1  2436 130 EPIGENETIC SILENCING OF SOCS3 IDENTIFIES A SUBSET OF PROSTATE CANCER WITH AN AGGRESSIVE BEHAVIOR. BACKGROUND: CHRONIC INFLAMMATION AND SUBSEQUENT TISSUTAL ALTERATIONS MAY PLAY A KEY ROLE IN PROSTATE CARCINOGENESIS. IN THIS WAY, MOLECULAR ALTERATIONS OF THE SUPPRESSOR OF CYTOKINE SIGNALING 3 (SOCS3), ONE OF THE MOST IMPORTANT INHIBITORY MOLECULE OF INFLAMMATORY SIGNAL TRANSDUCTION CIRCUITRIES, COULD CONTRIBUTE TO EXPLAIN THE PLEIOTROPIC ROLE OF INTERLEUKIN-6 (IL-6) IN THIS TYPE OF CANCER. METHODS: WE ANALYZED THE METHYLATION STATUS AND MRNA EXPRESSION OF SOCS3 IN 20 BENIGN PROSTATE HYPERPLASIAS (BPH) AND IN 51 PROSTATE CANCER SPECIMENS. WE ANALYZED THE SOCS3 METHYLATION STATUS USING METHYLATION-SPECIFIC PCR. HYPERMETHYLATION WAS CONFIRMED BY SEQUENCING AFTER SUBCLONING. EPIGENETIC SILENCING OF THIS GENE WAS ALSO DEMONSTRATED BY REAL-TIME PCR AND BY IMMUNOHISTOCHEMISTRY. RESULTS AND CORRELATION WITH CLINICAL DATA WERE STATISTICALLY ANALYZED. RESULTS: WE FOUND THAT THE PROMOTER OF SOCS3 WAS METHYLATED IN 39.2% OF PROSTATE CANCER. ON THE CONTRARY, ALL BPH AND NORMAL CONTROLS HAD AN UNMETHYLATED PATTERN. REAL-TIME ANALYSIS SHOWED THAT IN METHYLATED CASES SOCS3 MRNA EXPRESSION WAS REDUCED BY THREE AND FOUR FOLDS AS COMPARED TO BPH AND UNMETHYLATED CASES, RESPECTIVELY. INTERESTINGLY, SOCS3 MRNA LEVEL WAS HIGHER IN UNMETHYLATED PROSTATE CANCER THAN IN BPH. THE IMMUNOHISTOCHEMICAL STAINING ANALYSIS FOR SOCS 3 CONFIRMED MRNA RESULTS. MOREOVER, METHYLATION OF SOCS3 PROMOTER SIGNIFICANTLY ASSOCIATED WITH INTERMEDIATE-HIGH GRADE GLEASON SCORE (P = 0.0007) AND WITH AN UNFAVORABLE CLINICAL OUTCOME (P = 0.0019). CONCLUSIONS: OUR DATA SUGGEST THAT SOCS3 HYPERMETHYLATION MAY BE INVOLVED IN THE PATHOGENESIS OF PROSTATE CANCER AND COULD IDENTIFY A TUMOR SUBSET WITH AN AGGRESSIVE BEHAVIOR.	2011	
                                                                                                                                                                                                                                                                                                                                                                                         
2  5758  36 SOCS3 DELETION IN T LYMPHOCYTES SUPPRESSES DEVELOPMENT OF CHRONIC OCULAR INFLAMMATION VIA UPREGULATION OF CTLA-4 AND EXPANSION OF REGULATORY T CELLS. SUPPRESSORS OF CYTOKINE SIGNALING (SOCS) PROTEINS ARE NEGATIVE-FEEDBACK REGULATORS OF THE JAK/STAT PATHWAY, AND SOCS3 CONTRIBUTES TO HOST IMMUNITY BY REGULATING THE INTENSITY AND DURATION OF CYTOKINE SIGNALS AND INFLAMMATORY RESPONSES. MICE WITH SOCS3 DELETION IN MYELOID CELLS EXHIBIT ENHANCED STAT3 SIGNALING, EXPANSION OF TH1 AND TH17 CELLS, AND DEVELOP SEVERE EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS. INTERESTINGLY, DEVELOPMENT OF THE UNIQUE IL-17/IFN-GAMMA DOUBLE-PRODUCING (TH17/IFN-GAMMA AND TC17/IFN-GAMMA) SUBSETS THAT EXHIBIT STRONG CYTOTOXIC ACTIVITIES AND ARE ASSOCIATED WITH PATHOGENESIS OF SEVERAL AUTOIMMUNE DISEASES HAS RECENTLY BEEN SHOWN TO DEPEND ON EPIGENETIC SUPPRESSION OF SOCS3 EXPRESSION, FURTHER SUGGESTING INVOLVEMENT OF SOCS3 IN AUTOIMMUNITY AND TUMOR IMMUNITY. IN THIS STUDY, WE GENERATED MICE WITH SOCS3 DELETION IN THE CD4 T CELL COMPARTMENT (CD4-SOCS3 KNOCKOUT [KO]) TO DETERMINE IN VIVO EFFECTS OF THE LOSS OF SOCS3 IN THE T CELL-MEDIATED AUTOIMMUNE DISEASE, EXPERIMENTAL AUTOIMMUNE UVEITIS (EAU). IN CONTRAST TO THE EXACERBATION OF EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS IN MYELOID-SPECIFIC SOCS3-DELETED MICE, CD4-SOCS3KO MICE WERE PROTECTED FROM ACUTE AND CHRONIC UVEITIS. PROTECTION FROM EAU CORRELATED WITH ENHANCED EXPRESSION OF CTLA-4 AND EXPANSION OF IL-10-PRODUCING REGULATORY T CELLS WITH AUGMENTED SUPPRESSIVE ACTIVITIES. WE FURTHER SHOW THAT SOCS3 INTERACTS WITH CTLA-4 AND NEGATIVELY REGULATES CTLA-4 LEVELS IN T CELLS, PROVIDING A MECHANISTIC EXPLANATION FOR THE EXPANSION OF REGULATORY T CELLS IN CD4-SOCS3 DURING EAU. CONTRARY TO IN VITRO EPIGENETIC STUDIES, TH17/IFN-GAMMA AND TC17/IFN-GAMMA POPULATIONS WERE MARKEDLY REDUCED IN CD4-SOCS3KO, SUGGESTING THAT SOCS3 PROMOTES EXPANSION OF THE TH17/IFN-GAMMA SUBSET ASSOCIATED WITH DEVELOPMENT OF SEVERE UVEITIS. THUS, SOCS3 IS A POTENTIAL THERAPEUTIC TARGET IN UVEITIS AND OTHER AUTOINFLAMMATORY DISEASES.	2013	
                                                                                                                        
3   388  43 AN INTEGRATED ANALYSIS OF SOCS1 DOWN-REGULATION IN HBV INFECTION-RELATED HEPATOCELLULAR CARCINOMA. PERSISTENT INFLAMMATION TOGETHER WITH GENETIC/EPIGENETIC ABERRATIONS IS STRONGLY ASSOCIATED WITH CHRONIC HEPATITIS B VIRUS (HBV) INFECTION-RELATED HEPATOCARCINOGENESIS. HERE, WE INVESTIGATED THE ALTERATIONS OF THE SUPPRESSOR OF CYTOKINE SIGNALLING (SOCS) FAMILY GENES IN HBV-RELATED HEPATOCELLULAR CARCINOMA (HCC). A TOTAL OF 116 PATIENTS WITH HCC WERE ENROLLED IN THIS STUDY. THE METHYLATION STATUSES OF SOCS1-7 AND CISH GENES WERE QUANTITATIVELY MEASURED AND CLINICOPATHOLOGICAL SIGNIFICANCE OF SOCS1 METHYLATION WAS STATISTICALLY ANALYSED. THE GENE COPY NUMBER VARIATION WAS ASSAYED BY ACGH. LUCIFERASE REPORTER ASSAY AND WESTERN BLOT WERE USED TO DETECT THE INVOLVEMENT OF SOCS1 IN P53 SIGNALLING. WE FOUND HIGH FREQUENCIES OF SOCS1 GENE HYPERMETHYLATION IN BOTH TUMOUR (56.03%) AND ADJACENT NONTUMOUR TISSUES (54.31%), BUT TUMOUR TISSUES EXHIBITED INCREASED METHYLATION INTENSITY (24.01% VS 13.11%, P < 0.0001), PARTICULARLY IN PATIENTS WITH LARGER TUMOUR SIZE OR CIRRHOSIS BACKGROUND (P < 0.0001). IN ADDITION, THE FREQUENCY AND INTENSITY OF SOCS1 HYPERMETHYLATION IN TUMOUR TISSUES WERE BOTH SIGNIFICANTLY HIGHER THAN THOSE IN NONTUMOUR TISSUES IN MALE GENDER PATIENTS AND IN PATIENTS >/=45 YEARS OLD (P = 0.0214 AND P < 0.0001, P = 0.0232 AND P < 0.0001, RESPECTIVELY). SOCS1 GENE DELETION WAS FOUND IN 8 OF 25 ACGH ASSAYED TUMOUR SPECIMENS, WHICH WAS ASSOCIATED WITH LOWER SOCS1 MRNA EXPRESSION (P = 0.0448). FURTHERMORE, ECTOPIC SOCS1 OVEREXPRESSION COULD ACTIVATE THE P53 SIGNALLING PATHWAY IN HCC CELL LINES. HYPERMETHYLATION OF SOCS2-7 AND CISH GENES WAS SELDOM FOUND IN HCC. OUR RESULTS SUGGESTED THAT THE GENE LOSS AND EPIGENETIC SILENCING OF SOCS1 WERE STRONGLY ASSOCIATED WITH HBV-RELATED HCC.	2014	
                                                                                                                                                                                                                                                                                                                                                                               
4  3276  27 HEPATOCYTE GROWTH CONTROL BY SOCS1 AND SOCS3. THE EXTRAORDINARY CAPACITY OF THE LIVER TO REGENERATE FOLLOWING INJURY IS DEPENDENT ON COORDINATED AND REGULATED ACTIONS OF CYTOKINES AND GROWTH FACTORS. WHEREAS HEPATOCYTE GROWTH FACTOR (HGF) AND EPIDERMAL GROWTH FACTOR (EGF) ARE DIRECT MITOGENS TO HEPATOCYTES, INFLAMMATORY CYTOKINES SUCH AS TNFALPHA AND IL-6 ALSO PLAY ESSENTIAL ROLES IN THE LIVER REGENERATION PROCESS. THESE CYTOKINES AND GROWTH FACTORS ACTIVATE DIFFERENT SIGNALING PATHWAYS IN A SEQUENTIAL MANNER TO ELICIT HEPATOCYTE PROLIFERATION. THE KINETICS AND MAGNITUDE OF THESE HEPATOCYTE-ACTIVATING STIMULI ARE TIGHTLY REGULATED TO ENSURE RESTORATION OF A FUNCTIONAL LIVER MASS WITHOUT CAUSING UNCONTROLLED CELL PROLIFERATION. HEPATOCYTE PROLIFERATION CAN BECOME DEREGULATED UNDER CONDITIONS OF CHRONIC INFLAMMATION, LEADING TO ACCUMULATION OF GENETIC ABERRATIONS AND EVENTUAL NEOPLASTIC TRANSFORMATION. AMONG THE CONTROL MECHANISMS THAT REGULATE HEPATOCYTE PROLIFERATION, NEGATIVE FEEDBACK INHIBITION BY THE 'SUPPRESSOR OF CYTOKINE SIGNALING (SOCS)' FAMILY PROTEINS SOCS1 AND SOCS3 PLAY CRUCIAL ROLES IN ATTENUATING CYTOKINE AND GROWTH FACTOR SIGNALING. LOSS OF SOCS1 OR SOCS3 IN THE MOUSE LIVER INCREASES THE RATE OF LIVER REGENERATION AND RENDERS HEPATOCYTES SUSCEPTIBLE TO NEOPLASTIC TRANSFORMATION. THE FREQUENT EPIGENETIC REPRESSION OF THE SOCS1 AND SOCS3 GENES IN HEPATOCELLULAR CARCINOMA HAS STIMULATED RESEARCH IN UNDERSTANDING THE GROWTH REGULATORY MECHANISMS OF SOCS1 AND SOCS3 IN HEPATOCYTES. WHEREAS SOCS3 IS IMPLICATED IN REGULATING JAK-STAT SIGNALING INDUCED BY IL-6 AND ATTENUATING EGFR SIGNALING, SOCS1 IS CRUCIAL FOR THE REGULATION OF HGF SIGNALING. THESE TWO PROTEINS ALSO MODULE THE FUNCTIONS OF CERTAIN KEY PROTEINS THAT CONTROL THE CELL CYCLE. IN THIS REVIEW, WE DISCUSS THE CURRENT UNDERSTANDING OF THE FUNCTIONS OF SOCS1 AND SOCS3 IN CONTROLLING HEPATOCYTE PROLIFERATION, AND ITS IMPLICATIONS TO LIVER HEALTH AND DISEASE.	2019	
                                                                                                                                                                                                                             
5  2842  42 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	
                                                                                                                                                                                                                                                      
6  1968  42 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
7  2129  30 EPIGENETIC INACTIVATION OF SUPPRESSORS OF CYTOKINE SIGNALLING IN PHILADELPHIA-NEGATIVE CHRONIC MYELOPROLIFERATIVE DISORDERS. PH-NEGATIVE CHRONIC MYELOPROLIFERATIVE DISORDERS (CMPD) ARE CHARACTERIZED BY CONSTITUTIVE JANUS KINASE-SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION (JAK-STAT) ACTIVATION. SOCS3, SOCS1 AND PTPN6 (SHP1) ARE NEGATIVE REGULATORS OF THE JAK-STAT PATHWAY. WE INVESTIGATED EPIGENETIC AND GENETIC INACTIVATION OF SOCS3, SOCS1 AND PTPN6 IN 112 CMPD AND 20 ACUTE MYELOID LEUKAEMIA (AML) POST-CMPD. SOCS3 METHYLATION OCCURRED AT HIGH FREQUENCY IN BOTH CMPD (46/112; 41.1%) AND AML POST-CMPD (10/17; 58.8%) AND WAS ASSOCIATED WITH TRANSCRIPTIONAL SILENCING. IN CONTRAST, METHYLATION OF SOCS1 AND PTPN6 WAS OBSERVED IN ONLY A FRACTION OF CMPD (15/112, 13.4% FOR SOCS1; AND 8/112, 7.1% FOR PTPN6) AND AML POST-CMPD (3/20, 15% FOR SOCS1; AND 1/20, 5% FOR PTPN6). NO SOMATIC MUTATIONS OF SOCS1 WERE FOUND IN CMPD. SOCS3, SOCS1 AND PTPN6 METHYLATION OCCURRED IN BOTH JAK2V617F-POSITIVE (35.1% FOR SOCS3; 14.9% FOR SOCS1; 8.1% FOR PTPN6) AND JAK2V617F-NEGATIVE (57.1% FOR SOCS3; 14.3% FOR SOCS1; AND 9.5% FOR PTPN6) CMPD. THESE DATA INDICATE THAT METHYLATION OF SOCS3 AND, TO A LESSER EXTENT, SOCS1 AND PTPN6 IS A FREQUENT EVENT IN BOTH JAK2V617F-POSITIVE AND -NEGATIVE CMPD AND MAY ACT AS AN ALTERNATIVE OR COMPLEMENTARY MECHANISM TO JAK2 MUTATIONS, ENHANCING CYTOKINE SIGNAL TRANSDUCTION. THE FREQUENT INACTIVATION OF SOCS3 IS A NOVEL FINDING IN CMPD WITH POTENTIAL IMPLICATIONS FOR THE MOLECULAR PATHOLOGY OF THESE DISORDERS.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
8  4231  45 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	
                                                                                                                     
9  4601  45 NDRG2 MRNA LEVELS AND MIR-28-5P AND MIR-650 ACTIVITY IN CHRONIC LYMPHOCYTIC LEUKEMIA. BACKGROUND: NDRG2 IS IDENTIFIED AS A TUMOR SUPPRESSOR GENE IN MANY TUMORS, AND FUNCTIONS IN CELL PROLIFERATION, DIFFERENTIATION AND APOPTOSIS. RECENT DATA INDICATE THAT NDRG2 EXPRESSION IS UP-REGULATED BY TP53. MOREOVER, PROPOSED MECHANISMS OF NDRG2 INACTIVATION INCLUDE EPIGENETIC SILENCING OF THE NDRG2 PROMOTER AND DOWN-REGULATION BY MICRORNAS (MIRNAS). HOWEVER, FEW STUDIES HAVE EVER BEEN DONE ON THE ROLE OF NDRG2 AND THE NDRG2-REGULATING MIRNAS INTERFERENCE IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). METHODS: NDRG2 AND MICRORNAS MRNA LEVELS IN CLL SUBJECTS WERE ASSESSED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QRT-PCR). THE DUAL-LUCIFERASE REPORTER ASSAY WAS PERFORMED TO DETERMINE NDRG2-RELATED MIRNAS. LOW EXPRESSION OF MATURE EXOGENOUS MIRNAS IN CLL CELLS WAS ESTABLISHED BY TRANSIENT TRANSFECTION. NDRG2 PROTEIN LEVELS IN CLL CELLS WERE DETECTED BY WESTERN BLOT. IN ADDITION, FLOW CYTOMETRY WAS CONDUCTED TO EXAMINE THE APOPTOSIS OF CLL CELLS. RESULTS: LOWER EXPRESSION OF NDRG2 WAS FOUND IN THE B-CELLS FROM 102 CLL PATIENTS COMPARED THE 40 NORMAL SUBJECTS (P < 0.001). PATIENTS WITH ADVANCED BINET STAGE (P = 0.001), HIGH LACTATE DEHYDROGENASE (LDH) LEVEL (P = 0.036), UN-MUTATED IMMUNOGLOBULIN HEAVY CHAIN VARIABLE REGION GENE (IGHV) (P = 0.004) AND THOSE WITH P53 ABERRATIONS (P < 0.001) HAD A MARKEDLY LOWER LEVELS OF NDRG2 MRNA. THIS DECREASE WAS ASSOCIATED WITH BRIEFER TIME-TO-TREATMENT (P = 0.001) AND POORER SURVIVAL (P < 0.001). HIGH EXPRESSION OF MIR-28-5P AND MIR-650 WAS ASSOCIATED WITH BINET B/C STAGE (P = 0.044) AND IGHV UN-MUTATED (P = 0.011), AS WELL AS BINET B/C STAGE (P = 0.013) AND P53 ABERRATIONS (P = 0.037), RESPECTIVELY. INHIBITION OF MIR-28-5P OR MIR-650 COULD INDUCE MORE APOPTOSIS IN CLL CELLS WITH GERMLINE TP53. CONCLUSIONS: NDRG2 MRNA LEVELS MIGHT BE A USEFUL PROGNOSTIC VARIABLE FOR PATIENTS OF CLL AND UP-REGULATING NDRG2 TRANSCRIPTION MAY BE A THERAPY APPROACH IN CLL WITHOUT P53 ABERRATIONS.	2018	
                                                                                                                                             
10  4903  31 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
11  2844  30 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
12  6770  42 [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	

13   703  37 BTLA EXPRESSION IN CLL: EPIGENETIC REGULATION AND IMPACT ON CLL B CELL PROLIFERATION AND ABILITY TO IL-4 PRODUCTION. IN OUR PREVIOUS STUDY, WHILE CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) CASES SHOWED HIGHER LEVELS OF B AND T LYMPHOCYTE ATTENUATOR (BTLA) MRNA COMPARED TO CONTROLS, LOWER BTLA PROTEIN EXPRESSION WAS OBSERVED IN CASES COMPARED TO CONTROLS. HENCE WE HYPOTHESIZE THAT MICRO RNA (MIR) 155-5P REGULATES BTLA EXPRESSION IN CLL. IN LINE WITH EARLIER DATA, EXPRESSION OF BTLA MRNA AND MIR-155-5P IS ELEVATED IN CLL (P = 0.034 AND P = 0.0006, RESPECTIVELY) AS WELL AS IN MEC-1 CELL LINE (P = 0.009 AND 0.016, RESPECTIVELY). INHIBITION OF MIR-155-5P PARTIALLY RESTORED BTLA PROTEIN EXPRESSION IN CLL PATIENTS (P = 0.01) AND IN MEC-1 CELL LINES (P = 0.058). ADDITIONALLY, WE AIMED TO EVALUATE THE SIGNIFICANCE OF BTLA DEFICIENCY IN CLL CELLS ON PROLIFERATION AND IL-4 PRODUCTION OF B CELLS. WE FOUND THAT SECRETION OF IL-4 IS NOT DEPENDENT ON BTLA EXPRESSION, SINCE FRACTIONS OF BTLA POSITIVE AND BTLA NEGATIVE B CELLS EXPRESSING INTRACELLULAR IL-4 WERE SIMILAR IN CLL PATIENTS AND CONTROLS. WE DEMONSTRATED THAT IN CONTROLS THE FRACTION OF PROLIFERATING CELLS IS LOWER IN BTLA POSITIVE THAN IN BTLA NEGATIVE B CELLS (P = 0.059), WHICH WAS NOT OBSERVED IN CLL. HOWEVER, THE FREQUENCY OF BTLA POSITIVE KI67+ B CELLS IN CLL WAS HIGHER COMPARED TO CORRESPONDING CELLS FROM CONTROLS (P = 0.055) WHILE THERE WERE NO DIFFERENCES BETWEEN THE EXAMINED GROUPS REGARDING FREQUENCY OF BTLA NEGATIVE KI67+ B CELLS. OUR STUDIES SUGGEST THAT MIR-155-5P IS INVOLVED IN BTLA DEFICIENCY, AFFECTING PROLIFERATION OF CLL B CELLS, WHICH MAY BE ONE OF THE MECHANISMS RESPONSIBLE FOR CLL PATHOGENESIS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
14  3825  38 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	
                                                                                                                                                                                                                                                                                                                                                                                      
15  4248  46 METHYLATION STATUS, MRNA AND PROTEIN EXPRESSION OF THE SMAD4 GENE IN PATIENTS WITH NON-MELANOCYTIC SKIN CANCERS. BACKGROUND: SMAD4 IS A POTENT TUMOR SUPPRESSOR. SMAD4 LOSS INCREASES GENOMIC INSTABILITY AND PLAYS A CRITICAL ROLE IN THE DNA DAMAGE RESPONSE THAT LEADS TO SKIN CANCER DEVELOPMENT. WE AIMED TO INVESTIGATE SMAD4 METHYLATION EFFECTS ON MRNA AND PROTEIN EXPRESSION OF SMAD4 IN CANCER AND HEALTHY TISSUES FROM PATIENTS WITH BASAL CELL CARCINOMA (BCC), CUTANEOUS SQUAMOUS CELL CARCINOMA (CSCC), AND BASOSQUAMOUS SKIN CANCER (BSC). METHODS AND RESULTS: THE STUDY INCLUDED 17 BCC, 24 CSCC AND NINE BSC PATIENTS. DNA AND RNA WERE ISOLATED FROM CANCEROUS AND HEALTHY TISSUES FOLLOWING PUNCH BIOPSY. METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (PCR) AND REAL-TIME QUANTITATIVE PCR METHODS WERE USED TO EXAMINE SMAD4 PROMOTER METHYLATION AND SMAD4 MRNA LEVELS, RESPECTIVELY. THE PERCENTAGE AND INTENSITY OF STAINING OF THE SMAD4 PROTEIN WERE DETERMINED BY IMMUNOHISTOCHEMISTRY. THE PERCENTAGE OF SMAD4 METHYLATION WAS INCREASED IN THE PATIENTS WITH BCC (P = 0.007), CSCC (P = 0.004), AND BSC (P = 0.018) COMPARED TO THE HEALTHY TISSUE. SMAD4 MRNA EXPRESSION WAS DECREASED IN THE PATIENTS WITH BCC (P<0.001), CSCC (P<0.001), AND BSC (P = 0.008). THE STAINING CHARACTERISTIC OF SMAD4 PROTEIN WAS NEGATIVE IN THE CANCER TISSUES OF THE PATIENTS WITH CSCC (P = 0.00). LOWER SMAD4 MRNA LEVELS WERE OBSERVED IN THE POORLY DIFFERENTIATED CSCC PATIENTS (P = 0.001). THE STAINING CHARACTERISTICS OF THE SMAD4 PROTEIN WERE RELATED TO AGE AND CHRONIC SUN EXPOSURE. CONCLUSIONS: HYPERMETHYLATION OF SMAD4 AND REDUCED SMAD4 MRNA EXPRESSION WERE FOUND TO PLAY A ROLE IN THE PATHOGENESIS OF BCC, CSCC, AND BSC. A DECREASE IN SMAD4 PROTEIN EXPRESSION LEVEL WAS OBSERVED ONLY IN CSCC PATIENTS. THIS SUGGESTS THAT EPIGENETIC ALTERATIONS TO THE SMAD4 GENE ARE ASSOCIATED WITH CSCC. TRIAL REGISTRATION: THE NAME OF THE TRIAL REGISTER: SMAD4 METHYLATION AND EXPRESSION LEVELS IN NON-MELANOCYTIC SKIN CANCERS; SMAD4 PROTEIN POSITIVITY. THE REGISTRATION NUMBER: NCT04759261 ( HTTPS://CLINICALTRIALS.GOV/CT2/RESULTS?TERM=NCT04759261 ).	2023	
                                                            
16  2440  32 EPIGENETIC SILENCING OF TUMOR SUPPRESSOR LONG NON-CODING RNA BM742401 IN CHRONIC LYMPHOCYTIC LEUKEMIA. BM742401 IS A TUMOR SUPPRESSOR LNCRNA DOWNREGULATED IN GASTRIC CANCER. AS THE PROMOTER REGION AND THE ENTIRE TRANSCRIPT ARE EMBEDDED IN A CPG ISLAND, WE POSTULATED THAT BM742401 IS A TUMOR SUPPRESSOR LNCRNA INACTIVATED BY DNA METHYLATION IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). THE PROMOTER OF BM742401 WAS UNMETHYLATED IN NORMAL CONTROLS INCLUDING THREE EACH OF NORMAL BONE MARROW, PERIPHERAL BLOOD BUFFY COATS, AND CD19-SORTED PERIPHERAL B-CELLS, BUT METHYLATED IN FOUR (57.1%) CLL CELL LINES. METHYLATION OF BM742401 CORRELATED INVERSELY WITH EXPRESSION. IN THE COMPLETELY METHYLATED WAC3CD5+ CLL CELLS, 5-AZA-2'-DEOXYCYTIDINE TREATMENT LED TO PROMOTER DEMETHYLATION AND RE-EXPRESSION OF BM742401 TRANSCRIPT. FUNCTIONALLY, STABLE OVEREXPRESSION OF BM742401 RESULTED IN INHIBITION OF CELLULAR PROLIFERATION AND ENHANCED APOPTOSIS THROUGH CASPASE-9-DEPENDENT INTRINSIC BUT NOT CASPASE-8-DEPENDENT EXTRINSIC APOPTOSIS PATHWAY, SUGGESTING A TUMOR SUPPRESSOR ROLE OF BM742401 IN CLL. IN PRIMARY CLL SAMPLES, METHYLATION OF BM742401 WAS DETECTED IN 43/98 (43.9%) OF PATIENTS. MOREOVER, AMONG CLL PATIENTS WITH STANDARD-RISK CYTOGENETIC ABERRATIONS, METHYLATION OF BM742401 CORRELATED WITH ADVANCED RAI STAGE (>/= STAGE 2)(P = 0.002). FURTHERMORE, BM742401 METHYLATION WAS ASSOCIATED WITH MIR-129-2 METHYLATION (P = 0.05). BM742401 IS A TUMOR SUPPRESSOR LNCRNA FREQUENTLY METHYLATED IN CLL. THE MECHANISM OF BM742401 AS A TUMOR SUPPRESSOR WARRANTS FURTHER STUDIES.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
17  5795  41 STAT3 INDUCTION OF MIR-146B FORMS A FEEDBACK LOOP TO INHIBIT THE NF-KAPPAB TO IL-6 SIGNALING AXIS AND STAT3-DRIVEN CANCER PHENOTYPES. INTERLEUKIN-6 (IL-6)-MEDIATED ACTIVATION OF SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION 3 (STAT3) IS A MECHANISM BY WHICH CHRONIC INFLAMMATION CAN CONTRIBUTE TO CANCER AND IS A COMMON ONCOGENIC EVENT. WE DISCOVERED A PATHWAY, THE LOSS OF WHICH IS ASSOCIATED WITH PERSISTENT STAT3 ACTIVATION IN HUMAN CANCER. WE FOUND THAT THE GENE ENCODING THE TUMOR SUPPRESSOR MICRORNA MIR-146B IS A DIRECT STAT3 TARGET GENE, AND ITS EXPRESSION WAS INCREASED IN NORMAL BREAST EPITHELIAL CELLS BUT DECREASED IN TUMOR CELLS. METHYLATION OF THE MIR-146B PROMOTER, WHICH INHIBITED STAT3-MEDIATED INDUCTION OF EXPRESSION, WAS INCREASED IN PRIMARY BREAST CANCERS. MOREOVER, WE FOUND THAT MIR-146B INHIBITED NUCLEAR FACTOR KAPPAB (NF-KAPPAB)-DEPENDENT PRODUCTION OF IL-6, SUBSEQUENT STAT3 ACTIVATION, AND IL-6/STAT3-DRIVEN MIGRATION AND INVASION IN BREAST CANCER CELLS, THEREBY ESTABLISHING A NEGATIVE FEEDBACK LOOP. IN ADDITION, HIGHER EXPRESSION OF MIR-146B WAS POSITIVELY CORRELATED WITH PATIENT SURVIVAL IN BREAST CANCER SUBTYPES WITH INCREASED IL6 EXPRESSION AND STAT3 PHOSPHORYLATION. OUR RESULTS IDENTIFY AN EPIGENETIC MECHANISM OF CROSSTALK BETWEEN STAT3 AND NF-KAPPAB RELEVANT TO CONSTITUTIVE STAT3 ACTIVATION IN MALIGNANCY AND THE ROLE OF INFLAMMATION IN ONCOGENESIS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
18  1495  31 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
19  1700  34 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	
                                                                                                                                                                                                                            
20  3294  44 HIGH INCIDENCE OF MGMT AND RARBETA PROMOTER METHYLATION IN PRIMARY GLIOBLASTOMAS: ASSOCIATION WITH HISTOPATHOLOGICAL CHARACTERISTICS, INFLAMMATORY MEDIATORS AND CLINICAL OUTCOME. GLIOBLASTOMAS, THE MOST FREQUENT PRIMARY BRAIN TUMORS IN ADULTS, ARE CHARACTERIZED BY A HIGHLY AGGRESSIVE, INFLAMMATORY AND ANGIOGENIC PHENOTYPE. METHYLATION OF CPG ISLANDS IN CANCER-RELATED GENES MAY SERVE AS AN EPIGENETIC BIOMARKER FOR GLIOBLASTOMA DIAGNOSIS AND PROGNOSIS. THE AIM OF THIS STUDY WAS TO ANALYZE THE METHYLATION STATUS OF FOUR CRITICAL TUMOR-ASSOCIATED GENES (MGMT, RARBETA, RASSF1A, CDH13), AND INVESTIGATE POSSIBLE LINKS WITH INFLAMMATORY (INTERLEUKIN [IL]-6, IL-8) AND ANGIOGENIC MEDIATORS (VASCULAR ENDOTHELIAL GROWTH FACTOR [VEGF], CYCLOOXYGENASE [COX]-2) AND CLINICAL OUTCOME IN 23 GLIOMA SAMPLES (6 GRADE II ASTROCYTOMAS, 17 GRADE IV GLIOBLASTOMAS). RARBETA AND MGMT GENES WERE MORE FREQUENTLY METHYLATED IN 70.58% AND 58.8% OF GLIOBLASTOMAS, RESPECTIVELY. RASSF1A AND CDH13 DISPLAYED A SIMILAR METHYLATION FREQUENCY (23.52%) IN GLIOBLASTOMAS. NO GENE METHYLATION WAS OBSERVED IN GRADE II ASTROCYTOMAS. TUMOR GRADE CORRELATED POSITIVELY WITH MGMT AND RARBETA METHYLATION (P = 0.005 AND P = 0.019, RESPECTIVELY) AND THE EXTENT OF NECROSIS (P = 0.001 AND P = 0.003). INTERESTINGLY, THE MARKER OF CHRONIC INFLAMMATION, IL-6, WAS POSITIVELY ASSOCIATED WITH METHYLATION OF MGMT (P = 0.004), RARBETA (P = 0.002), AND RASSF1A (P = 0.0081) AS WELL AS THE TOTAL NUMBER OF METHYLATED GENES (P < 0.0001), INDICATING THE IMPORTANT ROLE OF IL-6 IN MAINTAINING PROMOTER METHYLATION OF THESE GENES. VEGF EXPRESSION CORRELATED POSITIVELY WITH MGMT AND RARBETA METHYLATION ALTHOUGH THESE RELATIONSHIPS WERE OF MARGINAL SIGNIFICANCE (P = 0.0679 AND P = 0.0757). KAPLAN-MEIER UNIVARIATE SURVIVAL ANALYSIS INDICATED AN UNFAVORABLE SURVIVAL PERIOD IN PATIENTS WITH MGMT METHYLATION COMPARED WITH THOSE WITHOUT METHYLATION (P = 0.0474). OUR STUDY HIGHLIGHTS THE IMPLICATION OF MGMT AND RARBETA METHYLATION IN THE AGGRESSIVE PHENOTYPE OF PRIMARY GLIOBLASTOMAS. THE ASSOCIATION OF MGMT METHYLATION WITH CLINICAL OUTCOME INDICATES ITS POTENTIAL PROGNOSTIC VALUE.	2010