SIGNIFICANTLY HIGHER SERUM AND RELATIVE MRNA LEVELS OF IL-17. THE METHYLATION DISTRIBUTION AMONG THE PATIENTS WAS SIGNIFICANTLY LOWER THAN THAT AMONG THE NORMAL CONTROLS (P < .05), WITH THE HBV-HCC GROUP SHOWING THE LOWEST IL-17 GENE METHYLATION FREQUENCY. THE AVERAGE IL-17 PROMOTER CG METHYLATION LEVEL WAS NEGATIVELY CORRELATED WITH IL-17 MRNA EXPRESSION (R = -0.39, P = .03), AND NEGATIVE CORRELATIONS BETWEEN IL-17 PROMOTER METHYLATION AND PROTHROMBIN TIME ACTIVITY (R = -0.585, P = .035), ALANINE AMINOTRANSFERASE (R = -0.522, P < .01), ASPARTATE AMINOTRANSFERASE (R = -0.315, P < .05), AND THE MODEL FOR END-STAGE LIVER DISEASE SCORE (R = -0.461, P < .05) WERE OBSERVED. IL-17 SERUM LEVELS IN THE METHYLATED-PROMOTER GROUPS WERE SIGNIFICANTLY LOWER THAN THOSE IN THE UNMETHYLATED-PROMOTER GROUPS.IL-17 EXPRESSION AND PROMOTER METHYLATION WERE ASSOCIATED WITH CHRONIC HBV INFECTION PROGRESSION, ESPECIALLY IN THE HBV-HCC GROUP. THE IL-17 PROMOTER STATUS MAY HELP CLINICIANS INITIATE THE CORRECT TREATMENT STRATEGY AT THE CHB STAGE.	2019	

13 6238  44 THE MALIGNANCY SUPPRESSION ROLE OF MIR-23A BY TARGETING THE BCR/ABL ONCOGENE IN CHROMIC MYELOID LEUKEMIA. THE AIM OF THIS STUDY WAS TO INVESTIGATE THE ROLE AND MECHANISM OF MIR-23A IN THE REGULATION OF BCR/ABL AND TO PROVIDE A NEW PROGNOSTIC BIOMARKER FOR CHRONIC MYELOID LEUKEMIA (CML). THE EXPRESSION LEVELS OF MIR-23A AND BCR/ABL WERE ASSESSED IN 42 NEWLY DIAGNOSED CML PATIENTS, 37 CML PATIENTS IN FIRST COMPLETE REMISSION AND 25 HEALTHY CONTROLS. QUANTITATIVE REAL-TIME PCR, WESTERN BLOT ANALYSIS AND COLONY FORMATION ASSAY WERE USED TO EVALUATE CHANGES INDUCED BY OVEREXPRESSION OR INHIBITION OF MIR-23A OR BCR/ABL. MIR-23A MIMIC OR NEGATIVE CONTROL MIMIC WAS TRANSFECTED INTO A CML CELL LINE (K562) AND TWO LUNG CANCER CELL LINES (H157 AND SKMES1) USING LIPOFECTAMINE 2000, AND THE CELLS WERE USED FOR REAL-TIME REVERSE TRANSCRIPTION-PCR (RT-PCR) AND WESTERN BLOT ANALYSIS. WE FOUND THAT THE DOWNREGULATION OF MIR-23A EXPRESSION WAS A FREQUENT EVENT IN BOTH LEUKEMIA CELL LINES AND PRIMARY LEUKEMIC CELLS FROM PATIENTS WITH DE NOVO CML. THE MICROARRAY RESULTS SHOWED THAT MOST OF THE CML PATIENTS EXPRESSED HIGH LEVELS OF BCR/ABL AND LOW LEVELS OF MIR-23A. REAL-TIME RT-PCR AND WESTERN BLOT ANALYSIS SHOWED THAT THE BCR/ABL LEVELS IN MIR-23A-TRANSFECTED CELLS WERE LOWER THAN THOSE IN THE CONTROL GROUPS. ECTOPIC EXPRESSION OF MIR-23A IN K562 CELLS LED TO CELLULAR SENESCENCE. MOREOVER, WHEN K562 CELLS WERE TREATED WITH 5-AZA-2'-DEOXYCYTIDINE, A DNA METHYLATION INHIBITOR, BCR/ABL EXPRESSION WAS UPREGULATED, WHICH INDICATES EPIGENETIC SILENCING OF MIR-23A IN LEUKEMIC CELLS. BCR/ABL AND MIR-23A EXPRESSIONS WERE INVERSELY RELATED TO CML, AND BCR/ABL EXPRESSION WAS REGULATED BY MIR-23A IN LEUKEMIC CELLS. THE EPIGENETIC SILENCING OF MIR-23A LED TO DEREPRESSION OF BCR/ABL EXPRESSION, AND CONSEQUENTLY CONTRIBUTES TO CML DEVELOPMENT AND PROGRESSION.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
14  156  42 ABERRANT METHYLATION OF THE DEATH-ASSOCIATED PROTEIN KINASE 1 (DAPK1) CPG ISLAND IN CHRONIC MYELOID LEUKEMIA. THE DEATH-ASSOCIATED PROTEIN KINASE 1 (DAPK1) GENE IS A CANDIDATE TUMOR SUPPRESSOR (TSG) AND THE ABNORMAL METHYLATION OF DAPK1 GENE HAS BEEN FOUND IN MANY CARCINOMAS. THE EPIGENETIC CHANGES OF TSGS ARE NOW RECOGNIZED AS A MECHANISM CONTRIBUTING TO THE DEVELOPMENT OF CHRONIC MYELOID LEUKEMIA (CML). TO CLARIFY THE ROLE OF DAPK1 IN CML, WE EXAMINED THE METHYLATION STATUS OF DAPK1 IN 49 PATIENTS WITH CML USING METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. THE ABERRANT METHYLATION OF THE DAPK1 GENE WAS FOUND IN 25 OF 49 (51.0%) CML CASES, NOT IN ALL CONTROLS. NO CORRELATION WAS FOUND BETWEEN DAPK1 GENE METHYLATION AND THE AGE, HEMATOLOGIC PARAMETERS, CHROMOSOMAL ABNORMALITIES, THE TYPES AND LEVELS OF BCR/ABL TRANSCRIPTS OF CML PATIENTS. HOWEVER, CORRELATION COULD BE OBSERVED BETWEEN THE SEX AND THE STATUS OF DAPK1 METHYLATION IN CML PATIENTS (R = 0.374, P = 0.008). FURTHERMORE, THERE WAS A SIGNIFICANT CORRELATION BETWEEN DAPK1 METHYLATION AND THE STAGES OF CML (R = 0.354, P = 0.013). THE CML PATIENTS IN ACCELERATED PHASE (AP) AND BLAST CRISIS (BC) HAD HIGHER FREQUENCY OF DAPK1 METHYLATION THAN THOSE IN CHRONIC PHASE (CP) (75.0% VS. 34.5%) (CHI(2) = 7.776, P = 0.005). IN ONE PATIENT, THE STATUS OF DAPK1 METHYLATION BECAME POSITIVE ON THE TRANSITION FROM CP TO AP AND BC. THESE RESULTS SUGGESTED THAT DAPK1 PROMOTER METHYLATION MIGHT PLAY A SIGNIFICANT ROLE IN THE PROGRESSION OF CML.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
15 1473  39 DISTINCT PATTERNS OF GLOBAL PROMOTER METHYLATION IN EARLY STAGE CHRONIC LYMPHOCYTIC LEUKEMIA. GENOMIC AND EPIGENOMIC STUDIES OF CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) ARE RESHAPING OUR UNDERSTANDING OF THE DISEASE AND HAVE PROVIDED NEW PERSPECTIVES FOR A MORE INDIVIDUALIZED DIAGNOSIS AND NEW POTENTIAL THERAPEUTIC TARGETS. IN THIS STUDY, THE GLOBAL PROMOTER METHYLATION PROFILE WAS DETERMINED IN HIGHLY PURIFIED B-CELLS FROM 37 (BINET STAGE A) CLL PATIENTS, USING HIGH-RESOLUTION METHYLATION MICROARRAYS (27,578 CPG). OVERALL, THE METHYLATION PATTERN CORRELATED WITH THE MAJOR BIOLOGICAL (ZAP-70 AND CD38), AND MOLECULAR (IGHV MUTATION) MARKERS, DISTINGUISHING CLL CASES ACCORDING TO IGHV MUTATIONAL STATUS. CELL ADHESION MOLECULES WERE ENRICHED IN THE SIGNATURE OF UNMUTATED (UM) VERSUS MUTATED (M-) CLL. MOREOVER, IN M-CLL CPG HYPER-METHYLATION IN THREE GENES, INCLUDING SPG20, WAS SIGNIFICANTLY ANTI-CORRELATED WITH THE CORRESPONDING GENE EXPRESSION LEVEL. FINALLY, THE CORRELATION BETWEEN THE METHYLATION PATTERN AND CLINICAL PARAMETERS WAS INVESTIGATED. NOTABLY, OUT OF 42 METHYL-PROBES THAT WERE SIGNIFICANTLY ASSOCIATED WITH PROGRESSION FREE SURVIVAL (PFS), HYPER-METHYLATION OF SPG20 WAS ALSO POSITIVELY ASSOCIATED WITH PFS. THESE DATA SUPPORT THE NOTION THAT EPIGENETIC CHANGES HAVE CLINICAL IMPACT IN CLL AND MAY CONTRIBUTE TO THE IDENTIFICATION OF NOVEL CANDIDATE DISEASE-ASSOCIATED GENES POTENTIALLY USEFUL TO PREDICT THE CLINICAL OUTCOME OF EARLY STAGE CLL PATIENTS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
16 3444  42 HYPERMETHYLATION OF E-CADHERIN IN LEUKEMIA. E-CADHERIN GENE IS OFTEN TERMED A "METASTASIS SUPPRESSOR" GENE BECAUSE THE E-CADHERIN PROTEIN CAN SUPPRESS TUMOR CELL INVASION AND METASTASIS. INACTIVATION OF THE E-CADHERIN GENE OCCURS IN UNDIFFERENTIATED SOLID TUMORS BY BOTH GENETIC AND EPIGENETIC MECHANISMS; HOWEVER, THE ROLE OF E-CADHERIN IN HEMATOLOGIC MALIGNANCIES IS ONLY NOW BEING RECOGNIZED. E-CADHERIN EXPRESSION IS ESSENTIAL FOR ERYTHROBLAST AND NORMOBLAST MATURATION, YET EXPRESSION IS REDUCED OR ABSENT IN LEUKEMIC BLAST CELLS. THIS STUDY EXAMINED THE MESSENGER RNA (MRNA) AND PROTEIN EXPRESSION OF THE E-CADHERIN GENE IN BONE MARROW AND BLOOD SAMPLES FROM NORMAL DONORS AND PATIENTS WITH LEUKEMIA. WE FOUND THAT ALL NORMAL DONOR SAMPLES EXPRESSED E-CADHERIN MRNA, WHEREAS BOTH SAMPLES OF ACUTE MYELOGENOUS LEUKEMIA AND CHRONIC LYMPHOCYTIC LEUKEMIA HAD A SIGNIFICANT REDUCTION OR ABSENCE OF EXPRESSION. HOWEVER, NORMAL BLAST COUNTERPARTS EXPRESSED ONLY A LOW LEVEL OF E-CADHERIN SURFACE PROTEIN. SODIUM BISULPHITE GENOMIC SEQUENCING WAS USED TO FULLY CHARACTERIZE THE METHYLATION PATTERNS OF THE CPG ISLAND ASSOCIATED WITH THE E-CADHERIN GENE PROMOTER IN THOSE SAMPLES WITH MATCHED DNA. ALL OF THE NORMAL CONTROL SAMPLES WERE ESSENTIALLY UNMETHYLATED; HOWEVER, 14 OF 18 (78%) OF THE LEUKEMIA SAMPLES HAD ABNORMAL HYPERMETHYLATION OF THE E-CADHERIN CPG ISLAND. IN FACT BOTH ALLELES OF THE E-CADHERIN GENE WERE OFTEN HYPERMETHYLATED. WE CONCLUDE THE E-CADHERIN GENE IS A COMMON TARGET FOR HYPERMETHYLATION IN HEMATOLOGIC MALIGNANCIES.	2000	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
17  157  36 ABERRANT METHYLATION OF TUMOUR SUPPRESSOR GENE ADAM12 IN CHRONIC LYMPOCYTIC LEUKEMIA PATIENTS: APPLICATION OF METHYLATION SPECIFIC-PCR TECHNIQUE. OBJECTIVE: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS A COMMON LEUKEMIA AMONG CAUCASIANS BUT RARE IN ASIANS POPULATION. WE POSTULATED THAT ABERRANT METHYLATION EITHER HYPERMETHYLATION OR PARTIAL METHYLATION MIGHT BE ONE OF THE SILENCING MECHANISMS THAT INACTIVATES THE TUMOUR SUPPRESSOR GENES IN CLL. THIS STUDY AIMED TO COMPARE THE METHYLATION STATUS OF TUMOUR SUPPRESSOR GENE, ADAM12, AMONG CLL PATIENTS AND NORMAL INDIVIDUALS. WE ALSO EVALUATED THE ASSOCIATION BETWEEN METHYLATION OF ADAM12 AND CLINICAL AND DEMOGRAPHIC CHARACTERISTICS OF THE PARTICIPANTS. METHODS: A TOTAL OF 25 CLL PATIENTS AND 25 NORMAL INDIVIDUALS WERE RECRUITED IN THIS STUDY. THE METHYLATION STATUS OF ADAM12 WAS DETERMINED USING METHYLATION-SPECIFIC PCR (MSP); WHEREAS, DNA SEQUENCING METHOD WAS APPLIED FOR VALIDATION OF THE MSP RESULTS. RESULTS: AMONG CLL PATIENTS, 12 (48%) WERE PARTIALLY METHYLATED AND 13 (52%) WERE UNMETHYLATED. MEANWHILE, 5 (20%) AND 20 (80.6%) OF HEALTHY INDIVIDUALS WERE PARTIALLY METHYLATED AND UNMETHYLATED, RESPECTIVELY. THERE WAS A STATISTICALLY SIGNIFICANT ASSOCIATION BETWEEN THE STATUS OF METHYLATION AT ADAM12 AND THE PRESENCE OF CLL (P=0.037). CONCLUSION: THE ABERRANT METHYLATION OF ADAM12 FOUND IN THIS STUDY USING MSP ASSAY MAY PROVIDE NEW EXPOSURE TO CLL THAT MAY IMPROVE THE GAPS INVOLVED IN GENETIC EPIGENETIC STUDY IN CLL.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
18 5603  35 ROS PROMOTE HYPER-METHYLATION OF NDRG2 PROMOTERS IN A DNMTS-DEPENDENT MANNER: CONTRIBUTES TO THE PROGRESSION OF RENAL FIBROSIS. RENAL FIBROSIS IS THE COMMON HISTOPATHOLOGICAL FEATURE OF CHRONIC KIDNEY DISEASES (CKD), AND THERE IS INCREASING EVIDENCE THAT EPIGENETIC REGULATION IS INVOLVED IN THE OCCURRENCE AND PROGRESSION OF RENAL FIBROSIS. N-MYC DOWNSTREAM-REGULATED GENE 2 (NDRG2) IS SIGNIFICANTLY DOWN-REGULATED IN RENAL FIBROSIS, THE MECHANISM OF WHICH REMAINS UNCLEAR. PREVIOUS STUDIES HAVE CONFIRMED THAT THE INHIBITION OF NDRG2 EXPRESSION IN TUMOR CELLS IS RELATED TO HYPER-METHYLATION, MAINLY REGULATED BY DNA METHYLTRANSFERASES (DNMTS). HEREIN, WE EXPLORED THE EXPRESSION OF NDRG2 AND ITS EPIGENETIC REGULATORY MECHANISM IN RENAL FIBROSIS. THE RESULTS SHOWED THAT THE EXPRESSION OF NDRG2 WAS SIGNIFICANTLY INHIBITED IN VIVO AND IN VITRO, WHILE THE OVEREXPRESSION OF NDRG2 EFFECTIVELY ALLEVIATED RENAL FIBROSIS. MEANWHILE, WE FOUND THAT THE EXPRESSION OF DNMT1/3A/3B WAS SIGNIFICANTLY INCREASED IN HYPOXIA-INDUCED HK2 CELLS AND UNILATERAL URETERAL OBSTRUCTION (UUO) MICE ACCOMPANIED BY HYPER-METHYLATION OF THE NDGR2 PROMOTER. METHYLTRANSFERASE INHIBITOR (5-AZA-DC) CORRECTED THE ABNORMAL EXPRESSION OF DNMT1/3A/3B, REDUCED THE METHYLATION LEVEL OF NDRG2 PROMOTER AND RESTORED THE EXPRESSION OF NDRG2. THE UPSTREAM EVENTS THAT MEDIATE CHANGES IN NDRG2 METHYLATION WERE FURTHER EXPLORED. REACTIVE OXYGEN SPECIES (ROS) ARE IMPORTANT EPIGENETIC REGULATORS AND HAVE BEEN SHOWN TO PLAY A KEY ROLE IN RENAL INJURY DUE TO VARIOUS CAUSES. ACCORDINGLY, WE FURTHER EXPLORED WHETHER ROS COULD INDUCE DNA-EPIGENETIC CHANGES OF THE EXPRESSION OF NDRG2 AND THEN PARTICIPATED IN THE DEVELOPMENT OF RENAL FIBROSIS. OUR RESULTS SHOWED THAT MITOCHONDRIA-TARGETED ANTIOXIDANTS (MITO-TEMPO) COULD REVERSE THE EPIGENETIC INHIBITION OF NDRG2 IN A DNMT-SENSITIVE MANNER, SHOWING STRONG ABILITY OF DNA DEMETHYLATION, EXHIBITING EPIGENETIC REGULATION AND ANTI-FIBROSIS EFFECTS SIMILAR TO 5-AZA-DC. MORE IMPORTANTLY, THE ANTI-FIBROTIC EFFECTS OF 5-AZA-DC AND MITO-TEMPO WERE ELIMINATED IN HK2 CELLS WITH NDRG2 KNOCKDOWN. THESE FINDINGS HIGHLIGHT THAT TARGETING ROS-MEDIATED HYPER-METHYLATION OF NDRG2 PROMOTER IS A POTENTIALLY EFFECTIVE THERAPEUTIC STRATEGY FOR RENAL FIBROSIS, WHICH WILL PROVIDE NEW INSIGHTS INTO THE TREATMENT OF CKD.	2023	
                                                              
19 3071  49 GENOME-WIDE DNA METHYLATION PROFILING INTEGRATED WITH GENE EXPRESSION PROFILING IDENTIFIES PAX9 AS A NOVEL PROGNOSTIC MARKER IN CHRONIC LYMPHOCYTIC LEUKEMIA. BACKGROUND: IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), EPIGENOMIC AND GENOMIC STUDIES HAVE EXPANDED THE EXISTING KNOWLEDGE ABOUT THE DISEASE BIOLOGY AND LED TO THE IDENTIFICATION OF POTENTIAL BIOMARKERS RELEVANT FOR IMPLEMENTATION OF PERSONALIZED MEDICINE. IN THIS STUDY, AN ATTEMPT HAS BEEN MADE TO EXAMINE AND INTEGRATE THE GLOBAL DNA METHYLATION CHANGES WITH GENE EXPRESSION PROFILE AND THEIR IMPACT ON CLINICAL OUTCOME IN EARLY STAGE CLL PATIENTS. RESULTS: THE INTEGRATION OF DNA METHYLATION PROFILE (N = 14) WITH THE GENE EXPRESSION PROFILE (N = 21) REVEALED 142 GENES AS HYPERMETHYLATED-DOWNREGULATED AND; 62 GENES AS HYPOMETHYLATED-UPREGULATED IN EARLY STAGE CLL PATIENTS COMPARED TO CD19+ B-CELLS FROM HEALTHY INDIVIDUALS. THE MRNA EXPRESSION LEVELS OF 17 GENES IDENTIFIED TO BE DIFFERENTIALLY METHYLATED AND/OR DIFFERENTIALLY EXPRESSED WAS FURTHER EXAMINED IN EARLY STAGE CLL PATIENTS (N = 93) BY QUANTITATIVE REAL TIME PCR (RQ-PCR). SIGNIFICANT DIFFERENCES WERE OBSERVED IN THE MRNA EXPRESSION OF MEIS1, PMEPA1, SOX7, SPRY1, CDK6, TBX2, AND SPRY2 GENES IN CLL CELLS AS COMPARED TO B-CELLS FROM HEALTHY INDIVIDUALS. THE ANALYSIS IN THE IGHV MUTATION BASED CATEGORIES (UNMUTATED = 39, MUTATED = 54) REVEALED SIGNIFICANTLY HIGHER MRNA EXPRESSION OF CRY1 AND PAX9 GENES IN THE IGHV UNMUTATED SUBGROUP (P < 0.001). THE RELATIVE RISK OF TREATMENT INITIATION WAS SIGNIFICANTLY HIGHER AMONG PATIENTS WITH HIGH EXPRESSION OF CRY1 (RR = 1.91, P = 0.005) OR PAX9 (RR = 1.87, P = 0.001). HIGH EXPRESSION OF CRY1 (HR: 3.53, P < 0.001) OR PAX9 (HR: 3.14, P < 0.001) GENE WAS SIGNIFICANTLY ASSOCIATED WITH SHORTER TIME TO FIRST TREATMENT. THE HIGH EXPRESSION OF PAX9 GENE (HR: 3.29, 95% CI 1.172-9.272, P = 0.016) WAS ALSO PREDICTIVE OF SHORTER OVERALL SURVIVAL IN CLL. CONCLUSIONS: THE DNA METHYLATION CHANGES ASSOCIATED WITH MRNA EXPRESSION OF CRY1 AND PAX9 GENES ALLOW RISK STRATIFICATION OF EARLY STAGE CLL PATIENTS. THIS COMPREHENSIVE ANALYSIS SUPPORTS THE CONCEPT THAT THE EPIGENETIC CHANGES ALONG WITH THE ALTERED EXPRESSION OF GENES HAVE THE POTENTIAL TO PREDICT CLINICAL OUTCOME IN EARLY STAGE CLL PATIENTS.	2017	
                                                                                                                    
20 3062  39 GENOME-WIDE DNA METHYLATION ANALYSIS REVEALS NOVEL EPIGENETIC CHANGES IN CHRONIC LYMPHOCYTIC LEUKEMIA. WE CONDUCTED A GENOME-WIDE DNA METHYLATION ANALYSIS IN CD19 (+) B-CELLS FROM CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) PATIENTS AND NORMAL CONTROL SAMPLES USING REDUCED REPRESENTATION BISULFITE SEQUENCING (RRBS). THE METHYLATION STATUS OF 1.8-2.3 MILLION CPGS IN THE CLL GENOME WAS DETERMINED; ABOUT 45% OF THESE CPGS WERE LOCATED IN MORE THAN 23,000 CPG ISLANDS (CGIS). WHILE GLOBAL CPG METHYLATION WAS SIMILAR BETWEEN CLL AND NORMAL B-CELLS, 1764 GENE PROMOTERS WERE IDENTIFIED AS BEING DIFFERENTIALLY METHYLATED IN AT LEAST ONE CLL SAMPLE WHEN COMPARED WITH NORMAL B-CELL SAMPLES. NINETEEN PERCENT OF THE DIFFERENTIALLY METHYLATED GENES WERE INVOLVED IN TRANSCRIPTIONAL REGULATION. ABERRANT HYPERMETHYLATION WAS FOUND IN ALL HOX GENE CLUSTERS AND A SIGNIFICANT NUMBER OF WNT SIGNALING PATHWAY GENES. HYPOMETHYLATION OCCURRED MORE FREQUENTLY IN THE GENE BODY INCLUDING INTRONS, EXONS, AND 3'-UTRS IN CLL. THE NFATC1 P2 PROMOTER AND FIRST INTRON WAS FOUND TO BE HYPOMETHYLATED AND CORRELATED WITH UPREGULATION OF BOTH NFATC1 RNA AND PROTEIN EXPRESSION LEVELS IN CLL SUGGESTING THAT AN EPIGENETIC MECHANISM IS INVOLVED IN THE CONSTITUTIVE ACTIVATION OF NFAT ACTIVITY IN CLL CELLS. THIS COMPREHENSIVE DNA METHYLATION ANALYSIS WILL FURTHER OUR UNDERSTANDING OF THE EPIGENETIC CONTRIBUTION TO CELLULAR DYSFUNCTION IN CLL.	2012