1 5384 136 REDIFFERENTIATION OF RADIOIODINE-REFRACTORY THYROID CANCERS. THE MANAGEMENT OF RADIOIODINE REFRACTORY THYROID CANCERS (RAIR TC) IS CHALLENGING FOR THE CLINICIAN. TYROSINE KINASE INHIBITORS CLASSICALLY PRESCRIBED IN THIS SETTING CAN FAIL DUE TO PRIMARY OR ACQUIRED RESISTANCE OR THE NECESSITY OF DRUG WITHDRAWAL BECAUSE OF SERIOUS OR MODERATE BUT CHRONIC AND DELETERIOUS ADVERSE EFFECTS. THUS, THE CONCEPT OF REDIFFERENTIATION STRATEGY, WHICH INVOLVES TREATING PATIENTS WITH ONE OR MORE DRUGS CAPABLE OF RESTORING RADIOIODINE SENSITIVITY FOR RAIR TC, HAS EMERGED. THE AREA OF REDIFFERENTIATION STRATEGY LEADS TO THE CREATION OF NEW DEFINITIONS OF RAIR TC INCLUDING PERSISTENT NON RADIOIODINE-AVID PATIENTS AND 'TRUE' RAIR TC PATIENTS. THE LATTER GROUP PRESENTS A RESTORED OR INCREASED RADIOIODINE UPTAKE IN METASTATIC LESIONS BUT WITH NO RADIOLOGICAL RESPONSE ON CONVENTIONAL IMAGING, THAT IS, PROGRESSION OF A METASTATIC DISEASE, THUS PROVING THAT THEY ARE 'TRULY' RESISTANT TO THE RADIATION DELIVERED BY RADIOIODINE. UNLIKE THESE PATIENTS, METASTATIC TC PATIENTS WITH RESTORED RADIOIODINE UPTAKE OFFER THE HOPE OF PROLONGED REMISSION OR EVEN CURE OF THE DISEASE AS FOR RADIOIODINE-AVID METASTATIC TC. HERE, WE REVIEW THE DIFFERENT REDIFFERENTIATION STRATEGIES BASED ON THE UNDERLYING MOLECULAR MECHANISM LEADING TO THE SODIUM IODIDE SYMPORTER (NIS) AND RADIOIODINE UPTAKE REINDUCTION, THAT IS, BY MODULATING SIGNALING PATHWAYS, NIS TRANSCRIPTION, NIS TRAFFICKING TO THE PLASMA MEMBRANE, NIS POST-TRANSCRIPTIONAL REGULATION, BY GENE THERAPY AND OTHER POTENTIAL STRATEGIES. WE DISCUSS CLINICAL TRIALS AND PROMISING PRECLINICAL DATA OF POTENTIAL FUTURE TARGETS. 2020 2 5843 23 STUDIES ON THE CARCINOGENICITY OF POTASSIUM IODIDE IN F344 RATS. A CHRONIC TOXICITY AND CARCINOGENICITY STUDY, IN WHICH MALE AND FEMALE F344/DUCRJ RATS WERE GIVEN POTASSIUM IODIDE (KI) IN THE DRINKING WATER AT CONCENTRATIONS OF 0, 10, 100 OR 1000 PPM FOR 104 WEEKS, AND A TWO-STAGE CARCINOGENICITY STUDY OF APPLICATION AT 0 OR 1000 PPM FOR 83 WEEKS FOLLOWING A SINGLE INJECTION OF N-BIS(2-HYDROXYPROPYL)NITROSAMINE (DHPN), WERE CONDUCTED. IN THE FORMER, SQUAMOUS CELL CARCINOMAS WERE INDUCED IN THE SALIVARY GLANDS OF THE 1000 PPM GROUP, BUT NO TUMORS WERE OBSERVED IN THE THYROID. IN THE TWO-STAGE CARCINOGENICITY STUDY, THYROIDAL WEIGHTS AND THE INCIDENCE OF THYROID TUMORS DERIVED FROM THE FOLLICULAR EPITHELIUM WERE SIGNIFICANTLY INCREASED IN THE DHPN+KI AS COMPARED WITH THE DHPN ALONE GROUP. THE RESULTS OF OUR STUDIES SUGGEST THAT EXCESS KI HAS A THYROID TUMOR-PROMOTING EFFECT, BUT KI PER SE DOES NOT INDUCE THYROID TUMORS IN RATS. IN THE SALIVARY GLAND, KI WAS SUGGESTED TO HAVE CARCINOGENIC POTENTIAL VIA AN EPIGENETIC MECHANISM, ONLY ACTIVE AT A HIGH DOSE. 2000 3 479 28 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 4 5764 22 SORAFENIB ATTENUATES FIBROTIC HEPATIC INJURY THROUGH MEDIATING LYSINE CROTONYLATION. BACKGROUND: LIVER FIBROSIS IS AN INDEPENDENT CONTRIBUTOR OF CHRONIC LIVER DISEASES, AND REGRESSING LIVER FIBROSIS IS CONSIDERED A POTENTIAL THERAPEUTIC TARGET FOR CHRONIC LIVER DISEASES. WE AIMED TO EXPLORE THE EFFECTS AND MECHANISM OF SORAFENIB IN LIVER FIBROSIS. METHODS: MALE SPRAGUE DAWLEY (SD) RATS WERE SUBJECTED TO SUBCUTANEOUS INJECTION OF CARBON TETRACHLORIDE (CCL(4)) FOR 8 WEEKS TO INDUCE LIVER FIBROSIS AND THEN TREATED WITH SORAFENIB. THE DEGREE OF LIVER FIBROSIS WAS ANALYZED BY HEMATOXYLIN-EOSIN (H&E) STAINING, MASSON STAINING, AND PICROSIRIUS RED (PSR) STAINING. SERUM BIOCHEMICAL INDEXES WERE DETECTED BY FULLY AUTOMATIC BIOCHEMICAL ANALYZER OR ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA). QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QRT-PCR) WAS PERFORMED TO DETECT THE EXPRESSION OF PRO-FIBROTIC GENES. IMMUNOHISTOCHEMICAL STAINING AND WESTERN BLOTTING WERE CARRIED OUT TO EVALUATE THE LEVELS OF LYSINE CROTONYLATION. RESULTS: LIVER INDEX WAS REDUCED WITH ORAL SORAFENIB IN CCL(4)-INDUCED RATS. SERUM LIVER FUNCTION (ALANINE AMINOTRANSFERASE (ALT), ASPARTATE AMINOTRANSFERASE (AST), AND TOTAL BILIRUBIN (TBIL)) AND FIBROSIS INDICATORS (TYPE III PROCOLLAGEN (PC-III), HYALURONIC ACID (HA), AND LAMININ (LN)) WERE ATTENUATED WITH SORAFENIB TREATMENT. SORAFENIB IMPROVED THE HEPATIC STRUCTURE AND FIBROTIC PROGRESSION. THE EXPRESSION OF FIBROSIS-RELATED GENES WAS REMARKELY REDUCED WITH SORAFENIB TREATMENT. MEANWHILE, SORAFENIB INHIBITED ALPHA-SMA AND COLLAGEN I CUMULATION INDUCED BY CCL(4) INJECTION. BESIDES, PROTEIN LYSINE CROTONYLATION ESPECIALLY THE CROTONYLATED H2BK12 (H2BK12CR) AND CROTONYLATED H3K18 (H3K18CR) WERE REVERSED BY SORAFENIB, WHICH WERE DECREASED IN RESPONSE TO CCL(4) TREATMENT. SPEARMAN CORRELATION ANALYSIS SHOWN LYSINE CROTONYLATION EXPRESSION WAS NEGATIVELY CORRELATED WITH SERUM FIBROTIC INDICATORS. CONVERSELY, CROTONYLATION-REGULATED ENZYMES, WHICH NEGATIVELY REGULATE PROTEIN CROTONYLATION, WERE INCREASED IN RESPONSE TO CCL(4) TREATMENT, WHILE SORAFENIB REDUCED THEIR EXPRESSION. CONCLUSION: SORAFENIB EXERTS SIGNIFICANT ANTI-FIBROTIC EFFECTS THROUGH MEDIATING CROTONYLATION-REGULATED ENZYMES AND PROTEIN CROTONYLATION IN FIBROTIC RATS. 2022 5 4819 20 OCCURRENCE OF TOXICITY AND CELL PROLIFERATION AFTER A SINGLE GAVAGE ADMINISTRATION OF CHLOROFORM TO MALE F344 RATS. CHLOROFORM, AN INDUSTRIAL SOLVENT AND ONE OF THE MOST COMMON ENVIRONMENTAL CONTAMINANTS WHICH PRODUCES CARCINOGENIC EFFECTS IN THE LIVER AND KIDNEY OF RODENTS, IS NOT GENOTOXIC IN MOST TRADITIONAL BACTERIAL AND MAMMALIAN TEST SYSTEMS. ITS CARCINOGENIC POTENTIAL APPEARS ATTRIBUTABLE TO THE SUSTAINED CELL TURNOVER (REGENERATIVE HYPERPLASIA) WHICH RESULTS FROM CHRONIC CHLOROFORM TOXICITY. IN THIS PRESENT STUDY, CELL PROLIFERATION (REPLICATIVE DNA SYNTHESIS, RDS) AND HISTOPATHOLOGICAL CHANGES IN HEPATOCYTES AND RENAL TUBULAR EPITHELIAL CELLS WERE ASSESSED IN MALE F344 RATS FOLLOWING A SINGLE GAVAGE CHLOROFORM EXPOSURE (50, 150 OR 500 MG/KG). IN ADDITION, BIOCHEMICAL PARAMETERS (BUN, GOT, LDH AND NAG) WERE EXAMINED USING PLASMA AND URINE SAMPLES. CELL PROLIFERATION AND HISTOPATHOLOGICAL CHANGES (E.G. HYPERTROPHY, NECROSIS, VACUOLATION) WERE ONLY SEEN AT THE DOSE OF 500 MG/KG IN THE LIVER AND KIDNEY. AT THE SAME DOSE, ALL BIOCHEMICAL MARKERS WERE INCREASED AT THE 24 TO 48 HR TIME POINTS. THESE RESULTS OBTAINED ARE THUS IN LINE WITH EARLIER FINDINGS POINTING TO EPIGENETIC CARCINOGENICITY. 1998 6 1495 22 DNA HYPERMETHYLATION OF CELL CYCLE (P15 AND P16) AND APOPTOTIC (P14, P53, DAPK AND TMS1) GENES IN PERIPHERAL BLOOD OF LEUKEMIA PATIENTS. ABERRANT DNA METHYLATION OF TUMOR SUPPRESSOR GENES HAS BEEN REPORTED IN ALL MAJOR TYPES OF LEUKEMIA WITH POTENTIAL INVOLVEMENT IN THE INACTIVATION OF REGULATORY CELL CYCLE AND APOPTOSIS GENES. HOWEVER, MOST OF THE PREVIOUS REPORTS DID NOT SHOW THE EXTENT OF CONCURRENT METHYLATION OF MULTIPLE GENES IN THE FOUR LEUKEMIA TYPES. HERE, WE ANALYZED SIX KEY GENES (P14, P15, P16, P53, DAPK AND TMS1) FOR DNA METHYLATION USING METHYLATION SPECIFIC PCR TO ANALYZE PERIPHERAL BLOOD OF 78 LEUKEMIA PATIENTS (24 CML, 25 CLL, 12 AML, AND 17 ALL) AND 24 HEALTHY VOLUNTEERS. IN CML, METHYLATION WAS DETECTED FOR P15 (11%), P16 (9%), P53 (23%) AND DAPK (23%), IN CLL, P14 (25%), P15 (19%), P16 (12%), P53 (17%) AND DAPK (36%), IN AML, P14 (8%), P15 (45%), P53 (9%) AND DAPK (17%) AND IN ALL, P15 (14%), P16 (8%), AND P53 (8%). THIS STUDY HIGHLIGHTED AN ESSENTIAL ROLE OF DAPK METHYLATION IN CHRONIC LEUKEMIA IN CONTRAST TO P15 METHYLATION IN THE ACUTE CASES, WHEREAS TMS1 HYPERMETHYLATION WAS ABSENT IN ALL CASES. FURTHERMORE, HYPERMETHYLATION OF MULTIPLE GENES PER PATIENT WAS OBSERVED, WITH OBVIOUS SELECTIVENESS IN THE 9P21 CHROMOSOMAL REGION GENES (P14, P15 AND P16). INTERESTINGLY, METHYLATION OF P15 INCREASED THE RISK OF METHYLATION IN P53, AND VICE VERSA, BY FIVE FOLDS (P=0.03) INDICATING POSSIBLE SYNERGISTIC EPIGENETIC DISRUPTION OF DIFFERENT PHASES OF THE CELL CYCLE OR BETWEEN THE CELL CYCLE AND APOPTOSIS. THE INVESTIGATION OF MULTIPLE RELATIONSHIPS BETWEEN METHYLATED GENES MIGHT SHED LIGHT ON TUMOR SPECIFIC INACTIVATION OF THE CELL CYCLE AND APOPTOTIC PATHWAYS. 2014 7 4231 20 METHYLATION OF PROTOCADHERIN 10, A NOVEL TUMOR SUPPRESSOR, IS ASSOCIATED WITH POOR PROGNOSIS IN PATIENTS WITH GASTRIC CANCER. BACKGROUND & AIMS: BY USING METHYLATION-SENSITIVE REPRESENTATIONAL DIFFERENCE ANALYSIS, WE IDENTIFIED PROTOCADHERIN 10 (PCDH10), A GENE THAT ENCODES A PROTOCADHERIN AND IS SILENCED IN A TUMOR-SPECIFIC MANNER. WE ANALYZED ITS EPIGENETIC INACTIVATION, BIOLOGICAL EFFECTS, AND PROGNOSTIC SIGNIFICANCE IN GASTRIC CANCER. METHODS: METHYLATION STATUS WAS EVALUATED BY COMBINED BISULFITE RESTRICTION ANALYSIS AND BISULFITE SEQUENCING. THE EFFECTS OF PCDH10 RE-EXPRESSION WERE DETERMINED IN GROWTH, APOPTOSIS, PROLIFERATION, AND INVASION ASSAYS. PCDH10 TARGET GENES WERE IDENTIFIED BY COMPLEMENTARY DNA MICROARRAY ANALYSIS. RESULTS: PCDH10 WAS SILENCED OR DOWN-REGULATED IN 94% (16 OF 17) OF GASTRIC CANCER CELL LINES; EXPRESSION LEVELS WERE RESTORED BY EXPOSURE TO DEMETHYLATING AGENTS. RE-EXPRESSION OF PCDH10 IN MKN45 GASTRIC CANCER CELLS REDUCED COLONY FORMATION IN VITRO AND TUMOR GROWTH IN MICE; IT ALSO INHIBITED CELL PROLIFERATION (P < .01), INDUCED CELL APOPTOSIS (P < .001), AND REPRESSED CELL INVASION (P < .05), UP-REGULATING THE PRO-APOPTOSIS GENES FAS, CASPASE 8, JUN, AND CDKN1A; THE ANTIPROLIFERATION GENE FGFR; AND THE ANTI-INVASION GENE HTATIP2. PCDH10 METHYLATION WAS DETECTED IN 82% (85 OF 104) OF GASTRIC TUMORS COMPARED WITH 37% (38 OF 104) OF PAIRED NONTUMOR TISSUES (P < .0001). IN THE LATTER, PCDH10 METHYLATION WAS HIGHER IN PRECANCEROUS LESIONS (27 OF 45; 60%) THAN IN CHRONIC GASTRITIS SAMPLES (11 OF 59; 19%) (P < .0001). AFTER A MEDIAN FOLLOW-UP PERIOD OF 16.8 MONTHS, MULTIVARIATE ANALYSIS REVEALED THAT PATIENTS WITH PCDH10 METHYLATION IN ADJACENT NONTUMOR AREAS HAD A SIGNIFICANT DECREASE IN OVERALL SURVIVAL. KAPLAN-MEIER SURVIVAL CURVES SHOWED THAT PCDH10 METHYLATION WAS ASSOCIATED SIGNIFICANTLY WITH SHORTENED SURVIVAL IN STAGE I-III GASTRIC CANCER PATIENTS. CONCLUSIONS: PCDH10 IS A GASTRIC TUMOR SUPPRESSOR; ITS METHYLATION AT EARLY STAGES OF GASTRIC CARCINOGENESIS IS AN INDEPENDENT PROGNOSTIC FACTOR. 2009 8 6415 23 THE STUDY OF P16 AND P15 GENE METHYLATION IN HEAD AND NECK SQUAMOUS CELL CARCINOMA AND THEIR QUANTITATIVE EVALUATION IN PLASMA BY REAL-TIME PCR. EPIGENETIC SILENCING OF THE P16 AND P15 GENES BY PROMOTER METHYLATION ARE COMMONLY OBSERVED IN HUMAN EPITHELIAL MALIGNANCIES, INCLUDING HEAD AND NECK SQUAMOUS CELL CARCINOMAS (HNSCC). IN THIS STUDY, A METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) WAS USED TO EVALUATE THE METHYLATION STATUS OF THE P16 AND P15 GENES IN 73 HNSCC SURGICAL SPECIMENS. P16 AND P15 GENE METHYLATION WAS ALSO EXAMINED IN 29 PAIRED METASTATIC LYMPH NODES AND 29 PAIRED HISTOLOGICALLY, NORMAL RESECTION MARGIN MUCOSAE. THE QUANTITY OF CELL-FREE METHYLATED P16 AND P15 DNA IN THE PLASMA SAMPLES OF 20 HNSCC PATIENTS AND 24 HEALTHY CONTROLS WAS ALSO EXAMINED USING A FLUORESCENCE-BASED REAL-TIME PCR ASSAY. THE FREQUENCIES OF P16 AND P15 METHYLATION IN THE PRIMARY TUMOUR WERE 49% AND 60%, RESPECTIVELY. CONCORDANT METHYLATION OF P16 AND P15 IN TUMOUR SAMPLES AND METASTATIC LYMPH NODES WAS FOUND IN 59 AND 38% OF CASES, RESPECTIVELY. A SIGNIFICANTLY HIGHER PREVALENCE OF P15 METHYLATION WAS FOUND IN HISTOLOGICALLY-NORMAL SURGICAL MARGIN EPITHELIA OF HNSCC PATIENTS WITH CHRONIC SMOKING AND DRINKING HABITS COMPARED WITH NON-SMOKERS AND NON-DRINKERS. IN ADDITION, METHYLATED P16 AND P15 DNA LEVELS WERE SIGNIFICANTLY HIGHER IN THE PLASMA OF HNSCC PATIENTS (MEAN 56 COPIES/ML PLASMA AND 65 COPIES/ML PLASMA, RESPECTIVELY) COMPARED WITH NORMAL CONTROLS (MEAN 6 COPIES/ML PLASMA AND 16 COPIES/ML PLASMA, RESPECTIVELY). IN CONCLUSION, PROMOTER METHYLATION OF THE P16 AND P15 GENES IS INVOLVED IN THE PATHOGENESIS OF HNSCC AND MAY BE RELATED TO CHRONIC SMOKING AND DRINKING. THE DIFFERENTIAL LEVELS OF METHYLATED P16 AND P15 DNA IN PLASMA MIGHT BE POTENTIAL USEFUL MARKERS IN SCREENING HIGH-RISK POPULATIONS FOR EARLY HNSCC AND MONITORING THEIR TREATMENT RESPONSE. 2003 9 614 20 BINGE ALCOHOL-INDUCED MICROVESICULAR LIVER STEATOSIS AND INJURY ARE ASSOCIATED WITH DOWN-REGULATION OF HEPATIC HDAC 1, 7, 9, 10, 11 AND UP-REGULATION OF HDAC 3. BACKGROUND: BINGE, AS WELL AS CHRONIC, ALCOHOL CONSUMPTION AFFECTS GLOBAL HISTONE ACETYLATION LEADING TO CHANGES IN GENE EXPRESSION. IT IS BECOMING INCREASINGLY EVIDENT THAT THESE HISTONE-ASSOCIATED EPIGENETIC MODIFICATIONS PLAY AN IMPORTANT ROLE IN THE DEVELOPMENT OF ALCOHOL-MEDIATED HEPATIC INJURY. METHODS: C57BL/6 MICE WERE GAVAGED 3 TIMES (12-HOUR INTERVALS) WITH ETHANOL (ETOH; 4.5 G/KG). HEPATIC HISTONE DEACETYLASE (HDAC) MRNAS WERE ASSESSED BY QRT-PCR. TOTAL HDAC ACTIVITY WAS ESTIMATED BY A COLORIMETRIC HDAC ACTIVITY/INHIBITION ASSAY. HISTONE ACETYLATION LEVELS WERE EVALUATED BY WESTERN BLOT. LIVER STEATOSIS AND INJURY WERE EVALUATED BY HISTOPATHOLOGY, PLASMA AMINOTRANSFERASE (ALT) ACTIVITY, AND LIVER TRIGLYCERIDE ACCUMULATION. EXPRESSION OF FATTY ACID SYNTHASE (FAS) AND CARNITINE PALMITOYL TRANSFERASE 1A (CPT1A) WAS ALSO EXAMINED. HDAC 9 ASSOCIATION WITH FAS PROMOTER WAS ANALYZED. RESULTS: BINGE ALCOHOL EXPOSURE RESULTED IN ALTERATIONS OF HEPATIC HDAC MRNA LEVELS. DOWN-REGULATION OF HDAC CLASS I (HDAC 1), CLASS II (HDAC 7, 9, 10), AND CLASS IV (HDAC 11) AND UP-REGULATION OF HDAC CLASS I (HDAC 3) GENE EXPRESSION WERE OBSERVED. CORRESPONDENT TO THE DECREASE IN HDAC ACTIVITY, AN INCREASE IN HEPATIC HISTONE ACETYLATION WAS OBSERVED. THESE MOLECULAR EVENTS WERE ASSOCIATED WITH MICROVESICULAR HEPATIC STEATOSIS AND INJURY CHARACTERIZED BY INCREASED HEPATIC TRIGLYCERIDES (48.02 +/- 3.83 VS. 19.90 +/- 3.48 MG/G LIVER, P < 0.05) AND ELEVATED PLASMA ALT ACTIVITY (51.98 +/- 6.91 VS. 20.8 +/- 0.62 U/L, P < 0.05). HEPATIC STEATOSIS WAS ASSOCIATED WITH AN INCREASE IN FAS AND A DECREASE IN CPT1A MRNA AND PROTEIN EXPRESSION. FAS PROMOTER ANALYSIS REVEALED THAT BINGE ETOH TREATMENT DECREASED HDAC 9 OCCUPANCY AT THE FAS PROMOTER RESULTING IN ITS TRANSCRIPTIONAL ACTIVATION. CONCLUSIONS: DEREGULATION OF HEPATIC HDAC EXPRESSION LIKELY PLAYS A MAJOR ROLE IN THE BINGE ALCOHOL-INDUCED HEPATIC STEATOSIS AND LIVER INJURY BY AFFECTING LIPOGENESIS AND FATTY ACID BETA-OXIDATION. 2012 10 2847 16 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 11 672 21 BRAF, KRAS AND HELICOBACTER PYLORI EPIGENETIC CHANGES-ASSOCIATED CHRONIC GASTRITIS IN EGYPTIAN PATIENTS WITH AND WITHOUT GASTRIC CANCER. WE AIMED TO STUDY MLH1 AND MGMT METHYLATION STATUS IN HELICOBACTER PYLORI-ASSOCIATED CHRONIC GASTRITIS IN EGYPTIAN PATIENTS WITH AND WITHOUT GASTRIC CANCER. 39 PATIENTS WERE INCLUDED IN OUR STUDY. THEY WERE DIVIDED INTO 2 GROUPS; PATIENTS WITHOUT (GROUP I) AND WITH GASTRIC ADENOCARCINOMA (GROUP II). PATIENTS WERE SUBJECTED TO CLINICAL EXAMINATION, ABDOMINAL ULTRASOUND AND UPPER ENDOSCOPY FOR GASTRIC BIOPSY. BIOPSIES WERE SUBJECTED TO UREASE TEST, HISTOLOGICAL EXAMINATION, AND DNA PURIFICATION. H. PYLORI, BRAF, KRAS, MLH1 AND MGMT METHYLATION WERE ASSESSED BY QUANTITATIVE PCR. DNA SEQUENCING WAS PERFORMED TO ASSESS BRAF AND KRAS GENES MUTATION. QPCR OF H. PYLORI WAS SIGNIFICANTLY HIGHER IN PATIENTS WITH ADENOCARCINOMA (GROUP II) THAN THOSE WITHOUT ADENOCARCINOMA (GROUP I); WITH A P < 0.001 AS WELL AS IN PATIENTS WITH AGE ABOVE 50 YEARS WITH A P VALUE = 0.008. BY APPLYING LOGISTIC REGRESSION ANALYSIS IT WAS REPORTED THAT THE H. PYLORI QPCR IS A SIGNIFICANT PREDICTOR TO THE ADENOCARCINOMA WITH OR = 1.025 (95 % CI: 1. 002-1.048), WITH SENSITIVITY OF 90 % AND SPECIFICITY OF 100 %. ADENOCARCINOMA PATIENTS HAD A SIGNIFICANTLY HIGHER MEAN AGE AND LEVELS OF H. PYLORI, BRAF, K-RAS, METHYLATED MGMT AND METHYLATED MLH1 THAN THOSE OF GASTRITIS PATIENTS. DNA SEQUENCE ANALYSIS OF BRAF (CODON 12) AND KRAS (CODON 600) HAD GENES MUTATION IN GASTRIC ADENOCARCINOMA VERSUS CHRONIC GASTRITIS. CONCLUSION: H. PYLORI MAY CAUSE EPIGENETIC CHANGES PREDISPOSING THE PATIENTS TO CANCER STOMACH. ESTIMATION OF H. PYLORI BY QPCR CAN BE A GOOD PREDICTOR TO ADENOCARCINOMA. BRAF AND KRAS GENES MUTATION WERE REVELED IN GASTRITIS AND ADENOCARCINOMA PATIENTS. 2016 12 3841 19 IRON SUPPLEMENTATION REVERSES THE REDUCTION OF HYDROXYMETHYLCYTOSINE IN HEPATIC DNA ASSOCIATED WITH CHRONIC ALCOHOL CONSUMPTION IN RATS. BACKGROUND: ALCOHOL IS KNOWN TO AFFECT TWO EPIGENETIC PHENOMENA, DNA METHYLATION AND DNA HYDROXYMETHYLATION, AND IRON IS A COFACTOR OF TEN-ELEVEN TRANSLOCATION (TET) ENZYMES THAT CATALYZE THE CONVERSION FROM METHYLCYTOSINE TO HYDROXYMETHYLCYTOSINE. IN THE PRESENT STUDY WE AIMED TO DETERMINE THE EFFECTS OF ALCOHOL ON DNA HYDROXYMETHYLATION AND FURTHER EFFECTS OF IRON ON ALCOHOL ASSOCIATED EPIGENETIC CHANGES. METHODS: TWENTY-FOUR MALE SPRAGUE-DAWLEY RATS WERE FED EITHER LIEBER-DECARLI ALCOHOL DIET (36% CALORIES FROM ETHANOL) OR LIEBER-DECARLI CONTROL DIET ALONG WITH OR WITHOUT IRON SUPPLEMENTATION (0.6% CARBONYL IRON) FOR 8 WEEKS. HEPATIC NON-HEME IRON CONCENTRATIONS WERE MEASURED BY COLORIMETRIC ASSAYS. PROTEIN LEVELS OF HEPATIC FERRITIN AND TRANSFERRIN RECEPTOR WERE DETERMINED BY WESTERN BLOTTING. METHYLCYTOSINE, HYDROXYMETHYLCYTOSINE AND UNMODIFIED CYTOSINE IN DNA WERE SIMULTANEOUSLY MEASURED BY LIQUID CHROMATOGRAPHY/MASS SPECTROMETRY METHOD. RESULTS: IRON SUPPLEMENTATION SIGNIFICANTLY INCREASED HEPATIC NON-HEME IRON CONTENTS (P < 0.05) BUT ALCOHOL ALONE DID NOT. HOWEVER, BOTH ALCOHOL AND IRON SIGNIFICANTLY INCREASED HEPATIC FERRITIN LEVELS AND DECREASED HEPATIC TRANSFERRIN RECEPTOR LEVELS (P < 0.05). ALCOHOL REDUCED HEPATIC DNA HYDROXYMETHYLATION (0.21% +/- 0.04% VS. 0.33% +/- 0.04%, P = 0.01) COMPARED TO CONTROL, WHILE IRON SUPPLEMENTATION TO ALCOHOL DIET DID NOT CHANGE DNA HYDROXYMETHYLATION. THERE WAS NO SIGNIFICANT DIFFERENCE IN METHYLCYTOSINE LEVELS, WHILE UNMODIFIED CYTOSINE LEVELS WERE SIGNIFICANTLY INCREASED IN ALCOHOL-FED GROUPS COMPARED TO CONTROL (95.61% +/- 0.08% VS. 95.26% +/- 0.12%, P = 0.03), SUGGESTING THAT ALCOHOL FURTHER INCREASES THE CONVERSION FROM HYDROXYMETHYLCYTOSINE TO UNMODIFIED CYTOSINE. CONCLUSIONS: CHRONIC ALCOHOL CONSUMPTION ALTERS GLOBAL DNA HYDROXYMETHYLATION IN THE LIVER BUT IRON SUPPLEMENTATION REVERSES THE EPIGENETIC EFFECT OF ALCOHOL. 2016 13 4903 15 P16 PROMOTER HYPERMETHYLATION IN HUMAN HEPATOCELLULAR CARCINOMA WITH OR WITHOUT HEPATITIS VIRUS INFECTION. BACKGROUND: EPIGENETIC ALTERATION THROUGH METHYLATION IS ONE OF THE MOST IMPORTANT STEPS IN CARCINOGENESIS. HOWEVER, THE RELATION BETWEEN HEPATITIS VIRUS INFECTION AND EPIGENETIC ALTERATIONS IS POORLY UNDERSTOOD. METHODS: SIXTEEN PATIENTS WITHOUT HEPATITIS B VIRUS (HBV) AND HEPATITIS C VIRUS (HCV) AND 35 PATIENTS WITH HBV OR HCV WHO UNDERWENT LIVER RESECTION FOR HEPATOCELLULAR CARCINOMA (HCC) WERE STUDIED. MUTATION OF P53 WAS DETECTED BY DIRECT SEQUENCING. METHYLATION STATUS OF P16 WAS EVALUATED IN TUMOR AND NONCANCEROUS LIVER TISSUES BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. RESULTS: IN HCC WITHOUT HBV AND HCV, P53 MUTATIONS WERE DETECTED IN 5 (31%) OF 16 HCCS. METHYLATION OF P16 PROMOTER WAS DETECTED IN 2 (25%) OF 8 MODERATELY DIFFERENTIATED HCCS, 6 (75%) OF 8 POORLY DIFFERENTIATED HCCS, AND NONE OF 16 NONCANCEROUS TISSUE SPECIMENS. IN HCC WITH HBV OR HCV, P53 MUTATIONS WERE DETECTED IN 8 (23%) OF 35 HCCS. METHYLATION OF P16 PROMOTER WAS DETECTED IN 2 (100%) OF 2 WELL-DIFFERENTIATED HCCS, 13 (76%) OF 17 MODERATELY DIFFERENTIATED HCCS, 12 (75%) OF 16 POORLY DIFFERENTIATED HCCS, AND 9 (26%) OF 35 NONCANCEROUS LIVER TISSUE SPECIMENS. CONCLUSIONS: OUR RESULTS SUGGEST THAT HEPATITIS VIRUSES MIGHT INDUCE METHYLATION OF P16 PROMOTER IN LIVER WITH CHRONIC INFLAMMATION, BEFORE APPEARANCE OF HCC. 2004 14 5435 24 RELATIVE ROLE OF METHYLATOR AND TUMOR SUPPRESSOR PATHWAYS IN ULCERATIVE COLITIS-ASSOCIATED COLON CANCER. BACKGROUND: CHRONIC ULCERATIVE COLITIS (UC) IS ASSOCIATED WITH AN INCREASED COLORECTAL CANCER RISK WHICH MAY BE SECONDARY TO REPETITIVE MUCOSAL INJURY. BOTH EPIGENETIC METHYLATION AND THE CLASSIC ADENOMA-TO-CARCINOMA SEQUENCE HAVE BEEN IMPLICATED IN THIS MALIGNANT TRANSFORMATION, BUT THE UNDERLYING MOLECULAR MECHANISMS REMAIN POORLY DEFINED. THIS STUDY COMPARES THE MOLECULAR CHARACTERISTICS OF COLITIS-ASSOCIATED AND COMMON COLORECTAL CANCERS. METHODS: NINETEEN PATIENTS WITH COLORECTAL ADENOCARCINOMAS ARISING WITHIN UC WERE MATCHED FOR AGE AND CANCER SITE WITH 54 PATIENTS WITH SPORADIC ADENOCARCINOMAS. TUMOR TISSUE WAS EXAMINED FOR BRAF MUTATIONS, CPG ISLAND METHYLATOR PHENOTYPE (CIMP), AND MLH1 PROMOTER METHYLATION. MUTATIONS OF KRAS AND P53 WERE ASSESSED BY SEQUENCING. RESULTS: PATIENT DEMOGRAPHICS WERE SIMILAR FOR THE TWO GROUPS. CIMP WAS OBSERVED IN 22% OF SPORADIC COLORECTAL CANCERS AND IN 5% OF UC CANCERS (P = 0.162). RATES OF BRAF MUTATION (4% VS 5%, P = 1.0), MLH1 METHYLATION (9% VERSUS 5%, P = 0.682), AND KRAS MUTATIONS (24% VERSUS 32%, P = 0.552) WERE SIMILAR BETWEEN THE GROUPS. HOWEVER, COLITIS-ASSOCIATED COLORECTAL CANCERS WERE MORE LIKELY TO HAVE A P53 MUTATION COMPARED TO SPORADIC ADENOCARCINOMAS (95% VERSUS 53%, P = 0.001). THE DOMINANT MUTATION FOR COLITIS-ASSOCIATED CANCERS WAS A MUTATION IN CODON 4, REPRESENTING HALF OF THE MUTATIONS. FURTHERMORE, COLITIS-ASSOCIATED CANCERS HAD A HIGHER RATE OF MUTATION IN CODON 8 (48% VERSUS 6%, P < 0.001) THAN SPORADIC COUNTERPARTS. CONCLUSIONS: UNLIKE OTHER INFLAMMATORY GASTROINTESTINAL CANCERS, COLITIS-ASSOCIATED COLORECTAL CANCERS DO NOT PREFERENTIALLY ARISE VIA A METHYLATOR PATHWAY WHEN COMPARED TO SPORADIC COLORECTAL CANCERS. CHROMOSOMAL INSTABILITY REMAINS AN IMPORTANT ETIOLOGY, BUT WITH A UNIQUE P53 FREQUENCY AND MUTATION PATTERN. 2011 15 5480 31 RESVERATROL REVERSES MORPHINE-INDUCED NEUROINFLAMMATION IN MORPHINE-TOLERANT RATS BY REVERSAL HDAC1 EXPRESSION. BACKGROUND/PURPOSE: WE PREVIOUSLY SHOWED THAT SUBSEQUENT INTRATHECAL (I.T.) INJECTION OF RESVERATROL (30 MUG) SIGNIFICANTLY REVERSES MORPHINE-EVOKED NEUROINFLAMMATION IN MORPHINE-TOLERANT RATS. THE PRESENT STUDY EXAMINED THE UNDERLYING MECHANISM. METHODS: MALE WISTAR RATS WERE IMPLANTED WITH TWO I.T. CATHETERS, ONE OF WHICH WAS CONNECTED TO A MINIOSMOTIC PUMP AND USED FOR MORPHINE (15 MUG/H) OR SALINE INFUSION FOR 120 HOURS. TO EXAMINE THE EFFECTS ON SPINAL CORD EXPRESSION OF HISTONE DEACETYLASE 1 (HDAC1), THE INFLAMMATORY CYTOKINE TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA), AND TNF RECEPTOR (TNFR) 1 AND TNFR2 DURING TOLERANCE INDUCTION, A TAIL-FLICK TEST WAS PERFORMED PRIOR TO INFUSION AND AFTER 24 HOURS, 48 HOURS, 72 HOURS, 96 HOURS, AND 120 HOURS OF INFUSION. RESULTS: RESVERATROL TREATMENT PRIOR TO MORPHINE CHALLENGE RESTORED THE ANTINOCICEPTIVE EFFECT OF MORPHINE IN MORPHINE-TOLERANT RATS AND REVERSED THE MORPHINE INFUSION-INDUCED INCREASE IN HDAC1, TNF-ALPHA, AND TNFR1 EXPRESSION. MOREOVER, CHRONIC MORPHINE INFUSION INCREASED TNFR1-SPECIFIC EXPRESSION IN NEURON IN MORPHINE-TOLERANT RAT SPINAL CORDS, AND THIS EFFECT WAS ALMOST COMPLETELY INHIBITED BY RESVERATROL TREATMENT PRIOR TO MORPHINE CHALLENGE. CONCLUSION: RESVERATROL RESTORES THE ANTINOCICEPTIVE EFFECT OF MORPHINE BY REVERSING MORPHINE INFUSION-INDUCED SPINAL CORD NEUROINFLAMMATION AND INCREASE IN TNFR1 EXPRESSION. THE REVERSAL OF THE MORPHINE-INDUCED INCREASE IN TNFR1 EXPRESSION BY RESVERATROL IS PARTIALLY DUE TO REVERSAL OF THE MORPHINE INFUSION-INDUCED INCREASE IN HDAC1 EXPRESSION. RESVERATROL PRETREATMENT CAN BE USED AS AN ADJUVANT IN CLINICAL PAIN MANAGEMENT FOR PATIENTS WHO NEED LONG-TERM MORPHINE TREATMENT OR WITH NEUROPATHIC PAIN. 2016 16 1968 26 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 17 2135 34 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 18 1213 24 CPG ISLAND METHYLATION PATTERNS IN CHRONIC LYMPHOCYTIC LEUKEMIA. CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS THE MOST COMMON LEUKEMIA IN WESTERN COUNTRIES. IN CLL, A LARGE NUMBER OF GENES AFFECTING CANCER-RELATED PATHWAYS MAY BE DYSREGULATED BY EPIGENETIC SILENCING. WE ANALYSED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION THE CPG ISLAND METHYLATION STATUS OF 15 WELL-CHARACTERISED CANCER-RELATED GENES IN 32 PATIENTS WITH CLL. ABERRANT METHYLATION IN THE SAMPLE OF PATIENTS WITH CLL WAS SHOWN FOR SECRETED FRIZZLED-RELATED PROTEIN 1 (68.8%), SECRETED FRIZZLED-RELATED PROTEIN 2 (65.6%), DEATH-ASSOCIATED PROTEIN KINASE 1 (50.0%), E-CADHERIN (21.9%), SECRETED FRIZZLED-RELATED PROTEIN 4 (15.6%), P15 (9.4%), P16 (6.3%), RETINOIC ACID RECEPTOR BETA2 (3.1%), SECRETED FRIZZLED-RELATED PROTEIN 5 (3.1%) AND TISSUE INHIBITOR OF MATRIX METALLOPROTEINASES 3 (3.1%). FOR HUMAN MUT-L HOMOLOG 1, O(6)-METHYLGUANINE DNA METHYLTRANSFERASE, P73, SUPPRESSOR OF CYTOKINE SIGNALLING 1 AND TISSUE INHIBITOR OF MATRIX METALLOPROTEINASES 2 NO HYPERMETHYLATION WAS DETECTED. HYPERMETHYLATION OF AT LEAST ONE GENE WAS OBSERVED IN 87.5% OF THE SAMPLES. OUR RESULTS SHOW THAT ABERRANT CPG ISLAND METHYLATION AFFECTING CANCER-RELATED PATHWAYS SUCH AS WNT SIGNALLING, REGULATION OF APOPTOSIS, CELL CYCLE CONTROL AND TISSUE INVASION IS A COMMON PHENOMENON IN CLL. EPIGENETIC DISTURBANCES MAY BE INVOLVED IN THE PATHOGENESIS OF CLL AND THUS MAY PROVIDE A MOLECULAR RATIONALE FOR THERAPEUTIC APPROACHES. 2009 19 779 20 CELL-FREE DNA PROMOTER HYPERMETHYLATION AS A DIAGNOSTIC MARKER FOR PANCREATIC DUCTAL ADENOCARCINOMA - AN EXTERNAL VALIDATION STUDY. BACKGROUND: WE RECENTLY IDENTIFIED A DIAGNOSTIC PREDICTION MODEL BASED ON PROMOTER HYPERMETHYLATION OF EIGHT SELECTED GENES IN PLASMA CELL-FREE (CF) DNA, WHICH SHOWED PROMISING RESULTS AS A DIAGNOSTIC BIOMARKER FOR PANCREATIC DUCTAL ADENOCARCINOMA (PDAC). THE AIM OF THE PRESENT STUDY WAS TO VALIDATE THIS BIOMARKER PROFILE IN AN EXTERNAL PATIENT COHORT AND EXAMINE ANY ADDITIONAL EFFECT OF SERUM CA 19-9. METHODS: PATIENTS WITH PDAC (N = 346, STAGE I-IV) AND CHRONIC PANCREATITIS (N = 25) WERE INCLUDED. METHYLATION-SPECIFIC PCR OF A 28-GENE PANEL WAS PERFORMED ON SERUM CFDNA SAMPLES. THE PREVIOUSLY DEVELOPED DIAGNOSTIC PREDICTION MODEL (AGE>65 YEARS, BMP3, RASSF1A, BNC1, MESTV2, TFPI2, APC, SFRP1 AND SFRP2) WAS VALIDATED ALONE AND IN COMBINATION WITH SERUM CA 19-9 IN THIS EXTERNAL PATIENT COHORT. RESULTS: PATIENTS WITH PDAC HAD A HIGHER NUMBER OF HYPERMETHYLATED GENES (MEAN 8.11, 95% CI 7.70-8.52) THAN PATIENTS WITH CHRONIC PANCREATITIS (MEAN 5.60, 95% CI 4.42-6.78, P = 0.011). VALIDATION OF THE DIAGNOSTIC PREDICTION MODEL YIELDED AN AUC OF 0.77 (95% CI 0.69-0.84). THE COMBINATION OF SERUM CA 19-9 AND OUR TEST HAD AN AUC OF 0.93 (95% CI 0.89-0.96) IN THE PRIMARY STUDY AND 0.85 (95% CI 0.79-0.91) IN THE VALIDATION STUDY. CONCLUSION: IN THIS VALIDATION STUDY, PDAC WAS ASSOCIATED WITH A HIGHER NUMBER OF HYPERMETHYLATED GENES IN SERUM CFDNA THAN CHRONIC PANCREATITIS. OUR DIAGNOSTIC TEST WAS SUPERIOR TO THE PREDICTIVE VALUE OF SERUM CA 19-9 ALONE IN BOTH THE PRIMARY AND THE VALIDATION STUDY. THE COMBINATION OF OUR TEST WITH CA 19-9 MAY SERVE AS A CLINICALLY USEFUL DIAGNOSTIC BIOMARKER FOR PDAC. 2021 20 401 24 ANALYSIS OF ABERRANT METHYLATION ON PROMOTER SEQUENCES OF TUMOR SUPPRESSOR GENES AND TOTAL DNA IN SPUTUM SAMPLES: A PROMISING TOOL FOR EARLY DETECTION OF COPD AND LUNG CANCER IN SMOKERS. BACKGROUND: CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS A DISORDER ASSOCIATED TO CIGARETTE SMOKE AND LUNG CANCER (LC). SINCE EPIGENETIC CHANGES IN ONCOGENES AND TUMOR SUPPRESSOR GENES (TSGS) ARE CLEARLY IMPORTANT IN THE DEVELOPMENT OF LC. IN THIS STUDY, WE HYPOTHESIZE THAT TOBACCO SMOKERS ARE SUSCEPTIBLE FOR METHYLATION IN THE PROMOTER REGION OF TSGS IN AIRWAY EPITHELIAL CELLS WHEN COMPARED WITH NON-SMOKER SUBJECTS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE USEFULNESS OF DETECTION OF GENES PROMOTER METHYLATION IN SPUTUM SPECIMENS, AS A COMPLEMENTARY TOOL TO IDENTIFY LC BIOMARKERS AMONG SMOKERS WITH EARLY COPD. METHODS: WE DETERMINED THE AMOUNT OF DNA IN INDUCED SPUTUM FROM PATIENTS WITH COPD (N = 23), LC (N = 26), AS WELL AS IN HEALTHY SUBJECTS (CTR) (N = 33), USING A COMMERCIAL KIT FOR DNA PURIFICATION, FOLLOWED BY ABSORBANCE MEASUREMENT AT 260 NM. THE FREQUENCY OF CDKN2A, CDH1 AND MGMT PROMOTER METHYLATION IN THE SAME GROUPS WAS DETERMINED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP). THE FISHER'S EXACT TEST WAS EMPLOYED TO COMPARE FREQUENCY OF RESULTS BETWEEN DIFFERENT GROUPS. RESULTS: DNA CONCENTRATION WAS 7.4 AND 5.8 TIMES HIGHER IN LC AND COPD COMPARED TO THE (CTR) (P < 0.0001), RESPECTIVELY. METHYLATION STATUS OF CDKN2A AND MGMT WAS SIGNIFICANTLY HIGHER IN COPD AND LC PATIENTS COMPARED WITH CTR GROUP (P < 0.0001). FREQUENCY OF CDH1 METHYLATION ONLY SHOWED A STATISTICALLY SIGNIFICANT DIFFERENCE BETWEEN LC PATIENTS AND CTR GROUP (P < 0.05). CONCLUSIONS: WE PROVIDE EVIDENCE THAT ABERRANT METHYLATION OF TSGS IN SAMPLES OF INDUCED SPUTUM IS A USEFUL TOOL FOR EARLY DIAGNOSTIC OF LUNG DISEASES (LC AND COPD) IN SMOKER SUBJECTS. VIRTUAL SLIDES: THE ABSTRACT MUST FINISH WITH THE FOLLOWING TEXT: VIRTUAL SLIDES THE VIRTUAL SLIDE(S) FOR THIS ARTICLE CAN BE FOUND HERE: HTTP://WWW.DIAGNOSTICPATHOLOGY.DIAGNOMX.EU/VS/1127865005664160. 2012