1  2842 138 FREQUENT CONCOMITANT EPIGENETIC SILENCING OF SOX1 AND SECRETED FRIZZLED-RELATED PROTEINS (SFRPS) IN HUMAN HEPATOCELLULAR CARCINOMA. BACKGROUND AND AIM: EXCEPT FOR GENETIC MUTATIONS, EPIGENETIC CHANGES ARE ALSO INVOLVED IN THE DEVELOPMENT OF HUMAN CANCERS. RECENTLY, WE HAVE IDENTIFIED SOX1, SRY (SEX DETERMINING REGION Y)-BOX 1, IS HYPERMETHYLATED IN CERVICAL CANCER AND OVARIAN CANCER. THEREFORE, WE INVESTIGATED WHETHER PROMOTER HYPERMETHYLATION OF SOX1 IS COMMON IN HEPATOCELLULAR CARCINOMA (HCC). METHODS: WE USED METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MS-PCR) AND BISULFITE SEQUENCING TO ANALYZE THE METHYALTION LEVEL OF THE SOX1 PROMOTER IN SEVEN HCC CELL LINES, 54 CLINICAL HCCS, 42 CIRRHOTIC LIVERS, 21 LIVERS WITH CHRONIC HEPATITIS, AND 15 CONTROL LIVERS. THEN, WE EMPLOYED QUANTITATIVE MS-PCR (QMSP) TO VALIDATE IN AN INDEPENDENT SET OF SAMPLES (60 PAIRED HCCS AND 30 CONTROL LIVERS). FINALLY, WE USED LUCIFERASE REPORTER AND COLONY FORMATION ASSAY TO CHECK THE EFFECT OF SOX1 IN HCC. RESULTS: PROMOTER METHYLATION OF SOX1 WAS SIGNIFICANTLY FREQUENT IN HCC CELL LINES AND CLINICAL HCCS, CIRRHOTIC LIVERS, BUT NOT IN CONTROL LIVERS (P < 0.0001). THERE IS A SIGNIFICANT CORRELATION BETWEEN DOWNREGULATION OF SOX1 EXPRESSION AND PROMOTER METHYLATION. QMSP RESULTS CONFIRMED THAT PROMOTER HYPERMETHYLATION OF SOX1 IS SIGNIFICANTLY MORE FREQUENT IN HCCS THAN CONTROL LIVERS (P < 0.0001). THE FREQUENCY OF SOX1 METHYLATION IN PATIENTS WITH SECRETED FRIZZLED-RELATED PROTEINS (SFRPS) METHYLATION IS SIGNIFICANTLY HIGHER THAN IN PATIENTS WITHOUT SFRPS METHYLATION (P < 0.0001). FURTHERMORE, ECTOPIC EXPRESSION OF SOX1 COULD SUPPRESS T-CELL FACTOR-DEPENDENT TRANSCRIPTIONAL ACTIVITY AND COLONY FORMATION NUMBER IN HCCS. CONCLUSIONS: CONCOMITANT EPIGENETIC SILENCING OF SOX1 AND SFRPS THROUGH PROMOTER HYPERMETHYLATION IS FREQUENT IN HCCS, AND THIS MIGHT CONTRIBUTE TO ABNORMAL ACTIVATION OF CANONICAL WNT SIGNAL PATHWAY.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
2  6770  46 [ABERRANT METHYLATION OF MULTIPLE GENES AND ITS CLINICAL IMPLICATION IN HEPATOCELLULAR CARCINOMA]. OBJECTIVE: TO INVESTIGATE THE METHYLATION FREQUENCIES OF MULTIPLE TUMOR SUPPRESSOR GENES (TSGS) IN HEPATOCELLULAR CARCINOMA (HCC) AND THE CLINICAL IMPLICATION OF ABERRANT DNA METHYLATION IN MOLECULAR CARCINOGENESIS OF HCC. METHODS: SIXTY SAMPLES OF HCC AND THE PAIRED ADJACENT LIVER TISSUE, 16 SAMPLES FROM POST-HEPATITIS CIRRHOTIC LIVERS, 5 FROM LIVERS WITH CHRONIC HEPATITIS AND 5 FROM NORMAL LIVERS WERE COLLECTED. EIGHT TSGS FREQUENTLY SILENCED BY HYPERMETHYLATION OF THEIR PROMOTERS IN VARIOUS TYPES OF DIGESTIVE TUMORS WERE SELECTED, INCLUDING APC, RASSF1A, P16, GSTP1, MGMT, DAPK, SOCS-1 AND RIZ1. THE STATUS OF PROMOTER METHYLATION IN THESE 8 GENES WAS INVESTIGATED USING METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. THE CLINICOPATHOLOGICAL DATA OF HCC WERE ALSO ANALYZED IN ORDER TO EVALUATE THE CLINICAL IMPLICATION OF ABERRANT METHYLATION IN HCC. RESULTS: METHYLATION OF THE 8 TSGS WAS QUITE FREQUENT IN HCC, WITH A METHYLATION RATE OF 95.0% IN RASSF1A, 90.0% IN APC, 73.3% IN GSTP1, 65.0% IN P16, 61.6% IN RIZ1 AND 60.0% IN MGMT. METHYLATION OF THE 6 GENES WAS MORE FREQUENT IN HCC THAN THAT IN ADJACENT TISSUES (P < 0.05). THE METHYLATION RATE OF MGMT, GSTP1 AND RIZ1 IN THE ADJACENT TISSUES WAS 41.6%, 40.0% AND 25.0%, RESPECTIVELY, SIGNIFICANTLY HIGHER THAN THAT IN CIRRHOTIC LIVER (P < 0.05). P16 METHYLATION WAS MORE FREQUENTLY OBSERVED IN HCC IN ELDERLY PATIENTS. THE FREQUENCY OF MGMT METHYLATION WAS TENDED TO BE HIGHER IN GIANT HCC THAN THAT IN THE OTHER TYPES OF HCC. PATIENTS WITH MGMT METHYLATION IN THE TUMOR WERE FOUND TO HAVE A SHORTER DISEASE FREE SURVIVAL. CONCLUSION: DIFFERENT FREQUENCY OF METHYLATION IN HEPATOCELLULAR CARCINOMAS, ADJACENT LIVER TISSUES AND CIRRHOTIC LIVERS IMPLIES THAT EPIGENETIC ALTERATION IN THE HEPATOCELLULAR CARCINOGENESIS MAY BE A GRADUALLY PROGRESSIVE PROCESS. METHYLATION STATUS OF MGMT, GSTP1 AND RIZ1 MAY BE PROMISING IN RISK ASSESSMENT OF HEPATOCELLULAR CARCINOMA AND IN EARLY DIAGNOSIS. FURTHERMORE, MGMT METHYLATION MIGHT BE ALSO USED AS A POTENTIAL PROGNOSTIC BIOMARKER FOR HEPATOCELLULAR CARCINOMA PATIENTS.	2008	
                                                                                                                                                                                                                                                                                                                                                               
3  4903  34 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
4  3825  40 INVESTIGATION OF CTNNB1 GENE MUTATIONS AND EXPRESSION IN HEPATOCELLULAR CARCINOMA AND CIRRHOSIS IN ASSOCIATION WITH HEPATITIS B VIRUS INFECTION. HEPATITIS B VIRUS (HBV), ALONG WITH HEPATITIS C VIRUS CHRONIC INFECTION, REPRESENTS A MAJOR RISK FACTOR FOR HEPATOCELLULAR CARCINOMA (HCC) DEVELOPMENT. HOWEVER, MOLECULAR MECHANISMS INVOLVED IN THE DEVELOPMENT OF HCC ARE NOT YET COMPLETELY UNDERSTOOD. RECENT STUDIES HAVE INDICATED THAT MUTATIONS IN CTNNB1 GENE ENCODING FOR BETA-CATENIN PROTEIN LEAD TO ABERRANT ACTIVATION OF THE WNT/ BETA-CATENIN PATHWAY. THE MUTATIONS IN TURN ACTIVATE SEVERAL DOWNSTREAM GENES, INCLUDING C-MYC, PROMOTING THE NEOPLASTIC PROCESS. THE PRESENT STUDY EVALUATED THE MUTATIONAL PROFILE OF THE CTNNB1 GENE AND EXPRESSION LEVELS OF CTNNB1 AND C-MYC GENES IN HBV-RELATED HCC, AS WELL AS IN CIRRHOTIC AND CONTROL TISSUES. MUTATIONAL ANALYSIS OF THE BETA-CATENIN GENE AND HBV GENOTYPING WERE CONDUCTED BY DIRECT SEQUENCING. EXPRESSION OF BETA-CATENIN AND C-MYC GENES WAS ASSESSED USING REAL-TIME PCR. AMONG THE HCC CASES, 18.1% SHOWED MISSENSE POINT MUTATION IN EXON 3 OF CTNNB1, MORE FREQUENTLY IN CODONS 32, 33, 38 AND 45. THE FREQUENCY OF MUTATION IN THE HOTSPOTS OF EXON 3 WAS SIGNIFICANTLY HIGHER IN NON-VIRAL HCCS (29.4%) RATHER THAN HBV-RELATED CASES (12.7%, P = 0.021). THE EXPRESSION OF BETA-CATENIN AND C-MYC GENES WAS FOUND UPREGULATED IN CIRRHOTIC TISSUES IN ASSOCIATION WITH HBV INFECTION. MUTATIONS AT BOTH PHOSPHORYLATION AND NEIGHBORING SITES WERE ASSOCIATED WITH INCREASED ACTIVITY OF THE WNT PATHWAY. THE RESULTS DEMONSTRATED THAT MUTATED BETA-CATENIN CAUSED ACTIVATION OF THE WNT PATHWAY, BUT THE RATE OF CTNNB1 GENE MUTATIONS WAS NOT RELATED TO HBV INFECTION. HBV FACTORS MAY DEREGULATE THE WNT PATHWAY BY CAUSING EPIGENETIC ALTERATIONS IN THE HBV-RELATED HCC.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
5  5349  40 RASSF1A AND DOK1 PROMOTER METHYLATION LEVELS IN HEPATOCELLULAR CARCINOMA, CIRRHOTIC AND NON-CIRRHOTIC LIVER, AND CORRELATION WITH LIVER CANCER IN BRAZILIAN PATIENTS. HEPATOCELLULAR CARCINOMA (HCC) IS THE SECOND MOST COMMON CAUSE OF CANCER MORTALITY WORLDWIDE. MOST CASES OF HCC ARE ASSOCIATED WITH CIRRHOSIS RELATED TO CHRONIC HEPATITIS B VIRUS OR HEPATITIS C VIRUS INFECTIONS. HYPERMETHYLATION OF PROMOTER REGIONS IS THE MAIN EPIGENETIC MECHANISM OF GENE SILENCING AND HAS BEEN INVOLVED IN HCC DEVELOPMENT. THE AIM OF THIS STUDY WAS TO DETERMINE WHETHER ABERRANT METHYLATION OF RASSF1A AND DOK1 GENE PROMOTERS IS ASSOCIATED WITH THE PROGRESSION OF LIVER DISEASE IN BRAZILIAN PATIENTS. METHYLATION LEVELS WERE MEASURED BY PYROSEQUENCING IN 41 (20 HCC, 9 CIRRHOTIC, AND 12 NON-CIRRHOTIC) LIVER TISSUE SAMPLES. MEAN RATES OF METHYLATION IN RASSF1A AND DOK1 WERE 16.2% AND 12.0% IN NON-CIRRHOTIC, 26.1% AND 19.6% IN CIRRHOTIC, AND 59.1% AND 56.0% IN HCC TISSUES, RESPECTIVELY, SHOWING A GRADUAL INCREASE ACCORDING TO THE PROGRESSION OF THE DISEASE, WITH SIGNIFICANTLY HIGHER LEVELS IN TUMOR TISSUES. IN ADDITION, HYPERMETHYLATION OF RASSF1A AND DOK1 WAS FOUND IN THE VAST MAJORITY (88%) OF THE HCC CASES. INTERESTINGLY, DOK1 METHYLATION LEVELS IN HCC SAMPLES WERE SIGNIFICANTLY HIGHER IN THE GROUP OF YOUNGER (<40 YEARS) PATIENTS, AND HIGHER IN MODERATELY DIFFERENTIATED THAN IN POORLY DIFFERENTIATED TUMORS (P < 0.05). OUR RESULTS REINFORCE THE HYPOTHESIS THAT HYPERMETHYLATION OF RASSF1A AND DOK1 CONTRIBUTES TO HEPATOCARCINOGENESIS AND IS ASSOCIATED TO CLINICOPATHOLOGICAL CHARACTERISTICS. RASSF1A AND DOK1 PROMOTER HYPERMETHYLATION MAY BE A VALUABLE BIOMARKER FOR EARLY DIAGNOSIS OF HCC AND A POTENTIAL MOLECULAR TARGET FOR EPIGENETIC-BASED THERAPY.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
6  6208  34 THE INTERACTION BETWEEN MICRORNA-152 AND DNA METHYLTRANSFERASE-1 AS AN EPIGENETIC PROGNOSTIC BIOMARKER IN HCV-INDUCED LIVER CIRRHOSIS AND HCC PATIENTS. THE NECESSITY FOR EARLY DETECTION AND HENCE IMPROVING THE OUTCOME OF TREATMENT OF HEPATOCELLULAR CARCINOMA (HCC) IS CRITICAL ESPECIALLY IN HEPATITIS C VIRUS (HCV)-GENOTYPE 4 INDUCED CASES. IN OUR CURRENT WORK, WE EXAMINED THE MIRNA-152 AND DNMT-1 EXPRESSION IN CHRONIC LIVER DISEASE (CLD) DUE TO HCV GENOTYPE 4 INFECTION WITH/WITHOUT CIRRHOSIS AND HCC PATIENTS AS AN ATTEMPT TO EVALUATE THE POTENTIAL BENEFITS OF THESE NEW CIRCULATING, NONINVASIVE, PROGNOSTIC, EPIGENETIC MARKERS FOR LIVER CIRRHOSIS AND CARCINOGENESIS OF EGYPTIAN PATIENTS. EIGHTY SUBJECTS WERE INCLUDED IN THIS STUDY, DIVIDED INTO TWO GROUPS; GROUP I (40 PATIENTS) WERE CLASSIFIED INTO SUBGROUP IA (CLD WITHOUT CIRRHOSIS, N = 18) AND SUBGROUP IB (CLD WITH CIRRHOSIS, N = 22), GROUP II (CLD PATIENTS WITH HCC, N = 20), AND CONTROL (HEALTHY VOLUNTEER, N = 20). THE EXPRESSION OF MIRNA-152 AND DNMT-1 GENES WERE ANALYZED USING REAL-TIME PCR. MIRNA-152 SHOWED A PERSISTENT AND SIGNIFICANT DOWNREGULATION IN ALL DISEASED GROUPS, WHICH WAS IN CONSISTENCE WITH THE PROGRESSION OF THE DISEASE TOWARD THE HCC STAGE. DNMT-1 SHOWED UPREGULATION IN ALL DISEASED GROUPS WHEN COMPARED TO CONTROL AND SUBGROUP IA. THE MIRNA-152 WAS SHOWN TO CORRELATE INVERSELY WITH DNMT-1 IN SUBGROUP IA, IB AND GROUP II (R = -0.557, P < 0.01), (R = -0.850, P < 0.001) AND (R = -0.544, P < 0.02) RESPECTIVELY. IN ADDITION, MIRNA-152 AND DNMT-1 SHOWED A DIAGNOSTIC ABILITY TO DISCRIMINATE BETWEEN CASES OF CIRRHOSIS AND HCC AGAINST CLD WITHOUT CIRRHOSIS (P < 0.01), WHILE DNMT-1 DID NOT, EXCEPT BETWEEN HCC AND CIRRHOTIC CASES. FURTHERMORE, BOTH GENES CAN BE CONSIDERED AS PREDICTOR AND PROGNOSTIC PARAMETERS FOR CIRRHOSIS (OR = 1.041, P = 0.043) AND (OR = 1.039, P = 0.04) RESPECTIVELY, WHILE MIRNA-152 ALONE IS PROVED AS A PROGNOSTIC MARKER FOR HCC (OR = 1.003, P = 0.044). FINALLY, THE PERSISTENT REVERSE CORRELATION BETWEEN MIRNA-152 WITH DNMT-1 PROMPTS THEIR USE AS NONINVASIVE PROGNOSTIC BIOMARKERS FOR HCV INDUCED LIVER CIRRHOSIS AND HCC IN HCV GENOTYPE 4 PATIENTS.	2020	
                                                                                                                                                                                                                                                                                                                                                                              
7  1342  43 DETECTING ABNORMAL METHYLATION OF TUMOR SUPPRESSOR GENES GSTP1, P16, RIZ1, AND RASSF1A IN HEPATOCELLULAR CARCINOMA AND ITS CLINICAL SIGNIFICANCE. HEPATOCELLULAR CARCINOMA (HCC) HAS A HIGH RATE OF MORTALITY. FURTHER STUDIES INTO EPIGENETIC CHANGES IN HCC, PARTICULARLY THE ABNORMAL METHYLATION OF TUMOR SUPPRESSOR GENES (TSGS), ARE REQUIRED, SINCE THESE CHANGES MAY PROVIDE NOVEL BIOMARKERS FOR EARLY SCREENING AND DIAGNOSIS OF HCC. BY USING METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP), THE PRESENT STUDY DETECTED THE METHYLATION STATUS IN THE PROMOTER REGION OF 4 CANDIDATE TSGS, GSTP1, P16, RIZ1, AND RASSF1A, RESPECTIVELY, IN 35 PAIRED HCC AND TUMOR-ADJACENT LIVER TISSUES IN ADDITION TO 20 NORMAL LIVER TISSUES. THEIR EFFECT ON THE INITIATION AND PROGRESSION OF HCC WAS ALSO INVESTIGATED BY ANALYZING THE CLINICOPATHOLOGICAL DATA. THE RESULTS OF THE PRESENT STUDY REVEALED THAT THE METHYLATION LEVEL OF RIZ1 AND GSTP1 GENES IN HCC WAS SIGNIFICANTLY INCREASED COMPARED WITH THAT IN THE ADJACENT TISSUES (P<0.01) AND THE NORMAL LIVER TISSUES (P<0.01). THE METHYLATION FREQUENCY OF P16 AND RASSF1A GENES WAS NOT SIGNIFICANTLY INCREASED COMPARED WITH THAT OBSERVED IN THE ADJACENT TISSUES (P>0.05) BUT WAS SIGNIFICANTLY INCREASED COMPARED WITH THE NORMAL TISSUES (P<0.01). IN HCC TISSUES, THE METHYLATION FREQUENCY OF THE GSTP1 GENE IN TUMORS WITH CAPSULAR INVASION WAS SIGNIFICANTLY INCREASED COMPARED WITH THAT IN TUMORS WITHOUT CAPSULAR INVASION (P<0.05). THE METHYLATION FREQUENCY OF P16 GENE IN HEPATITIS B SURFACE ANTIGEN (HBSAG)-POSITIVE HCC PATIENTS WAS SIGNIFICANTLY INCREASED COMPARED WITH THAT IN HBSAG-NEGATIVE PATIENTS (P<0.05). THE METHYLATION STATUS OF RIZ1 AND RASSF1A GENES WAS NOT SIGNIFICANTLY CORRELATED WITH THE CLINICOPATHOLOGICAL DATA (P>0.05). PREVIOUS STUDIES HAVE DEMONSTRATED THAT THE METHYLATION STATUS OF RIZ1 AND GSTP1 GENES IS HCC-SPECIFIC, AND THUS MAY BE USED AS A BIOMARKER TO ASSIST THE CLINICAL DIAGNOSIS OF HCC. WHILE THE METHYLATION OF GSTP1 GENE PROMOTER MAY ASSOCIATE WITH THE INVASIVENESS OF HCC, CHRONIC HEPATITIS B VIRUS INFECTION MAY BE THE CAUSE OF METHYLATION-INDUCED P16 INACTIVATION.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                   
8  5338  45 QUANTITATIVE EVALUATION OF RASSF1A METHYLATION IN THE NON-LESIONAL, REGENERATIVE AND NEOPLASTIC LIVER. BACKGROUND: EPIGENETIC CHANGES DURING AGEING AND THEIR RELATIONSHIP WITH CANCER ARE UNDER THE FOCUS OF INTENSE RESEARCH. RASSF1A AND NORE1A ARE NOVEL GENES ACTING IN CONCERT IN THE PROAPOPTOTIC PATHWAY OF THE RAS SIGNALLING. WHILE NORE1A HAS NOT BEEN PREVIOUSLY INVESTIGATED IN THE HUMAN LIVER, RECENT REPORTS HAVE SUGGESTED THAT RASSF1A IS FREQUENTLY EPIGENETICALLY METHYLATED NOT ONLY IN HCC BUT ALSO IN THE CIRRHOTIC LIVER. METHODS: TO ADDRESS WHETHER EPIGENETIC CHANGES TAKE PLACE IN CONNECTION TO AGE AND/OR TO THE UNDERLYING DISEASE, WE INVESTIGATED RASSF1A AND NORE1A GENE PROMOTER METHYLATION BY CONVENTIONAL METHYLATION SPECIFIC PCR AND REAL-TIME MSP IN A SERIES OF HEPATITIC AND NON-HEPATITIC LIVERS HARBORING REGENERATIVE/HYPERPLASTIC (CIRRHOSIS/FOCAL NODULAR HYPERPLASIA), DYSPLASTIC (LARGE REGENERATIVE, LOW AND HIGH GRADE DYSPLASTIC NODULES) AND NEOPLASTIC (HEPATOCELLULAR ADENOMA AND CARCINOMA) GROWTHS. RESULTS: IN THE HEPATITIC LIVER (CHRONIC HEPATITIC/CIRRHOSIS, HEPATOCELLULAR NODULES AND HCC) WE FOUND WIDESPREAD RASSF1A GENE PROMOTER METHYLATION WITH A METHYLATION INDEX THAT INCREASED FROM REGENERATIVE CONDITIONS (CIRRHOSIS) TO HEPATOCELLULAR NODULES (P < 0.01) TO HCC (P < 0.001). IN THE NON-HEPATITIC LIVER A CONSISTENT PATTERN OF GENE METHYLATION WAS ALSO FOUND IN BOTH LESIONAL (FOCAL NODULAR HYPERPLASIA AND HEPATOCELLULAR ADENOMA) AND NON-LESIONAL TISSUE. SPECIFICALLY, HEPATOCELLULAR ADENOMAS (HA) SHOWED A METHYLATION INDEX SIGNIFICANTLY HIGHER THAN THAT DETECTED IN FOCAL NODULAR HYPERPLASIA (FNH) (P < 0.01) AND IN NON-LESIONAL TISSUE (P < 0.001). IN NON-LESIONAL LIVER ALSO THE METHYLATION INDEX GRADUALLY INCREASED BY AGEING (P = 0.002), SUGGESTING A PROGRESSIVE SPREADING OF METHYLATED CELLS OVER TIME. AS OPPOSED TO RASSF1A GENE PROMOTER METHYLATION, NORE1A GENE WAS NEVER FOUND EPIGENETICALLY ALTERATED IN BOTH HEPATITIC AND NON-HEPATITIC LIVER. CONCLUSION: WE HAVE SHOWN THAT IN NON-LESIONAL, REGENERATIVE AND NEOPLASTIC LIVER THE RASSF1A GENE IS INCREASINGLY METHYLATED, THAT THIS CONDITION TAKES PLACE AS AN AGE-RELATED PHENOMENON AND THAT THE EARLY SETTING AND SPREADING OVER TIME OF AN EPIGENETICALLY METHYLATED HEPATOCYTE SUBPOPULATION, MIGHT BE RELATED TO LIVER TUMORIGENESIS.	2006	
                                                                                                                                                                                                          
9  6747  38 WHOLE GENOME 5'-METHYLCYTOSINE LEVEL QUANTIFICATION IN CIRRHOTIC HCV-INFECTED EGYPTIAN PATIENTS WITH AND WITHOUT HEPATOCELLULAR CARCINOMA. DNA METHYLATION IS AN EPIGENETIC MECHANISM USED BY CELLS TO CONTROL GENE EXPRESSION. DNA METHYLATION IS A COMMONLY USED EPIGENETIC SIGNALING TOOL THAT CAN HOLD GENES IN THE "OFF" POSITION. CHRONIC INFECTION WITH HEPATITIS C VIRUS (HCV) IS CONSIDERED A MAJOR RISK FOR CHRONIC LIVER IMPAIRMENT. IT IS THE MOST COMMON LEADING CAUSE OF HCC. THE PRESENT WORK IS AIMED AT STUDYING WHOLE GENOME 5'-METHYLCYTOSINE LEVELS IN CIRRHOTIC HCV-INFECTED EGYPTIAN PATIENTS. IN THE PRESENT STUDY, 120 EGYPTIAN ADULTS WERE INCLUDED. THEY WERE DIVIDED INTO TWO GROUPS: GROUP CAPITAL I, UKRAINIAN (40 APPARENTLY HEALTHY CONTROL SUBJECTS) AND GROUP CAPITAL I, UKRAINIANCAPITAL I, UKRAINIAN (80 HCV-INFECTED PATIENTS). FURTHERMORE, GROUP II WAS SUBDIVIDED INTO 2 SUBGROUPS ACCORDING TO THE PRESENCE OF HCC IN HCV-INFECTED SUBJECTS. TO ALL STUDIED SUBJECTS, THE LEVEL OF 5-MC% WAS MEASURED IN PERIPHERAL BLOOD. IN THE PRESENT STUDY, THE MEDIAN OF 5'-METHYLCYTOSINE% IN THE CONTROL GROUP (GROUP I) WAS 2.5, IN THE HCV GROUP (GROUP IIA) WAS 2.45, AND IN THE HCC GROUP (GROUP II B) WAS 2.25. A STEPWISE DECREASE IN 5'-METHYLCYTOSINE% FROM THE CONTROL (GROUP I) TOWARD HCC (GROUP IIB) WAS OBSERVED, TAKING INTO CONSIDERATION THAT THE STEPWISE GLOBAL HYPOMETHYLATION WAS NOT STATISTICALLY SIGNIFICANT (P = 0.811). THERE WAS A NEGATIVE CORRELATION BETWEEN ALT AND 5'-METHYLCYTOSINE% (P = -0.029). FROM THIS STUDY, WE CAN CONCLUDE THAT GLOBAL DNA 5'-METHYLCYTOSINE% DOES NOT DIFFER IN HCV-INFECTED CIRRHOTIC PATIENTS AND HCC PATIENTS WHEN COMPARED TO NORMAL CONTROLS. CONSECUTIVELY, WE HAD CONCLUDED THAT THERE IS NO IMPACT OF 5'-METHYLCYTOSINE% ON THE DEVELOPMENT OF LIVER CIRRHOSIS OR HCC. MOREOVER, THE NEGATIVE CORRELATION BETWEEN 5'-METHYLCYTOSINE% AND SERUM ALT LEVEL DENOTES A TREND OF DECREASE IN 5'-METHYLCYTOSINE% WITH MORE LIVER DAMAGE.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
10  1393  40 DIAGNOSTIC VALUE OF THE HYPOMETHYLATION OF THE WISP1 PROMOTER IN PATIENTS WITH HEPATOCELLULAR CARCINOMA ASSOCIATED WITH HEPATITIS B VIRUS. WNT1-INDUCIBLE SIGNALING PATHWAY PROTEIN 1 (WISP1) REGULATES CELL PROLIFERATION, DIFFERENTIATION, ADHESION, MIGRATION AND SURVIVAL. ABNORMAL WISP1 EXPRESSION IS ASSOCIATED WITH THE CARCINOGENESIS OF HEPATOCELLULAR CARCINOMA (HCC). ABERRANT DNA METHYLATION IS ONE OF THE MAJOR EPIGENETIC ALTERATIONS IN HCC. HOWEVER, THE METHYLATION STATUS OF THE WISP1 PROMOTER IS STILL UNCLEAR. WE THEREFORE AIMED TO DETERMINE THE METHYLATION STATUS OF THE WISP1 PROMOTER AND EVALUATE ITS CLINICAL VALUE IN HCC. THE STUDY ENROLLED 251 PARTICIPANTS, INCLUDING 123 PARTICIPANTS WITH HCC, 90 PARTICIPANTS WITH CHRONIC HEPATITIS B (CHB) AND 38 HEALTHY CONTROLS (HCS). WISP1 METHYLATION STATUS, MRNA LEVELS AND PLASMA SOLUBLE WISP1 WERE DETECTED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP), QUANTITATIVE REAL-TIME PCR (RT-QPCR) AND ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA), RESPECTIVELY. WE FOUND THAT THE METHYLATION FREQUENCY OF WISP1 IN PATIENTS WITH HCC WAS SIGNIFICANTLY LOWER THAN THAT IN PATIENTS WITH CHB AND HCS, WHILE THE RELATIVE EXPRESSION LEVELS OF WISP1 MRNA WERE MARKEDLY HIGHER IN PATIENTS WITH HCC THAN IN PATIENTS WITH CHB AND HCS. FURTHERMORE, THE PLASMA SOLUBLE WISP1 IN PATIENTS WITH HCC WAS OBVIOUSLY LOWER THAN IN THAT IN PATIENTS WITH CHB AND HCS. ALPHA-FETOPROTEIN (AFP) IS A WIDELY RECOGNIZED BIOMARKER TO DIAGNOSE HCC WHICH LACKS ENOUGH SENSITIVITY AND SPECIFICITY. WISP1 PROMOTER METHYLATION STATUS COMBINED WITH AFP SIGNIFICANTLY IMPROVED THE DIAGNOSTIC ABILITY IN DISCRIMINATING HCC FROM CHB COMPARED WITH AFP OR WISP1 METHYLATION STATUS ALONE. IN CONCLUSION, HYPOMETHYLATION OF THE WISP1 GENE PROMOTER MAY SERVE AS A NONINVASIVE BIOMARKER FOR DETECTING HBV-ASSOCIATED HCC.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
11   153  41 ABERRANT METHYLATION OF MULTIPLE TUMOR SUPPRESSOR GENES IN AGING LIVER, CHRONIC HEPATITIS, AND HEPATOCELLULAR CARCINOMA. ABERRANT DNA METHYLATION IS AN IMPORTANT EPIGENETIC ALTERATION IN HEPATOCELLULAR CARCINOMA (HCC). HOWEVER, THE MOLECULAR PROCESSES UNDERLYING THE METHYLATOR PHENOTYPE AND THE CONTRIBUTION OF HEPATITIS VIRUSES ARE POORLY UNDERSTOOD. THE CURRENT STUDY IS A COMPREHENSIVE METHYLATION ANALYSIS OF HUMAN LIVER TISSUE SPECIMENS. A TOTAL OF 176 LIVER TISSUES, INCLUDING 77 PAIRS OF HCCS AND MATCHING NONCANCEROUS LIVER AND 22 NORMAL LIVERS, WERE ANALYZED FOR METHYLATION. METHYLATION OF 19 EPIGENETIC MARKERS WAS QUANTIFIED, AND THE RESULTS WERE CORRELATED WITH DIFFERENT DISEASE STATES AND THE PRESENCE OR ABSENCE OF HEPATITIS B VIRUS (HBV) AND HEPATITIS C VIRUS (HCV) INFECTIONS. BASED ON METHYLATION PROFILES, THE 19 LOCI WERE CATEGORIZED INTO 3 GROUPS. NORMAL LIVER TISSUES SHOWED METHYLATION PRIMARILY IN GROUP 1 LOCI (HIC-1, CASP8, GSTP1, SOCS1, RASSF1A, P16, APC), WHICH WAS SIGNIFICANTLY HIGHER THAN GROUP 2 (CDH1, RUNX3, RIZ1, SFRP2, MINT31) AND GROUP 3 MARKERS (COX2, MINT1, CACNA1G, RASSF2, MINT2, REPRIMO, DCC) (P < 0.0001). NONCANCEROUS LIVERS DEMONSTRATED INCREASED METHYLATION IN BOTH GROUP 1 AND GROUP 2 LOCI. METHYLATION WAS SIGNIFICANTLY MORE ABUNDANT IN HCV-POSITIVE LIVERS COMPARED WITH NORMAL LIVER TISSUES. CONVERSELY, HCC SHOWED FREQUENT METHYLATION AT EACH LOCUS INVESTIGATED IN ALL 3 GROUPS. HOWEVER, THE GROUP 3 LOCI SHOWED MORE DENSE AND FREQUENT METHYLATION IN HCV-POSITIVE CANCERS COMPARED WITH BOTH HBV-POSITIVE CANCERS AND VIRUS-NEGATIVE CANCERS (P < 0.0001). CONCLUSION: METHYLATION IN HCC IS FREQUENT BUT OCCURS IN A GENE-SPECIFIC AND DISEASE-SPECIFIC MANNER. METHYLATION PROFILING ALLOWED US TO DETERMINE THAT ABERRANT METHYLATION IS COMMONLY PRESENT IN NORMAL AGING LIVERS, AND SEQUENTIALLY PROGRESSES WITH ADVANCING STAGES OF CHRONIC VIRAL INFECTION. FINALLY, OUR DATA PROVIDE EVIDENCE THAT HCV INFECTION MAY ACCELERATE THE METHYLATION PROCESS AND SUGGESTS A CONTINUUM OF INCREASING METHYLATION WITH PERSISTENT VIRAL INFECTION AND CARCINOGENESIS IN THE LIVER.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                
12   817  40 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
13  1617  40 DNA METHYLTRANSFERASE EXPRESSION AND DNA METHYLATION IN HUMAN HEPATOCELLULAR CARCINOMA AND THEIR CLINICOPATHOLOGICAL CORRELATION. ABERRANT DNA METHYLATION ON CPG ISLANDS IS ONE OF THE MOST CONSISTENT EPIGENETIC CHANGES IN HUMAN CANCERS, AND THE METHYLATION PROCESS IS CATALYZED BY DNA METHYLTRANSFERASE (DNMT). WE EVALUATED I) THE MRNA LEVELS OF THREE DNMTS; DNMT1, DNMT3A AND DNMT3B, IN 25 HEPATOCELLULAR CARCINOMAS (HCCS), IN THEIR CORRESPONDING NON-CANCEROUS LIVER TISSUES AND IN 7 NORMAL LIVERS BY USING REAL-TIME REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION; II) NUCLEAR EXPRESSION OF DNMT1 AND DNMT3A PROTEINS IN THE HCCS BY IMMUNOHISTOCHEMISTRY, III) THE METHYLATION STATUS OF 5 GENES; P16, P15, E-CADHERIN, HIC-1 AND RASSF1A IN THE SAME TISSUES, AND IV) THE RELATIONSHIPS BETWEEN THE ABOVE RESULTS AND THE CLINICOPATHOLOGICAL CHARACTERISTICS, INCLUDING PROGNOSIS. THE DIFFERENCES IN MRNA EXPRESSION LEVELS FOR DNMT1, DNMT3A AND DNMT3B WERE STATISTICALLY SIGNIFICANT BETWEEN HCC AND NORMAL LIVERS (P<0.001), HCC AND CHRONIC HEPATITIS (P<0.001) AND HCC AND CIRRHOSIS (P<0.001). AN INCREASE IN MRNA EXPRESSION LEVELS OF >4-FOLD FOR DNMT3B IN HCCS WAS SIGNIFICANTLY ASSOCIATED WITH A POORER OVERALL SURVIVAL (P=0.027) AND SHORTER METASTASIS-FREE SURVIVAL (P=0.0299). A POORER RECURRENCE-FREE SURVIVAL WAS NOTED IN HCCS WITH A >4-FOLD INCREASE IN DNMT3A MRNA (P=0.0120). THE AVERAGE NUMBERS OF METHYLATED GENES WERE 0, 1.27, 1.38 AND 2.72 FOR NORMAL LIVERS, CHRONIC HEPATITIS, CIRRHOSIS AND HCCS, RESPECTIVELY, AND THIS PROGRESSIVE INCREASE FROM NORMAL LIVERS TO CHRONIC HEPATITIS/CIRRHOSIS THROUGH HCC MAY SUGGEST THAT TUMOR SUPPRESSOR GENE METHYLATION IS AN EARLY EVENT IN HEPATOCARCINOGENESIS. THESE RESULTS FIRST SUGGEST THAT HEPATOCARCINOGENESIS INVOLVES AN INCREASED EXPRESSION OF DNMT1, DNMT3A AND DNMT3B MRNA AND A PROGRESSIVE INCREASE IN THE NUMBER OF METHYLATED GENES FROM NORMAL LIVER, CHRONIC HEPATITIS/CIRRHOSIS TO HCC AND SECONDLY THAT AN INCREASE IN THE DNMT3A AND DNMT3B MRNA LEVELS IN HCCS RELATIVE TO THEIR NON-CANCEROUS TISSUES MAY BE A PREDICTOR OF POOR SURVIVAL.	2007	
                                                                                                                                                                                                                                                                                                                                                                                                                                                    
14  4246  39 METHYLATION STATUS OF THE T-CADHERIN GENE PROMOTOR IN PERIPHERAL BLOOD MONONUCLEAR CELLS IS ASSOCIATED WITH HBV-RELATED HEPATOCELLULAR CARCINOMA PROGRESSION. DNA METHYLATION IS ONE OF THE EPIGENETIC MECHANISMS TO REGULATE GENE EXPRESSION AND FREQUENTLY OCCURS IN HUMAN CANCER CELLS. T-CADHERIN (CDH13) IS A NEW MEMBER OF THE CADHERIN SUPERFAMILY AND POSSESSES MULTIPLE FUNCTIONS. OUR STUDY INCLUDED 26 NORMAL CONTROLS (NCS), 65 CHRONIC HEPATITIS B PATIENTS (CHB), 14 LIVER CIRRHOSIS PATIENTS (LC) AND 157 HEPATOCELLULAR CARCINOMA PATIENTS (HCC). WE MAINLY FOCUSED ON THE MRNA EXPRESSION AND METHYLATION STATUS OF CDH13 IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS), WHICH WERE DETECTED BY SEMI-QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (RT-QPCR) AND METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) RESPECTIVELY. THE CDH13 MRNA LEVEL WAS LOWER IN HCC, ESPECIALLY IN EARLY-STAGE OF HCC THAN IN NCS AND CHB GROUPS (P < 0.05). METHYLATION FREQUENCY OF THE CDH13 PROMOTER WAS SIGNIFICANTLY HIGHER IN HCC PATIENTS THAN IN THE NCS AND CHB GROUPS (67.52 % VS 0.00 %, P < 0.001, 67.52 % VS 52.31 %, P < 0.05, RESPECTIVELY). CDH13 MRNA LEVEL WAS SIGNIFICANTLY AND RELATIVELY LOWER IN METHYLATED GROUPS THAN IN UNMETHYLATED GROUPS AMONG THE WHOLE PARTICIPANTS. THE METHYLATION LEVEL OF CDH13 PROMOTER IN HCC MIGHT BE INFLUENCED OR PARTLY INFLUENCED BY SOME CRITICAL FACTORS SUCH AS TBIL, ALB AND AFP (P < 0.05). AS AN IMPORTANT FACTOR IN SIGNALING PATHWAY REGULATING BY CDH13 TO PROMOTE CARCINOGENESIS, JNK LEVEL WAS SIGNIFICANTLY HIGHER IN HCC WHICH HAD A HIGHER METHYLATION FREQUENCY THAN IN NCS, CHB AND LC (P < 0.05). FURTHERMORE, THE COMBINATION OF THE METHYLATED CDH13 LEVEL AND AFP LEVEL SHOWED A BETTER SCORE: AUC = 0.796 (SE = 0.031, 95 %CI 0.735-0.857; P < 0.001) IN MALE AND AUC = 0.832 (SE = 0.057, 95 %CI 0.721-0.944; P < 0.001) IN FEMALE COMPARED TO AFP ALONE FOR DIAGNOSING HCC FROM NCS, CHB AND LC. THE METHYLATION OF CDH13 PROMOTER WAS AN INDEPENDENT PREDICTOR FOR ASSESSING THE PROGNOSIS OF HCC PATIENTS (R=-1.378 P < 0.05). IN CONCLUSION, HYPERMETHYLATION OF CDH13 IN PBMCS WAS ASSOCIATED WITH THE UNDEREXPRESSION OF MRNA AND THE HIGH RISK OF HCC. THE METHYLATION STATUS OF THE CDH13 PROMOTER IN PBMCS WAS A POTENTIAL NONINVASIVE BIOMARKER TO PREDICT THE PROGNOSIS OF HCC PATIENTS.	2020	
                                                                                                                                                                                                                                
15  2844  48 FREQUENT EPIGENETIC INACTIVATION OF SFRP GENES IN HEPATOCELLULAR CARCINOMA. BACKGROUND: ACTIVATION OF THE WNT SIGNALING PATHWAY IS FREQUENTLY OBSERVED IN HEPATOCELLULAR CARCINOMA (HCC), THOUGH MUTATION OF THREE OF ITS COMPONENTS, CTNNB1, AXIN1, AND AXIN2, IS OBSERVED SUBSTANTIALLY LESS OFTEN. METHODS: WE EXAMINED THE RELATIONSHIP BETWEEN WNT SIGNALING AND EPIGENETIC ALTERATION OF SECRETED FRIZZLED-RELATED PROTEIN (SFRP) GENES IN HCC. RESULTS: WE FREQUENTLY DETECTED THE ACTIVE FORM OF BETA-CATENIN AND ACCUMULATION OF NUCLEAR BETA-CATENIN IN LIVER CANCER CELL LINES. WE DETECTED METHYLATION OF SFRP FAMILY GENES IN LIVER CANCER CELL LINES (SFRP1, 9/12, 75%; SFRP2, 7/12, 58%; SFRP4, 3/12, 25%; SFRP5, 7/12, 58%) AND PRIMARY HCCS (SFRP1, 9/19, 47%; SFRP2, 12/19, 63%; SFRP5, 8/19, 42%), THOUGH METHYLATION OF SFRP4 WAS NOT FOUND IN PRIMARY HCCS. SFRP METHYLATION ALSO WAS DETECTED IN HEPATITIS B OR C VIRUS-ASSOCIATED CHRONIC HEPATITIS (SFRP1, 6/37, 16%; SFRP2, 14/37, 38%; SFRP5, 5/37, 14%) AND LIVER CIRRHOSIS (SFRP1, 10/28, 36%; SFRP2, 9/28, 32%; SFRP5, 3/28, 11%), SUGGESTING THAT METHYLATION OF THESE GENES IS AN EARLY EVENT IN LIVER CARCINOGENESIS. ECTOPIC EXPRESSION OF SFRPS DOWNREGULATED T-CELL FACTOR/LYMPHOCYTE ENHANCER FACTOR (TCF/LEF) TRANSCRIPTIONAL ACTIVITY IN LIVER CANCER CELLS, WHILE OVEREXPRESSION OF A BETA-CATENIN MUTANT AND DEPLETION OF SFRP1 USING SIRNA SYNERGISTICALLY UPREGULATED TCF/LEF TRANSCRIPTIONAL ACTIVITY. CONCLUSIONS: OUR RESULTS CONFIRM THE FREQUENT METHYLATION AND SILENCING OF WNT ANTAGONIST GENES IN HCC, AND SUGGEST THAT THEIR LOSS OF FUNCTION CONTRIBUTES TO ACTIVATION OF WNT SIGNALING DURING HEPATOCARCINOGENESIS.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
16  3190  43 HCV-INDUCED EPIGENETIC CHANGES ASSOCIATED WITH LIVER CANCER RISK PERSIST AFTER SUSTAINED VIROLOGIC RESPONSE. BACKGROUND & AIMS: CHRONIC HEPATITIS C VIRUS (HCV) INFECTION IS AN IMPORTANT RISK FACTOR FOR HEPATOCELLULAR CARCINOMA (HCC). DESPITE EFFECTIVE ANTIVIRAL THERAPIES, THE RISK FOR HCC IS DECREASED BUT NOT ELIMINATED AFTER A SUSTAINED VIROLOGIC RESPONSE (SVR) TO DIRECT-ACTING ANTIVIRAL (DAA) AGENTS, AND THE RISK IS HIGHER IN PATIENTS WITH ADVANCED FIBROSIS. WE INVESTIGATED HCV-INDUCED EPIGENETIC ALTERATIONS THAT MIGHT AFFECT RISK FOR HCC AFTER DAA TREATMENT IN PATIENTS AND MICE WITH HUMANIZED LIVERS. METHODS: WE PERFORMED GENOME-WIDE CHIPMENTATION-BASED CHIP-SEQ AND RNA-SEQ ANALYSES OF LIVER TISSUES FROM 6 PATIENTS WITHOUT HCV INFECTION (CONTROLS), 18 PATIENTS WITH CHRONIC HCV INFECTION, 8 PATIENTS WITH CHRONIC HCV INFECTION CURED BY DAA TREATMENT, 13 PATIENTS WITH CHRONIC HCV INFECTION CURED BY INTERFERON THERAPY, 4 PATIENTS WITH CHRONIC HEPATITIS B VIRUS INFECTION, AND 7 PATIENTS WITH NONALCOHOLIC STEATOHEPATITIS IN EUROPE AND JAPAN. HCV-INDUCED EPIGENETIC MODIFICATIONS WERE MAPPED BY COMPARATIVE ANALYSES WITH MODIFICATIONS ASSOCIATED WITH OTHER LIVER DISEASE ETIOLOGIES. UPA/SCID MICE WERE ENGRAFTED WITH HUMAN HEPATOCYTES TO CREATE MICE WITH HUMANIZED LIVERS AND GIVEN INJECTIONS OF HCV-INFECTED SERUM SAMPLES FROM PATIENTS; MICE WERE GIVEN DAAS TO ERADICATE THE VIRUS. PATHWAYS ASSOCIATED WITH HCC RISK WERE IDENTIFIED BY INTEGRATIVE PATHWAY ANALYSES AND VALIDATED IN ANALYSES OF PAIRED HCC TISSUES FROM 8 PATIENTS WITH AN SVR TO DAA TREATMENT OF HCV INFECTION. RESULTS: WE FOUND CHRONIC HCV INFECTION TO INDUCE SPECIFIC GENOME-WIDE CHANGES IN H3K27AC, WHICH CORRELATED WITH CHANGES IN EXPRESSION OF MRNAS AND PROTEINS. THESE CHANGES PERSISTED AFTER AN SVR TO DAAS OR INTERFERON-BASED THERAPIES. INTEGRATIVE PATHWAY ANALYSES OF LIVER TISSUES FROM PATIENTS AND MICE WITH HUMANIZED LIVERS DEMONSTRATED THAT HCV-INDUCED EPIGENETIC ALTERATIONS WERE ASSOCIATED WITH LIVER CANCER RISK. COMPUTATIONAL ANALYSES ASSOCIATED INCREASED EXPRESSION OF SPHK1 WITH HCC RISK. WE VALIDATED THESE FINDINGS IN AN INDEPENDENT COHORT OF PATIENTS WITH HCV-RELATED CIRRHOSIS (N = 216), A SUBSET OF WHICH (N = 21) ACHIEVED VIRAL CLEARANCE. CONCLUSIONS: IN AN ANALYSIS OF LIVER TISSUES FROM PATIENTS WITH AND WITHOUT AN SVR TO DAA THERAPY, WE IDENTIFIED EPIGENETIC AND GENE EXPRESSION ALTERATIONS ASSOCIATED WITH RISK FOR HCC. THESE ALTERATIONS MIGHT BE TARGETED TO PREVENT LIVER CANCER IN PATIENTS TREATED FOR HCV INFECTION.	2019	

17  4220  40 METHYLATED CYSTEINE DIOXYGENASE-1 GENE PROMOTER IN THE SERUM IS A POTENTIAL BIOMARKER FOR HEPATITIS B VIRUS-RELATED HEPATOCELLULAR CARCINOMA. HEPATOCELLULAR CARCINOMA (HCC) IS THE THIRD LEADING CAUSE OF CANCER-RELATED MORTALITY WORLDWIDE. EPIGENETIC ANALYSIS HAS ATTRACTED INCREASING ATTENTION IN THE MOLECULAR DIAGNOSIS OF HCC. CYSTEINE DIOXYGENASE 1 (CDO1) IS A KEY ENZYME IN THE TAURINE BIOSYNTHETIC PATHWAY AND CONVERTS CYSTEINE TO CYSTEINE SULFINATE. THE CDO1 GENE IS A TUMOR SUPPRESSOR GENE AND IS USUALLY SILENCED BY THE METHYLATION OF ITS PROMOTER IN CARCINOGENESIS. IN THIS STUDY, WE EVALUATED WHETHER THE METHYLATION STATUS OF CDO1 GENE PROMOTER IS OF DIAGNOSTIC VALUE FOR HEPATITIS B VIRUS (HBV)-RELATED HCC. THE CDO1 PROMOTER METHYLATION STATUS WAS DETERMINED IN SERUM SAMPLES USING METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) IN A COHORT OF 123 PATIENTS WITH HBV-RELATED HCC, 28 WITH LIVER CIRRHOSIS (LC), 29 WITH CHRONIC HEPATITIS B (CHB) AND 20 HEALTHY CONTROLS. THE FREQUENCY OF THE CDO1 PROMOTER METHYLATION IN HBV-RELATED HCC (42.3%) WAS SIGNIFICANTLY HIGHER THAN THAT IN LC (14.3%), CHB (6.9%) AND HEALTHY CONTROLS (0%) (P = 0.006; P < 0.0001; P < 0.0001; RESPECTIVELY). FURTHERMORE, IN HCC PATIENTS, THE FREQUENCY OF CDO1 PROMOTER METHYLATION WAS HIGHER IN ADVANCED STAGES (III-IV) (53%) THAN THE EARLY STAGES (I-II) (20%) (P = 0.001). EVALUATION OF THE CDO1 PROMOTER METHYLATION STATUS IN SERUM, IN COMBINATION WITH AFP (> 20 NG/ML), SIGNIFICANTLY IMPROVED THE DIAGNOSTIC VALUE, WITH SENSITIVITY AND SPECIFICITY OF 82.9% AND 75.4%, RESPECTIVELY IN DISTINGUISHING HCC FROM LC AND CHB. IN CONCLUSION, METHYLATION STATUS OF SERUM CDO1 GENE PROMOTER MAY BE HELPFUL IN THE DIAGNOSIS OF HCC AND THE ESTIMATION OF THE HCC STAGES.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
18  3646  37 INCREASED PROTEIN EXPRESSION OF DNA METHYLTRANSFERASE (DNMT) 1 IS SIGNIFICANTLY CORRELATED WITH THE MALIGNANT POTENTIAL AND POOR PROGNOSIS OF HUMAN HEPATOCELLULAR CARCINOMAS. ALTERATION OF DNA METHYLATION IS ONE OF THE MOST CONSISTENT EPIGENETIC CHANGES IN HUMAN CANCERS. DNA METHYLTRANSFERASE (DNMT) 1 IS A MAJOR ENZYME INVOLVED IN ESTABLISHING GENOMIC METHYLATION PATTERNS. MOST OF THE STUDIES CONCERNING DNMT1 EXPRESSION IN HUMAN CANCERS HAVE BEEN PERFORMED ONLY AT THE MRNA LEVEL. TO DIRECTLY EXAMINE DNMT1 PROTEIN EXPRESSION LEVELS DURING HUMAN HEPATOCARCINOGENESIS, 16 HISTOLOGICALLY NORMAL LIVER TISSUES, 51 NONCANCEROUS LIVER TISSUES EXHIBITING CHRONIC HEPATITIS OR CIRRHOSIS, WHICH ARE CONSIDERED TO BE PRECANCEROUS CONDITIONS, AND 53 HEPATOCELLULAR CARCINOMAS (HCCS) WERE SUBJECTED TO IMMUNOHISTOCHEMIC EXAMINATION. IF MORE THAN 20% OF THE CELLS EXHIBITED NUCLEAR DNMT1 STAINING, THE TISSUE SAMPLE WAS CONSIDERED TO BE DNMT1-POSITIVE. DNMT1 IMMUNOREACTIVITY WAS OBSERVED IN 23 (43%) OF THE HCCS, BUT IN NONE (0%) OF THE HISTOLOGICALLY NORMAL LIVER OR NONCANCEROUS LIVER TISSUES EXHIBITING CHRONIC HEPATITIS OR CIRRHOSIS. THE INCIDENCE OF INCREASED DNMT1 PROTEIN EXPRESSION IN HCCS CORRELATED SIGNIFICANTLY WITH POOR TUMOR DIFFERENTIATION (P = 0.0006) AND PORTAL VEIN INVOLVEMENT (P = 0.0002). MOREOVER, THE RECURRENCE-FREE (P = 0.0001) AND OVERALL (P < 0.0001) SURVIVAL RATES OF PATIENTS WITH HCCS EXHIBITING INCREASED DNMT1 PROTEIN EXPRESSION WERE SIGNIFICANTLY LOWER THAN THOSE OF PATIENTS WITH HCCS THAT DID NOT EXHIBIT INCREASED EXPRESSION. INCREASED DNMT1 PROTEIN EXPRESSION MAY PLAY A CRITICAL ROLE IN THE MALIGNANT PROGRESSION OF HCCS AND BE A BIOLOGIC PREDICTOR OF BOTH HCC RECURRENCE AND A POOR PROGNOSIS IN HCC PATIENTS.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
19  5717  38 SIRTUIN-6-DEPENDENT GENETIC AND EPIGENETIC ALTERATIONS ARE ASSOCIATED WITH POOR CLINICAL OUTCOME IN HEPATOCELLULAR CARCINOMA PATIENTS. SIRTUIN 6 (SIRT6) IS A MEMBER OF THE SIRTUIN FAMILY OF NAD+-DEPENDENT DEACETYLASES. GENETIC DELETION OF SIRT6 IN MICE RESULTS IN A SEVERE DEGENERATIVE PHENOTYPE WITH IMPAIRED LIVER FUNCTION AND PREMATURE DEATH. THE ROLE OF SIRT6 IN DEVELOPMENT AND PROGRESSION OF HEPATOCELLULAR CARCINOMA IS CURRENTLY UNKNOWN. WE FIRST INVESTIGATED SIRT6 EXPRESSION IN 153 PRIMARY HUMAN LIVER CANCERS AND IN NORMAL AND CIRRHOTIC LIVERS USING MICROARRAY ANALYSIS. SIRT6 WAS SIGNIFICANTLY DOWN-REGULATED IN BOTH CIRRHOTIC LIVERS AND CANCER. A SIRT6 KNOCKOUT (KO) GENE EXPRESSION SIGNATURE WAS GENERATED FROM PRIMARY HEPATOCTYES ISOLATED FROM 3-WEEK-OLD SIRT6-DEFICIENT ANIMALS. SIRT6-DEFICIENT HEPATOCYTES SHOWED UP-REGULATION OF ESTABLISHED HEPATOCELLULAR CARCINOMA (HCC) BIOMARKERS ALPHA-FETOPROTEIN (AFP), INSULIN-LIKE GROWTH FACTOR 2 (IGF2), H19, AND GLYPICAN-3. FURTHERMORE, DECREASED SIRT6 EXPRESSION WAS OBSERVED IN HEPATOMA CELL LINES THAT ARE KNOWN TO BE APOPTOSIS-INSENSITIVE. RE-EXPRESSION OF SIRT6 IN HEPG2 CELLS INCREASED APOPTOSIS SENSITIVITY TO CD95-STIMULATION OR CHEMOTHERAPY TREATMENT. LOSS OF SIRT6 WAS CHARACTERIZED BY ONCOGENIC CHANGES, SUCH AS GLOBAL HYPOMETHYLATION, AS WELL AS METABOLIC CHANGES, SUCH AS HYPOGLYCEMIA AND INCREASED FAT DEPOSITION. THE HEPATOCYTE-SPECIFIC SIRT6-KO SIGNATURE HAD A PROGNOSTIC IMPACT AND WAS ENRICHED IN PATIENTS WITH POORLY DIFFERENTIATED TUMORS WITH HIGH AFP LEVELS AS WELL AS RECURRENT DISEASE. FINALLY, WE DEMONSTRATED THAT THE SIRT6-KO SIGNATURE POSSESSED A PREDICTIVE VALUE FOR TUMORS OTHER THAN HCC (E.G., BREAST AND LUNG CANCER). CONCLUSION: LOSS OF SIRT6 INDUCES EPIGENETIC CHANGES THAT MAY BE RELEVANT TO CHRONIC LIVER DISEASE AND HCC DEVELOPMENT. DOWN-REGULATION OF SIRT6 AND GENES DYSREGULATED BY LOSS OF SIRT6 POSSESS ONCOGENIC EFFECTS IN HEPATOCARCINOGENESIS. OUR DATA DEMONSTRATE THAT DEFICIENCY IN ONE EPIGENETIC REGULATOR PREDISPOSES A TUMORIGENIC PHENOTYPE THAT ULTIMATELY HAS RELEVANCE FOR OUTCOME OF HCC AND OTHER CANCER PATIENTS.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                     
20  1700  39 DYNAMIC EXPRESSION OF ZNF382 AND ITS TUMOR-SUPPRESSOR ROLE IN HEPATITIS B VIRUS-RELATED HEPATOCELLULAR CARCINOGENESIS. HEPATITIS B VIRUS (HBV) INFECTION IS THE PRIMARY CAUSE OF HEPATOCELLULAR CARCINOMA (HCC). ZINC-FINGER PROTEIN 382 (ZNF382), WHICH BELONGS TO ZINC-FINGER PROTEIN FAMILY, HAS BEEN DOCUMENTED TO BE DOWNREGULATED IN CERTAIN TYPES OF CANCER. HOWEVER, ITS ROLE IN HCC REMAINS LARGELY UNKNOWN. IN THIS STUDY, WE DEMONSTRATED THAT ZNF382 EXPRESSION WAS SIGNIFICANTLY ELEVATED IN HBV-INFECTED LIVER CIRRHOSIS TISSUES RELATIVE TO HBV-NEGATIVE NORMAL LIVER TISSUES AT PROTEIN LEVELS, BUT NOT AT MRNA LEVELS, AND WAS POSITIVELY CORRELATED WITH THE LEVELS OF HBV DNA AND HEPATITIS B VIRUS X PROTEIN (HBX). FURTHER STUDIES REVEALED THAT ZNF382 WAS A TARGET OF MIR-6867, AND HBX PROMOTED THE TRANSLATION OF ZNF382 DURING HBV CHRONIC INFECTION THROUGH ERK-MEDIATED MIR-6867 INHIBITION. IN ADDITION, OUR DATA SHOWED THAT ZNF382 WAS FREQUENTLY DOWNREGULATED BY PROMOTER METHYLATION IN HBV-RELATED HCCS RELATIVE TO HBV-INFECTED LIVER CIRRHOSIS TISSUES, AND DECREASED EXPRESSION OF ZNF382 WAS STRONGLY CORRELATED WITH POOR SURVIVAL IN EARLY-STAGE HCC PATIENTS. FUNCTIONAL STUDIES DEMONSTRATED THAT ZNF382 WAS A POTENT TUMOR SUPPRESSOR IN HCC CELLS THROUGH INHIBITING CELL PROLIFERATION, COLONY FORMATION, MIGRATION, INVASION, AND TUMORIGENIC POTENTIAL IN NUDE MICE, AND INDUCING CELL APOPTOSIS. MECHANISTICALLY, ZNF382 EXERTED ITS TUMOR-SUPPRESSOR FUNCTIONS IN HCC THROUGH TRANSCRIPTIONALLY REPRESSING ITS DOWNSTREAM TARGETS SUCH AS FOS PROTO-ONCOGENE (FOS), JUN PROTO-ONCOGENE (JUN), DISHEVELED SEGMENT POLARITY PROTEIN 2 (DVL2), AND FRIZZLED CLASS RECEPTOR 1 (FZD1), THEREBY IMPAIRING THE ACTIVITIES OF ACTIVATING PROTEIN 1 (AP-1) AND WNT/BETA-CATENIN PATHWAYS AND ACTIVATING P53 SIGNALING. ALTOGETHER, OUR DATA SHOW THAT ZNF382 ACTS AS A TUMOR SUPPRESSOR, AND IS CO-REGULATED BY HBX AND EPIGENETIC MECHANISM IN HBV-RELATED HEPATOCELLULAR CARCINOGENESIS.	2019