1 4245 123 METHYLATION STATUS OF DDIT3 GENE IN CHRONIC MYELOID LEUKEMIA. BACKGROUND: DNA-DAMAGE-INDUCIBLE TRANSCRIPT 3 (DDIT3), A CANDIDATE TUMOR SUPPRESSOR GENE (TSG), HAS BEEN FOUND INVOLVED IN THE REGULATION OF CELLULAR GROWTH AND DIFFERENTIATION. THE EPIGENETIC CHANGES OF TSGS ARE RECENTLY RECOGNIZED AS AN ABNORMAL MECHANISM CONTRIBUTING TO THE DEVELOPMENT OF CHRONIC MYELOID LEUKEMIA (CML). THE AIM OF THIS STUDY WAS TO INVESTIGATE THE METHYLATION STATUS OF DDIT3 GENE IN CML PATIENTS. METHODS: THE METHYLATION STATUS OF DDIT3 PROMOTER WAS DETECTED IN THE BONE MARROW MONONUCLEAR CELLS FROM 53 PATIENTS WITH CML USING METHYLATION-SPECIFIC PCR (MSP). THE EXPRESSION LEVELS OF DDIT3 AND BCR/ABL TRANSCRIPT WERE DETERMINED BY REAL-TIME QUANTITATIVE PCR (RQ-PCR). CLINICAL DATA OF THESE PATIENTS WERE COLLECTED AND ANALYZED. RESULTS: THE ABERRANT METHYLATION OF DDIT3 GENE PROMOTER WAS FOUND IN 35 OF 53 (66%) CML CASES. CORRELATION WAS NOT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND THE AGE, SEX, HEMOGLOBIN CONCENTRATION, PLATELET COUNTS, CHROMOSOMAL ABNORMALITIES, BCR/ABL TRANSCRIPT, AND STAGING OF CML PATIENTS (P > 0.05), BUT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND WBC COUNTS OF CML CASES (R = 0.781, P < 0.001). THE LEVEL OF DDIT3 TRANSCRIPT IN CML PATIENTS WAS SIGNIFICANTLY LOWER THAN THAT IN CONTROLS (MEDIAN 3.28 VS 19.69, P < 0.001), HOWEVER, THERE WAS NO DIFFERENCE IN THE LEVEL OF DDIT3 TRANSCRIPT BETWEEN METHYLATION-POSITIVE CML CASES (0.05-65.32, MEDIAN 2.13) AND METHYLATION- NEGATIVE CML CASES (0.12-126.04, MEDIAN 3.92) (P > 0.05). CONCLUSION: OUR RESULTS DEMONSTRATE THAT ABERRANT METHYLATION OF DDIT3 OCCURS IN CML FREQUENTLY. 2010 2 156 61 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 3 3083 34 GENOME-WIDE SCREEN OF DNA METHYLATION CHANGES INDUCED BY LOW DOSE X-RAY RADIATION IN MICE. EPIGENETIC MECHANISMS PLAY A KEY ROLE IN NON-TARGETED EFFECTS OF RADIATION. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE GLOBAL HYPOMETHYLATION AND PROMOTER HYPERMETHYLATION OF PARTICULAR GENES INDUCED BY LOW DOSE RADIATION (LDR). THIRTY MALE BALB/C MICE WERE DIVIDED INTO 3 GROUPS: CONTROL, ACUTELY EXPOSED (0.5 GY X-RAYS), AND CHRONIC EXPOSURE FOR 10 DAYS (0.05GY/DX10D). HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) AND MEDIP-QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR) WERE USED TO STUDY METHYLATION PROFILES. DNMT1 AND MBD2 EXPRESSION WAS DETERMINED BY QPCR AND WESTERN BLOT ASSAYS. METHYLATION AND EXPRESSION OF RAD23B AND DDIT3 WERE DETERMINED BY BISULFATE SEQUENCING PRIMERS (BSP) AND QPCR, RESPECTIVELY. THE RESULTS SHOW THAT LDR INDUCED GENOMIC HYPOMETHYLATION IN BLOOD 2 H POSTIRRADITAION, BUT WAS NOT RETAINED AT 1-MONTH. DNMT1 AND MBD2 WERE DOWNREGULATED IN A TISSUE-SPECIFIC MANNER BUT DID NOT PERSIST. SPECIFIC HYPERMETHYLATION WAS OBSERVED FOR 811 REGIONS IN THE GROUP RECEIVING CHRONIC EXPOSURE, WHICH COVERED ALMOST ALL KEY BIOLOGICAL PROCESSES AS INDICATED BY GO AND KEGG PATHWAY ANALYSIS. EIGHT HYPERMETHYLATED GENES (RAD23B, TDG, CCND1, DDIT3, LLGL1, RASL11A, TBX2, SCL6A15) WERE VERIFIED BY MEDIP-QPCR. AMONG THEM, RAD23B AND DDIT3 GENE DISPLAYED TISSUE-SPECIFIC METHYLATION AND DOWNREGULATION, WHICH PERSISTED FOR 1-MONTH POSTIRRADIATION. THUS, LDR INDUCED GLOBAL HYPOMETHYLATION AND TISSUE-SPECIFIC PROMOTER HYPERMETHYLATION OF PARTICULAR GENES. PROMOTER HYPERMETHYLATION, RATHER THAN GLOBAL HYPOMETHYLATION, WAS RELATIVELY STABLE. DYSREGULATION OF METHYLATION MIGHT BE CORRELATED WITH DOWN-REGULATION OF DNMT1 AND MBD2, BUT MUCH BETTER UNDERSTANDING THE MOLECULAR MECHANISMS INVOLVED IN THIS PROCESS WILL REQUIRE FURTHER STUDY. 2014 4 3461 46 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 5 5270 45 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 6 2847 27 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 7 831 43 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 8 3409 40 HOXA4 GENE PROMOTER HYPERMETHYLATION AS AN EPIGENETIC MECHANISM MEDIATING RESISTANCE TO IMATINIB MESYLATE IN CHRONIC MYELOID LEUKEMIA PATIENTS. DEVELOPMENT OF RESISTANCE TO IMATINIB MESYLATE (IM) IN CHRONIC MYELOID LEUKEMIA (CML) PATIENTS HAS EMERGED AS A SIGNIFICANT CLINICAL PROBLEM. THE OBSERVATION THAT INCREASED EPIGENETIC SILENCING OF POTENTIAL TUMOR SUPPRESSOR GENES CORRELATES WITH DISEASE PROGRESSION IN SOME CML PATIENTS TREATED WITH IM SUGGESTS A RELATIONSHIP BETWEEN EPIGENETIC SILENCING AND RESISTANCE DEVELOPMENT. WE HYPOTHESIZE THAT PROMOTER HYPERMETHYLATION OF HOXA4 COULD BE AN EPIGENETIC MECHANISM MEDIATING IM RESISTANCE IN CML PATIENTS. THUS A STUDY WAS UNDERTAKEN TO INVESTIGATE THE PROMOTER HYPERMETHYLATION STATUS OF HOXA4 IN CML PATIENTS ON IM TREATMENT AND TO DETERMINE ITS ROLE IN MEDIATING RESISTANCE TO IM. GENOMIC DNA WAS EXTRACTED FROM PERIPHERAL BLOOD SAMPLES OF 95 CML PATIENTS (38 GOOD RESPONDERS AND 57 RESISTANT) AND 12 NORMAL CONTROLS. ALL SAMPLES WERE BISULFITE TREATED AND ANALYSED BY METHYLATION-SPECIFIC HIGH-RESOLUTION MELT ANALYSIS. COMPARED TO THE GOOD RESPONDERS, THE HOXA4 HYPERMETHYLATION LEVEL WAS SIGNIFICANTLY HIGHER (P = 0.002) IN IM-RESISTANT CML PATIENTS. ON COMPARING THE RISK, HOXA4 HYPERMETHYLATION WAS ASSOCIATED WITH A HIGHER RISK FOR IM RESISTANCE (OR 4.658; 95% CI, 1.673-12.971; P = 0.003). THUS, IT IS REASONABLE TO SUGGEST THAT PROMOTER HYPERMETHYLATION OF HOXA4 GENE COULD BE AN EPIGENETIC MECHANISM MEDIATING IM RESISTANCE IN CML PATIENTS. 2013 9 2135 45 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 10 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 11 4243 47 METHYLATION STATUS OF CEBPA GENE PROMOTER IN CHRONIC MYELOID LEUKEMIA. CCAAT/ENHANCER BINDING PROTEIN ALPHA IS ONE OF THE CRUCIAL TRANSCRIPTION FACTORS FOR MYELOID CELL DEVELOPMENT THAT HAS BEEN FOUND TO BE INVOLVED IN HEMATOPOIETIC DIFFERENTIATION AND LEUKEMIOGENESIS. RECENTLY, EPIGENETIC REGULATION OF CEBPA EXPRESSION THROUGH DNA METHYLATION HAS BEEN DEMONSTRATED IN LEUKEMIA. THE AIM OF THIS STUDY WAS TO INVESTIGATE THE METHYLATION STATUS OF CEBPA GENE IN CHRONIC MYELOID LEUKEMIA (CML) PATIENTS. THE METHYLATION STATUS OF CEBPA PROMOTER WAS STUDIED IN 100 PATIENTS WITH CML AND 98 NORMAL HEALTHY INDIVIDUALS FROM HYDERABAD, INDIA, USING METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. THE ABERRANT METHYLATION OF CEBPA GENE PROMOTER WAS FOUND IN 32 OF THE 100 CML CASES. A HIGHLY SIGNIFICANT ASSOCIATION WAS FOUND BETWEEN THE FREQUENCY OF CEBPA GENE PROMOTER HYPERMETHYLATION AND THE CML STAGES (P = 0.017), BUT ASSOCIATION WITH RESPECT TO AGE AND GENDER OF THE PATIENT WAS NOT FOUND. THE RESULTS SUGGEST THAT ABERRANT METHYLATION IN THE CPG ISLAND OF THE PROMOTER REGION OF THIS GENE MIGHT BE A COMMON EVENT IN CML, AND SYSTEMIC EXPRESSION STUDIES WILL BE NEEDED TO UNFOLD THE ROLE OF CEBPA PROMOTER METHYLATION IN THE DEVELOPMENT, PROGRESSION, AND PROGNOSIS OF CML. 2014 12 139 30 ABERRANT DNA METHYLATION IS ASSOCIATED WITH DISEASE PROGRESSION, RESISTANCE TO IMATINIB AND SHORTENED SURVIVAL IN CHRONIC MYELOGENOUS LEUKEMIA. THE EPIGENETIC IMPACT OF DNA METHYLATION IN CHRONIC MYELOGENOUS LEUKEMIA (CML) IS NOT COMPLETELY UNDERSTOOD. TO ELUCIDATE ITS ROLE WE ANALYZED 120 PATIENTS WITH CML FOR METHYLATION OF PROMOTER-ASSOCIATED CPG ISLANDS OF 10 GENES. FIVE GENES WERE IDENTIFIED BY DNA METHYLATION SCREENING IN THE K562 CELL LINE AND 3 GENES IN PATIENTS WITH MYELOPROLIFERATIVE NEOPLASMS. THE CDKN2B GENE WAS SELECTED FOR ITS FREQUENT METHYLATION IN MYELOID MALIGNANCIES AND ABL1 AS THE TARGET OF BCR-ABL TRANSLOCATION. THIRTY PATIENTS WERE IMATINIB-NAIVE (MOSTLY TREATED BY INTERFERON-ALPHA BEFORE THE IMATINIB ERA), 30 WERE IMATINIB-RESPONSIVE, 50 WERE IMATINIB-RESISTANT, AND 10 WERE IMATINIB-INTOLERANT. WE QUANTIFIED DNA METHYLATION BY BISULFITE PYROSEQUENCING. THE AVERAGE NUMBER OF METHYLATED GENES WAS 4.5 PER PATIENT IN THE CHRONIC PHASE, INCREASING SIGNIFICANTLY TO 6.2 IN THE ACCELERATED AND 6.4 IN THE BLASTIC PHASE. HIGHER NUMBERS OF METHYLATED GENES WERE ALSO OBSERVED IN PATIENTS RESISTANT OR INTOLERANT TO IMATINIB. THESE PATIENTS ALSO SHOWED ALMOST EXCLUSIVE METHYLATION OF A PUTATIVE TRANSPORTER OSCP1. ABNORMAL METHYLATION OF A SRC SUPPRESSOR GENE PDLIM4 WAS ASSOCIATED WITH SHORTENED SURVIVAL INDEPENDENTLY OF CML STAGE AND IMATINIB RESPONSIVENESS. WE CONCLUDE THAT ABERRANT DNA METHYLATION IS ASSOCIATED WITH CML PROGRESSION AND THAT DNA METHYLATION COULD BE A MARKER ASSOCIATED WITH IMATINIB RESISTANCE. FINALLY, DNA METHYLATION OF PDLIM4 MAY HELP IDENTIFY A SUBSET OF CML PATIENTS THAT WOULD BENEFIT FROM TREATMENT WITH SRC/ABL INHIBITORS. 2011 13 157 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 14 5458 38 RESEARCH ON EPIGENETIC MECHANISM OF SFRP2 IN ADVANCED CHRONIC MYELOID LEUKEMIA. SECRETED FRIZZLED-RELATED PROTEIN 2 (SFRP2) HAS BEEN REPORTED TO ACT AS A TUMOR SUPPRESSORS. THIS STUDY AIMS TO DETECT THE BIOLOGICAL ROLE OF SFRP2 IN ADVANCED CHRONIC MYELOID LEUKEMIA (CML). IN THIS STUDY WE EXAMINED BONE MARROW SAMPLES FROM 45 CML PATIENTS AND 10 HEALTHY DONORS. K562 AND KCL22 CELLS WERE CULTURED AND TREATED WITH DEMETHYLATION DRUG AND HISTONE DEACETYLASE INHIBITOR (HDACI). KCL22 AND K562 CELLS WERE TRANSFECTED WITH LENTIVIRAL VECTOR (LV)-SFRP2, LV-CONTROL. THE CELLS WERE THEN SUBJECTED TO PROLIFERATION AND APOPTOSIS ASSAYS, REAL TIME POLYMERASE CHAIN REACTION (PCR), METHYLATION-SPECIFIC PCR (MSP), WESTERN BLOTTING, CO-IMMUNOPRECIPITATION (COIP) AND CHROMATIN IMMUNOPRECIPITATION (CHIP), WE FOUND THAT SFRP2 WAS DOWN-REGULATED IN THE ACCELERATED AND BLAST PHASE OF CML, WHEREAS, THE LEVELS OF WNT1, WNT3 AND WNT5A WERE UP-REGULATED IN THE ACCELERATED AND BLAST PHASE OF CML. OVEREXPRESSION SFRP2 INHIBITED PROLIFERATION, PROMOTED APOPTOSIS AND ACTIVATED THE WNT PATHWAY. COIP-MS RESULTS SHOWED THAT SFRP2 INTERACTED WITH WNT1 AND WNT5A. CHIP-SEQ RESULT INDICATED THAT THE PROMOTER OF H3K4ME3 AND H3K27ME3 WERE ABLE TO INTERACT WITH SFRP2. IN CONCLUSION, OUR FINDINGS DEMONSTRATED THE SFRP2 ACT AS A POTENTIAL THERAPEUTIC TARGET FOR ADVANCED CML. FURTHERMORE, OUR RESULTS SUPPORT THE USE OF DEMETHYLATION DRUGS AND HDACI AS A POTENTIAL CML TREATMENT STRATEGY. 2018 15 3071 41 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 16 3588 49 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 17 817 37 CHARACTERISTIC PATTERNS OF ALTERED DNA METHYLATION PREDICT EMERGENCE OF HUMAN HEPATOCELLULAR CARCINOMA. WE AIMED TO IDENTIFY THE SPECIFIC SUBSET OF TUMOR SUPPRESSOR GENES (TSGS) THAT ARE METHYLATION-SILENCED DURING THE EARLIEST STEPS OF HEPATOCARCINOGENESIS, AND TO FURTHER EVALUATE WHETHER THESE GENES CAN SERVE AS PREDICTIVE BIOMARKERS OF HEPATOCELLULAR CARCINOMA (HCC) EMERGENCE. A TOTAL OF 482 LIVER TISSUES INCLUDING 177 PAIRS OF HCCS AND MATCHED NONTUMOR LIVERS AND 128 LIVER BIOPSIES FROM CHRONIC HEPATITIS C (CHC) PATIENTS WERE ANALYZED FOR QUANTITATIVE METHYLATION ANALYSIS IN 24 TSG PROMOTERS AND THREE MINT LOCI. THE TUMORS WERE CLASSIFIED AS EARLY, LESS-PROGRESSED, AND HIGHLY PROGRESSED HCCS USING HISTOLOGY AND RADIOLOGICAL APPROACHES. A SUBSET OF TSGS THAT HARBORED DISTINCTLY HIGH LEVELS OF METHYLATION IN EARLY HCCS WERE SELECTED. BASED ON THE METHYLATION PROFILES OF THESE GENES, KAPLAN-MEIER ANALYSES WERE PERFORMED TO DETERMINE TIME-TO-HCC OCCURRENCE IN CHC PATIENTS. SUBSEQUENTLY, MULTIVARIATE ANALYSIS WAS PERFORMED USING AGE, GENDER, FIBROSIS STAGE, AND NUMBER OF METHYLATED TSGS AS COVARIATES. AMONG TSGS ANALYZED, A SUBSET OF EIGHT TSGS (HIC1, GSTP1, SOCS1, RASSF1, CDKN2A, APC, RUNX3, AND PRDM2) DEMONSTRATED A DISTINCT CLUSTER BY HIERARCHICAL CLUSTERING AND RECEIVER OPERATING CHARACTERISTIC ANALYSES. THIS SUBSET OF TSGS SHOWED SIGNIFICANTLY HIGHER METHYLATION LEVELS IN THE EARLY HCCS (P < 0.0001). IN THE CHC PATIENTS, METHYLATION FREQUENCIES IN THESE TSGS WERE ASSOCIATED WITH SHORTER TIME-TO-HCC OCCURRENCE (P < 0.0001), AND NUMBER OF METHYLATED GENES WAS AN INDEPENDENT RISK FACTOR FOR HCC (HAZARD RATIO = 5.21, 95% CONFIDENCE INTERVAL = 2.25-11.76, P = 0.0002). CONCLUSION: EPIGENETIC INACTIVATION OF A SUBSET OF TSGS PLAYS A CRITICAL ROLE IN THE EARLIEST STEPS OF HEPATOCARCINOGENESIS. FURTHERMORE, EPIGENETIC INACTIVATION OF THESE GENES IN CHC PROVIDES A PROGNOSTIC VALUE FOR DETERMINING THE RISK FOR DEVELOPING HCC LATER IN LIFE. 2012 18 5669 25 SFRP1 PROMOTER METHYLATION IS ASSOCIATED WITH PERSISTENT PHILADELPHIA CHROMOSOME IN CHRONIC MYELOID LEUKEMIA. EPIGENETIC SILENCING OF SFRP GENES HAS BEEN SHOWN TO LEAD TO CONSTITUTIVE ACTIVATION OF THE CANONICAL WNT-SIGNALING PATHWAY. THE FIRST DESCRIPTION OF DEREGULATED WNT-SIGNALING ACTIVATION IN A HEMATOLOGICAL MALIGNANCY WAS REPORTED IN CHRONIC MYELOID LEUKEMIA (CML). TO INVESTIGATE WHETHER EPIGENETIC SILENCING OF SFRP IS RESPONSIBLE FOR THE OBSERVED WNT ACTIVATION IN CML, WE STUDIED THE METHYLATION AND MUTATIONAL STATUS OF THE SFRP1 PROMOTER IN 48 CHRONIC PHASE CML PATIENTS. OF THE 48 CML PATIENTS 41 WERE SHOWN TO BE UNMETHYLATED, 6 PATIENTS HEMI-METHYLATED AND 1 PATIENT FULLY METHYLATED AT THE SFRP1 PROMOTER. ALBEIT OBSERVED INFREQUENTLY IN CHRONIC PHASE CML, SFRP1 PROMOTER METHYLATION CORRELATED WITH PRIMARY CYTOGENETIC RESISTANCE TO IMATINIB MESYLATE. SFRP1 PROMOTER METHYLATION MAY INDICATE A GENETICALLY MORE UNSTABLE FORM OF DISEASE RESISTANT TO THERAPY AND PROVIDE A KEY BIOLOGICAL DIFFERENCE IN THERAPY RESISTANT PATIENTS, IN ADDITION TO A POSSIBLE MECHANISM FOR THE OBSERVED ACTIVATION OF CANONICAL WNT SIGNALING IN CML. 2009 19 494 35 ASSESSMENT OF PROMOTER METHYLATION IDENTIFIES PTCH AS A PUTATIVE TUMOR-SUPPRESSOR GENE IN HUMAN CLL. BACKGROUND: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS CHARACTERIZED BY A CLONAL ACCUMULATION OF NEOPLASTIC LYMPHOCYTES, INDICATING DISRUPTION OF APOPTOSIS. PATIENTS AND METHODS: DIFFERENTIAL METHYLATION HYBRIDIZATION ANALYSIS WAS PERFORMED TO IDENTIFY NOVEL TARGET GENES SILENCED BY CPG ISLAND METHYLATION IN PATIENTS WITH CLL. RESULTS: PATCHED (PTCH), A TUMOR-SUPPRESSOR GENE, WAS FOUND TO BE FREQUENTLY METHYLATED IN CLL SAMPLES COMPARED TO SAMPLES DERIVED FROM HEALTHY INDIVIDUALS. DE NOVO METHYLATION OF A CPG ISLAND REGION LOCATED UPSTREAM OF PTCH EXON 1 WAS CONFIRMED BY PYROSEQUENCING IN 17/37 (46%) OF PERIPHERAL BLOOD MONONUCLEAR CELLS OF PATIENTS WITH CLL, BUT IN NONE ISOLATED FROM SEVEN HEALTHY INDIVIDUALS. NO ASSOCIATION WAS FOUND BETWEEN PTCH HYPERMETHYLATION AND CURRENTLY USED PROGNOSTIC CLL FACTORS. CONCLUSION: OUR INVESTIGATION SUGGESTS THAT EPIGENETIC SILENCING OF PTCH IS A MECHANISM CONTRIBUTING TO CLL TUMORIGENESIS. 2016 20 2759 43 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