1  2942 106 GENETIC AND EPIGENETIC ALTERATIONS OF LTF AT 3P21.3 IN NASOPHARYNGEAL CARCINOMA. TO INVESTIGATE THE ROLES OF LACTOTRANSFERRIN GENE (LTF, ALSO REFERRED TO AS THE LACTOFERRIN GENE, LF), LOCATED AT 3P21.3 WITHIN THE COMMON MINIMAL DELETION REGION, IN THE PATHOGENESIS OF NASOPHARYNGEAL CARCINOMA (NPC), WE FIRST DETECTED ITS EXPRESSION LEVEL IN 33 PRIMARY NPC TISSUES AND 15 CHRONIC NASOPHARYNGITIS TISSUES. ABSENT EXPRESSION OR DOWNREGULATION OF LTF WERE OBSERVED IN 76% (25 OF 33) OF PRIMARY NPC TISSUES. WE FURTHER FOUND THAT 25% (5 OF 20) OF NPC SPECIMENS HAD LOSS OF HETEROZYGOSITY (LOH) AT THE LTF LOCUS. LTF MUTATION ASSESSED BY POLYMERASE CHAIN REACTION SINGLE-STRAND CONFORMATION POLYMORPHISM (PCR-SSCP) AND DNA SEQUENCING WAS NOTED IN 30% (6 OF 20) OF PRIMARY NPC TISSUES. IN ADDITION, HYPER-METHYLATION OF LTF PROMOTER REGION WAS FOUND IN 63.6% (21 OF 33) OF PRIMARY NPC SAMPLES BUT NOT IN CHRONIC NASOPHARYNGITIS TISSUES. THE LTF TRANSCRIPTS IN NPC CELL LINES INCREASED UPON TREATMENT WITH THE DEMETHYLATION COMPOUND, 5-AZA-2-DEOXYCYTIDINE. IN CONCLUSION, OUR DATA INDICATE THAT TWO-HIT SILENCING OF LTF THROUGH GENETIC AND EPIGENETIC CHANGES MAY BE A COMMON AND IMPORTANT EVENT IN THE CARCINOGENESIS OF NPC.	2006	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
2  1996  49 EPIGENETIC AND GENETIC ALTERATIONS OF THE EDNRB GENE IN NASOPHARYNGEAL CARCINOMA. BACKGROUND: LOSS OF HETEROZYGOSITY (LOH) AT 13Q22 IS A COMMON EVENT IN NASOPHARYNGEAL CARCINOMA (NPC). EDNRB GENE LOCATED AT 13Q22 HAS BEEN DEMONSTRATED TO BE HYPERMETHYLATED IN SOME KINDS OF TUMORS. IN THE CURRENT STUDY, WE FOCUSED ON THE EPIGENETIC AND GENETIC ALTERATIONS OF EDNRB IN NPC. METHODS: THE MRNA EXPRESSION OF EDNRB WAS DETECTED BY SEMIQUANTITATIVE RT-PCR AND REAL-TIME QUANTITATIVE PCR IN 49 NPC AND 12 CHRONIC NASOPHARYNGITIS BIOPSIES. THE METHYLATION AND LOH STATUS OF EDNRB WERE EXAMINED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION, MICROSATELLITE PCR AND SEQUENCING. WE ALSO EXAMINED THE MRNA EXPRESSION OF EDNRB IN FOUR NPC CELL LINES AFTER 5-AZA-2'-DEOXYCYTIDINE TREATMENT. RESULTS: EDNRB WAS DOWNREGULATED IN PRIMARY NPC TISSUES AND NPC CELL LINES, AND A RELATIVELY HIGHER METHYLATION LEVEL OF EDNRB WAS FOUND IN NPC BIOPSIES (84%) COMPARED TO THAT IN CHRONIC NASOPHARYNGITIS BIOPSIES (42%). TREATMENT OF NPC CELL LINES WITH 5-AZA-2'-DEOXYCYTIDINE ACTIVATED EDNRB EXPRESSION. LOH OF EDNRB GENE WAS ALSO FOUND AT TWO MICROSATELLITE SITES WITH RATIOS OF 6.25 AND 16.67% IN NPC. CONCLUSION: OUR RESULTS SUGGESTED THAT EDNRB EXPRESSION MAY BE AFFECTED BY ABERRANT PROMOTER METHYLATION AND GENE DELETION AND MAY PLAY A ROLE IN THE DEVELOPMENT OF NPC.	2007	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
3  6816  51 [EXPRESSION, GENETIC AND EPIGENETIC ALTERATIONS OF LTF GENE IN NASOPHARYNGEAL CARCINOMA CELL LINES]. OBJECTIVE: TO INVESTIGATE THE EXPRESSION OF LTF MRNA IN SEVERAL NASOPHARYNGEAL CANCER (NPC) CELL LINES, AND ANALYZE THE RELATIONSHIP BETWEEN THE GENETIC AND EPIGENETIC CHANGES AND EXPRESSION OF LTF GENE. METHODS: THE EXPRESSION LEVEL OF LTF WAS DETECTED IN NPC CELL LINES HNE1, HNE2, HNE3, CNE1, CNE2, 5-8F, 6-10B CELLS AND TISSUES OF 15 CASES OF CHRONIC NASOPHARYNGITIS BY RT-PCR. THE LTF PROTEIN LEVEL WAS ANALYZED BY WESTERN BLOTTING IN 6-10B CELLS. THEN LOH, MUTATION AND METHYLATION STATUS OF LTF WAS EXAMINED BY MICROSATELLITES ANALYSIS, PCR-SSCP, MSP AND BISULFITE GENOMIC SEQUENCING, RESPECTIVELY. RESULTS: 15 CHRONIC NASOPHARYNGITIS TISSUES SHOWED STABLE LTF EXPRESSION, WHILE THERE WERE WEAK EXPRESSION IN 6-10B CELLS AND ABSENT EXPRESSION IN REMAINING DETECTED NPC CELL LINES. THERE WAS A SIGNIFICANTLY LOWER LTF EXPRESSION IN CHRONIC NASOPHARYNGITIS TISSUES (Z = -3.738, P = 0.000). NO LTF PROTEIN EXPRESSION WAS OBSERVED IN 6-10B CELLS. LOH ANALYSIS DEMONSTRATED THAT ALLELE LOSS OF LTF WASN'T FOUND IN NPC CELL LINES. LTF MUTATION WAS NOTED IN 14.3% (1/7) OF NPC CELL LINES. DNA SEQUENCING CONFIRMED THE MUTATION POINT IN THE PROMOTER REGION (-305 BP TO -50 BP) WAS AT -218 BP (DEL T) OF LTF GENE IN THE HNE1 CELL LINE. METHYLATION OF LTF GENE WAS NOT FOUND IN CHRONIC NASOPHARYNGITIS. HOWEVER, METHYLATION OF LTF PROMOTER WAS DETECTED IN ALL NPC CELL LINES. LTF MRNA EXPRESSION WAS INCREASED IN 5-8F AND 6-10B CELL LINES AFTER TREATMENT WITH 5-AZA-2-DEOXYCYTIDINE. CONCLUSION: THERE IS AN INACTIVATION OF EXPRESSION OF LTF GENE IN THE NPC CELL LINES. ITS MOLECULAR MECHANISM MAY BE RELATED WITH METHYLATION OF PROMOTER REGION AND DELETION MUTATION.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
4  3627  60 INACTIVATION OF LARS2, LOCATED AT THE COMMONLY DELETED REGION 3P21.3, BY BOTH EPIGENETIC AND GENETIC MECHANISMS IN NASOPHARYNGEAL CARCINOMA. ALLELIC LOSS OF CHROMOSOME 3P, INCLUDING THE 3P21.3 REGION, IS FOUND IN 95-100% OF PRIMARY NASOPHARYNGEAL CARCINOMA (NPC) BIOPSIES, SUGGESTING THAT THIS REGION SHOULD HARBOR SOME TUMOR SUPPRESSOR GENES (TSGS) CLOSELY RELATED TO NPC DEVELOPMENT. SEVERAL TSGS LOCATED AT 3P21.3, SUCH AS RASSF1A, LTF AND BLU, HAVE BEEN DEMONSTRATED TO BE INVOLVED IN NPC DEVELOPMENT. LARS2 (LEUCYL-TRNA SYNTHETASE 2, MITOCHONDRIAL) IS ANOTHER GENE LOCATED IN THE CHROMOSOME 3 COMMON ELIMINATED REGION-1 (C3CER1) AT 3P21.3. IN THIS STUDY, WE FOCUSSED ON THE EPIGENETIC AND GENETIC ALTERATIONS OF LARS2 IN NPC. THE MRNA EXPRESSION OF LARS2 WAS DETECTED IN 36 NPC AND 8 CHRONIC NASOPHARYNGITIS (NP) TISSUES BY SEMI-QUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (RT-PCR) AND REAL-TIME RT-PCR. SUBSEQUENTLY, THE MUTATION, ALLELIC LOSS, AND METHYLATION STATUS OF LARS2 WERE ANALYSED BY POLYMERASE CHAIN REACTION-SINGLE-STRAND CONFORMATION POLYMORPHISM (PCR-SSCP), HOMOZYGOUS DELETION (HD) ANALYSIS AND METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION IN PRIMARY NPC TISSUES. NO EXPRESSION OR DOWNREGULATION OF LARS2 WAS OBSERVED IN 78% OF PRIMARY NPC TISSUES. NO MUTATIONS, ASSESSED BY PCR-SSCP AND DNA SEQUENCING, WERE FOUND IN THE PROMOTER REGION AND EXON 1 OF LARS2 IN NPC TISSUES, WHEREAS HD WAS DETECTED IN 28% OF NPC SPECIMENS AT THE LARS2 LOCUS. IN ADDITION, HYPERMETHYLATION OF LARS2 WAS FOUND IN 64% OF NPC SAMPLES BUT ONLY IN 12.5% OF NP BIOPSIES. OUR DATA INDICATE THAT INACTIVATION OF LARS2 BY BOTH GENETIC AND EPIGENETIC MECHANISMS MAY BE A COMMON AND IMPORTANT EVENT IN THE CARCINOGENESIS OF NPC.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
5  3976  24 LONG-TERM DEFECTS OF NASAL EPITHELIUM BARRIER FUNCTIONS IN PATIENTS WITH NASOPHARYNGEAL CARCINOMA POST CHEMO-RADIOTHERAPY. BACKGROUND AND PURPOSE: CHRONIC AND RECURRENT UPPER RESPIRATORY TRACT INFECTION AND INFLAMMATION IS COMMON IN PATIENTS WITH NASOPHARYNGEAL CARCINOMA (NPC) POST CHEMO-RADIOTHERAPY (CRT). WHETHER IT IS DUE TO INTRINSIC (E.G., HOST-DEFENSE MECHANISMS OF THE EPITHELIUM), EPIGENETIC OR EXTRINSIC FACTORS IS NOT FULLY UNDERSTOOD. MATERIALS AND METHODS: TISSUE BIOPSIES OF THE MIDDLE TURBINATE (MT) AND INFERIOR TURBINATE (IT) FROM NPC PATIENTS AFTER CRT (MEAN OF 3 YEARS, N = 39) WERE COMPARED WITH THE IT BIOPSIES FROM HEALTHY SUBJECTS (N = 44). THE EPITHELIAL ULTRASTRUCTURE WAS EXAMINED BY TRANSMISSION ELECTRON MICROSCOPE (TEM). MRNA AND PROTEIN EXPRESSIONS OF EPITHELIAL STEM/PROGENITOR CELLS MARKERS, AS WELL MARKERS OF CELL PROLIFERATION AND DIFFERENTIATION MARKERS WAS ANALYZED. RESULTS: ABNORMAL EPITHELIAL ARCHITECTURE WAS OBSERVED IN ALL TISSUE SAMPLES OF NPC PATIENTS. SIGNIFICANTLY DECREASED EXPRESSION LEVELS OF MRNA AND PROTEIN LEVELS FOR P63 (BASAL CELLS), KI67 (CELL PROLIFERATION), P63(+)/KRT5(+) (EPITHELIAL STEM/PROGENITOR CELLS), MUC5AC AND MUC5B (SECRETARY PROTEINS FROM GOBLET CELLS), ALPHA-TUBULIN, BETA-TUBULIN AND TAP73 (CILIATED CELLS), DNAH5 AND DNAI1 AND RSPH4A (MICROTUBULE ASSEMBLIES OF MOTILE CILIA), FOXJ1 AND CP110 (CILIOGENESIS-ASSOCIATED MARKERS) WERE EVIDENT IN MT AND IT BIOPSIES FROM NPC PATIENTS WHEN COMPARED TO HEALTHY CONTROLS. CONCLUSION: CRT CAUSES LONG-TERM DEFECTS OF EPITHELIAL BARRIER FUNCTIONS AND INCREASES THE SUSCEPTIBILITY OF THESE PATIENTS TO UPPER RESPIRATORY TRACT INFECTION AND INFLAMMATION.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
6  5582  28 ROLE OF NITRATIVE AND OXIDATIVE DNA DAMAGE IN INFLAMMATION-RELATED CARCINOGENESIS. CHRONIC INFLAMMATION INDUCED BY BIOLOGICAL, CHEMICAL, AND PHYSICAL FACTORS HAS BEEN FOUND TO BE ASSOCIATED WITH THE INCREASED RISK OF CANCER IN VARIOUS ORGANS. WE REVEALED THAT INFECTIOUS AGENTS INCLUDING LIVER FLUKE, HELICOBACTER PYLORI, AND HUMAN PAPILLOMA VIRUS AND NONINFECTIOUS AGENTS SUCH AS ASBESTOS FIBER INDUCED INOS-DEPENDENT FORMATION OF 8-NITROGUANINE AND 8-OXO-7, 8-DIHYDRO-2'-DEOXYGUANOSINE (8-OXODG) IN CANCER TISSUES AND PRECANCEROUS REGIONS. OUR RESULTS WITH THE COLOCALIZATION OF PHOSPHORYLATED ATM AND GAMMA-H2AX WITH 8-OXODG AND 8-NITROGUANINE IN INFLAMMATION-RELATED CANCER TISSUES SUGGEST THAT DNA BASE DAMAGE LEADS TO DOUBLE-STRANDED BREAKS. IT IS INTERESTING FROM THE ASPECT OF GENETIC INSTABILITY. WE ALSO DEMONSTRATED IL-6-MODULATED INOS EXPRESSION VIA STAT3 AND EGFR IN EPSTEIN-BARR-VIRUS-ASSOCIATED NASOPHARYNGEAL CARCINOMA AND FOUND PROMOTER HYPERMETHYLATION IN SEVERAL TUMOR SUPPRESSOR GENES. SUCH EPIGENETIC ALTERATION MAY OCCUR BY CONTROLLING THE DNA METHYLATION THROUGH IL-6-MEDIATED JAK/STAT3 PATHWAYS. COLLECTIVELY, 8-NITROGUANINE WOULD BE A USEFUL BIOMARKER FOR PREDICTING THE RISK OF INFLAMMATION-RELATED CANCERS.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
7  3671  36 INFLAMMATION AND CANCER. INFECTION AND INFLAMMATION ACCOUNT FOR APPROXIMATELY 25% OF CANCER-CAUSING FACTORS. INFLAMMATION-RELATED CANCERS ARE CHARACTERIZED BY MUTAGENIC DNA LESIONS, SUCH AS 8-OXO-7,8-DIHYDRO-2'-DEOXYGUANOSINE (8-OXODG) AND 8-NITROGUANINE. OUR PREVIOUS STUDIES DEMONSTRATED THE FORMATION OF 8-OXODG AND 8-NITROGUANINE IN THE TISSUES OF CANCER AND PRECANCEROUS LESIONS DUE TO INFECTION (E.G., OPISTHORCHIS VIVERRINI-RELATED CHOLANGIOCARCINOMA, SCHISTOSOMA HAEMATOBIUM-ASSOCIATED BLADDER CANCER, HELICOBACTER PYLORI-INFECTED GASTRIC CANCER, HUMAN PAPILLOMAVIRUS-RELATED CERVICAL CANCER, EPSTEIN-BARR VIRUS-INFECTED NASOPHARYNGEAL CARCINOMA) AND PRO-INFLAMMATORY FACTORS (E.G., ASBESTOS, NANOMATERIALS, AND INFLAMMATORY DISEASES SUCH AS BARRETT'S ESOPHAGUS AND ORAL LEUKOPLAKIA). INTERESTINGLY, SEVERAL OF OUR STUDIES SUGGESTED THAT INFLAMMATION-ASSOCIATED DNA DAMAGE IN CANCER STEM-LIKE CELLS LEADS TO CANCER DEVELOPMENT WITH AGGRESSIVE CLINICAL FEATURES. REACTIVE OXYGEN/NITROGEN SPECIES FROM INFLAMMATION DAMAGE NOT ONLY DNA BUT ALSO OTHER BIOMACROMOLECULES, SUCH AS PROTEINS AND LIPIDS, RESULTING IN THEIR DYSFUNCTION. WE IDENTIFIED OXIDATIVELY DAMAGED PROTEINS IN CANCER TISSUES BY 2D OXYBLOT FOLLOWED BY MALDI-TOF/TOF. AS AN EXAMPLE, OXIDATIVELY DAMAGED TRANSFERRIN RELEASED IRON ION, WHICH MAY MEDIATE FENTON REACTIONS AND GENERATE ADDITIONAL REACTIVE OXYGEN SPECIES. DYSFUNCTION OF ANTI-OXIDATIVE PROTEINS DUE TO THIS DAMAGE MIGHT INCREASE OXIDATIVE STRESS. SUCH DAMAGE IN BIOMACROMOLECULES MAY FORM A VICIOUS CYCLE OF OXIDATIVE STRESS, LEADING TO CANCER DEVELOPMENT. EPIGENETIC ALTERATIONS SUCH AS DNA METHYLATION AND MICRORNA DYSREGULATION PLAY VITAL ROLES IN CARCINOGENESIS, ESPECIALLY IN INFLAMMATION-RELATED CANCERS. WE EXAMINED EPIGENETIC ALTERATIONS, DNA METHYLATION AND MICRORNA DYSREGULATION, IN EPSTEIN-BARR VIRUS-RELATED NASOPHARYNGEAL CARCINOMA IN THE ENDEMIC AREA OF SOUTHERN CHINA AND FOUND SEVERAL DIFFERENTIALLY METHYLATED TUMOR SUPPRESSOR GENE CANDIDATES BY USING A NEXT-GENERATION SEQUENCER. AMONG THESE CANDIDATES, WE REVEALED HIGHER METHYLATION RATES OF RAS-LIKE ESTROGEN-REGULATED GROWTH INHIBITOR (RERG) IN BIOPSY SPECIMENS OF NASOPHARYNGEAL CARCINOMA MORE CONVENIENTLY BY USING RESTRICTION ENZYME-BASED REAL-TIME PCR. THIS RESULT MAY HELP TO IMPROVE CANCER SCREENING STRATEGIES. WE PROFILED MICRORNAS OF NASOPHARYNGEAL CARCINOMA TISSUES USING MICROARRAYS. QUANTITATIVE RT-PCR ANALYSIS CONFIRMED THE CONCORDANT DOWNREGULATION OF MIR-497 IN CANCER TISSUES AND PLASMA, SUGGESTING THAT PLASMA MIR-497 COULD BE USED AS A DIAGNOSTIC BIOMARKER FOR NASOPHARYNGEAL CARCINOMA. CHRONIC INFLAMMATION PROMOTES GENETIC AND EPIGENETIC ABERRATIONS, WITH VARIOUS PATHOGENESES. THESE CHANGES MAY BE USEFUL BIOMARKERS IN LIQUID BIOPSY FOR EARLY DETECTION AND PREVENTION OF CANCER.	2018	

8  5274  14 PROMOTER METHYLATION OF P16 AND EDNRB GENE IN LEUKEMIA PATIENTS IN TAIWAN. BOTH EPIGENETIC AND GENETIC ALTERNATIONS ARE INVOLVED IN CANCER FORMATION. IN THIS STUDY, WE HAVE IDENTIFIED THE METHYLATION FREQUENCY OF P16 AND ENDOTHELIN RECEPTOR TYPE B (EDNRB) OF 26 LEUKEMIA PATIENTS AND 8 RANDOMLY SELECTED NORMAL BLOOD DONORS IN TAIWAN. PROMOTER METHYLATION OF P16 WAS DETECTED IN 85% OF ACUTE LYMPHOCYTIC LEUKEMIA (ALL), 83% IN ACUTE MYELOID LEUKEMIA (AML) WHEREAS NO METHYLATION WAS DETECTED IN CHRONIC MYELOID LEUKEMIA (CML) IN BLAST CRISIS. HYPERMETHYLATION OF EDNRB WAS OBSERVED IN 92% OF ALL, 75% AML AND 100% IN CML IN BLAST CRISIS. NO ABERRANT METHYLATION OF P16 AND EDNRB WAS FOUND IN 8 NORMAL BLOOD DONORS. TAKEN TOGETHER, ABERRANT METHYLATION OF P16 AND EDNRB WAS HIGHLY PREVALENT IN LEUKEMIA PATIENTS IN TAIWAN.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
9   878  33 CHRONIC CADMIUM EXPOSURE AGGRAVATES MALIGNANT PHENOTYPES OF NASOPHARYNGEAL CARCINOMA BY ACTIVATING THE WNT/BETA-CATENIN SIGNALING PATHWAY VIA HYPERMETHYLATION OF THE CASEIN KINASE 1ALPHA PROMOTER. BACKGROUND: OUR PREVIOUS STUDY HAS SHOWN THAT CADMIUM (CD) EXPOSURE IS NOT ONLY A RISK FACTOR FOR NASOPHARYNGEAL CARCINOMA (NPC), BUT ALSO CORRELATED WITH THE CLINICAL STAGE AND LYMPH NODE METASTASIS. HOWEVER, THE UNDERLYING MOLECULAR EVENTS OF CD INVOLVED IN NPC PROGRESSION REMAIN TO BE ELUCIDATED. PURPOSE: THE OBJECTIVE OF THIS STUDY WAS TO DECIPHER HOW CD IMPACTS THE MALIGNANT PHENOTYPES OF NPC CELLS. METHODS: NPC CELL LINES CNE-1 AND CNE-2 WERE CONTINUOUSLY EXPOSED WITH 1 MUM CD CHLORIDE FOR 10 WEEKS, DESIGNATING AS CHRONIC CD TREATED NPC CELLS (CCT-NPC). MTT ASSAY, COLONY FORMATION ASSAY AND XENOGRAFT TUMOR GROWTH WERE USED TO ASSESS CELL VIABILITY IN VITRO AND IN VIVO. TRANSWELL ASSAYS WERE PERFORMED TO DETECT CELL INVASION AND MIGRATION. THE PROTEIN LEVELS OF E-CADHERIN, N-CADHERIN, VIMENTIN AS WELL AS BETA-CATENIN AND CASEIN KINASE 1ALPHA(CK1ALPHA) WERE MEASURED BY WESTERN BLOT. IMMUNOFLUORESCENCE STAINING WAS USED TO OBSERVE THE DISTRIBUTION OF FILAMENT ACTIN (F-ACTIN), BETA-CATENIN AND CK1ALPHA. THE MRNA LEVELS OF DOWNSTREAM TARGET GENES OF BETA-CATENIN WERE DETECTED BY RT-PCR. WNT/BETA-CATENIN SIGNALING ACTIVITY WAS ASSESSED BY TOPFLASH/FOPFLASH DUAL LUCIFERASE REPORT SYSTEM. MS-PCR WAS USED TO DETECT THE METHYLATION STATUS OF CK1ALPHA. FINALLY, THE ACTIVATION OF WNT/BETA-CATENIN PATHWAY AND CELL BIOLOGICAL PROPERTIES WERE EXAMINED FOLLOWING TREATMENT OF CCT-NPC CELLS WITH 5-AZA-2-DEOXY-CYTIDINE(5-AZA-CDR). RESULTS: CCT-NPC CELLS SHOWED AN INCREASE IN CELL PROLIFERATION, COLONY FORMATION, INVASION AND MIGRATION COMPARED TO THE PARENTAL CELLS. CD ALSO INDUCED CYTOSKELETON REORGANIZATION AND EPITHELIAL-TO-MESENCHYMAL TRANSITION. UPREGULATION AND NUCLEAR TRANSLOCATION OF BETA-CATENIN AND INCREASED LUCIFERASE ACTIVITY ACCOMPANIED WITH TRANSCRIPTION OF DOWNSTREAM TARGET GENES WERE FOUND IN CCT-NPC CELLS. TREATMENT OF CCT-CNE1 CELLS WITH 5-AZA-CDR COULD REVERSE THE HYPERMETHYLATION OF CK1ALPHA AND ATTENUATE THE CELL MALIGNANCY. CONCLUSION: THESE RESULTS SUPPORT A ROLE FOR CHRONIC CD EXPOSURE AS A DRIVING FORCE FOR THE MALIGNANT PROGRESSION OF NPC VIA EPIGENETIC ACTIVATION OF THE WNT/BETA-CATENIN PATHWAY.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
10  4904  41 P16INK4A GENE ALTERATIONS ARE NOT A PROGNOSTIC INDICATOR FOR SURVIVAL IN PATIENTS WITH HEPATOCELLULAR CARCINOMA UNDERGOING CURATIVE HEPATECTOMY. BACKGROUND AND AIM: HEPATOCELLULAR CARCINOMA (HCC) IS A COMMON MALIGNANCY WORLDWIDE THAT IS HIGHLY ASSOCIATED WITH CHRONIC HEPATITIS B OR C INFECTION AND CIRRHOSIS. THE TUMOR SUPPRESSOR GENE P16INK4A IS AN IMPORTANT COMPONENT OF THE CELL CYCLE AND INACTIVATION OF THE GENE HAS BEEN FOUND IN A VARIETY OF HUMAN CANCERS. THE PRESENT STUDY WAS PERFORMED TO DETERMINE GENETIC AND EPIGENETIC ALTERATIONS IN THE P16INK4A TUMOR SUPPRESSOR GENE AND THE EFFECT OF THESE ON HCC PROGRESSION. METHODS: THE STATUS OF P16INK4A WAS EVALUATED IN 117 HCC TUMORAL NODULES AND 110 CORRESPONDING PERITUMORAL TISSUES BY LOSS OF HETEROZIGOSITY (LOH) AT THE 9P21-22 REGION, HOMOZYGOUS DELETIONS, SINGLE-STRAND CONFORMATION POLYMORPHISM-POLYMERASE CHAIN REACTION (PCR) MUTATIONAL ANALYSIS AND METHYLATION SPECIFIC PCR. RESULTS: THE MOST FREQUENT INACTIVATION MECHANISM WAS HYPERMETHYLATION OF THE PROMOTER REGION, WHICH WAS FOUND IN 63.2% OF THE TUMOR SAMPLES AND IN 28.2% OF THE PERITUMORAL SAMPLES. LOSS OF HETEROZYGOSITY AT THE 9P21 REGION WAS DETECTED IN 27.3% AND 10% OF TUMOR AND PERITUMORAL TISSUES, RESPECTIVELY. HOMOZYGOUS DELETIONS AND MUTATIONS WERE LESS COMMON EVENTS IN HEPATOCARCINOGENESIS. THE AUTHORS FOUND 5.9% OF THE TUMOR CASES WITH EXON 2 HOMOZYGOUS DELETIONS AND 8.6% WITH MUTATIONS. TWO POLYMORPHISMS WERE DETECTED, ONE AT CODON 148 (GCG --> ACG, ALA --> THR) IN THREE CASES AND THE OTHER IN EXON 3 AT 540 BP (34.2% OF THE SAMPLES). NO ASSOCIATION WAS FOUND BETWEEN INACTIVATION OF P16INK4A AND CLINICOPATHOLOGICAL CHARACTERISTICS OR PROGNOSIS. CONCLUSION: P16INK4A IS ALTERED FREQUENTLY AND EARLY IN HCC, BEING THE PREDOMINANT MECHANISM OF INACTIVATION PROMOTER HYPERMETHYLATION. THE PRESENT RESULTS SUGGEST THAT THE P16INK4A GENE PLAYS AN IMPORTANT ROLE IN THE PATHOGENESIS OF HCC.	2004	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
11  3231  29 HELICOBACTER PYLORI-INDUCED MODULATION OF THE PROMOTER METHYLATION OF WNT ANTAGONIST GENES IN GASTRIC CARCINOGENESIS. BACKGROUND: THIS STUDY AIMED TO INVESTIGATE THE CHANGES IN THE PROMOTER METHYLATION AND GENE EXPRESSION OF MULTIPLE WNT ANTAGONISTS BETWEEN THE CHRONIC INFECTION AND ERADICATION OF HELICOBACTER PYLORI (H. PYLORI) IN GASTRIC CARCINOGENESIS. METHODS: THE LEVELS OF METHYLATION AND CORRESPONDING MRNA EXPRESSION OF SEVEN WNT ANTAGONIST GENES (SFRP1, -2, -5, DKK1, -2, -3, WIF1) WERE COMPARED AMONG THE PATIENTS WITH H. PYLORI-POSITIVE GASTRIC CANCERS (GCS), AND H. PYLORI-POSITIVE AND H. PYLORI-NEGATIVE CONTROLS, BY QUANTITATIVE METHYLIGHT ASSAY AND REAL-TIME REVERSE TRANSCRIPTION (RT)-POLYMERASE CHAIN REACTION (PCR), RESPECTIVELY. THE CHANGES OF THE METHYLATION AND EXPRESSION LEVELS OF THE GENES WERE ALSO COMPARED BETWEEN THE H. PYLORI ERADICATION AND H. PYLORI-PERSISTENT GROUPS 1 YEAR AFTER ENDOSCOPIC RESECTION OF GCS. RESULTS: THE METHYLATION LEVELS OF SFRP AND DKK FAMILY GENES WERE SIGNIFICANTLY INCREASED IN THE PATIENTS WITH H. PYLORI-POSITIVE GCS AND FOLLOWED BY H. PYLORI-POSITIVE CONTROLS COMPARED WITH H. PYLORI-NEGATIVE CONTROLS (P < 0.001). SFRP1, -2, AND DKK3 GENE EXPRESSION WAS STEPWISE DOWNREGULATED FROM H. PYLORI-NEGATIVE CONTROLS, H. PYLORI-POSITIVE CONTROLS, AND TO H. PYLORI-POSITIVE GCS (P < 0.05). AMONG THE WNT ANTAGONISTS, ONLY THE DEGREES OF METHYLATION AND DOWNREGULATION OF DKK3 WERE SIGNIFICANTLY REDUCED AFTER H. PYLORI ERADICATION (P < 0.05). CONCLUSION: EPIGENETIC SILENCING OF SFRP AND DKK FAMILY GENES MAY FACILITATE THE FORMATION OF AN EPIGENETIC FIELD DURING H. PYLORI-ASSOCIATED GASTRIC CARCINOGENESIS. THE EPIGENETIC FIELD MAY NOT BE REVERSED EVEN AFTER H. PYLORI ERADICATION EXCEPT BY DKK3 METHYLATION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
12   672  26 BRAF, KRAS AND HELICOBACTER PYLORI EPIGENETIC CHANGES-ASSOCIATED CHRONIC GASTRITIS IN EGYPTIAN PATIENTS WITH AND WITHOUT GASTRIC CANCER. WE AIMED TO STUDY MLH1 AND MGMT METHYLATION STATUS IN HELICOBACTER PYLORI-ASSOCIATED CHRONIC GASTRITIS IN EGYPTIAN PATIENTS WITH AND WITHOUT GASTRIC CANCER. 39 PATIENTS WERE INCLUDED IN OUR STUDY. THEY WERE DIVIDED INTO 2 GROUPS; PATIENTS WITHOUT (GROUP I) AND WITH GASTRIC ADENOCARCINOMA (GROUP II). PATIENTS WERE SUBJECTED TO CLINICAL EXAMINATION, ABDOMINAL ULTRASOUND AND UPPER ENDOSCOPY FOR GASTRIC BIOPSY. BIOPSIES WERE SUBJECTED TO UREASE TEST, HISTOLOGICAL EXAMINATION, AND DNA PURIFICATION. H. PYLORI, BRAF, KRAS, MLH1 AND MGMT METHYLATION WERE ASSESSED BY QUANTITATIVE PCR. DNA SEQUENCING WAS PERFORMED TO ASSESS BRAF AND KRAS GENES MUTATION. QPCR OF H. PYLORI WAS SIGNIFICANTLY HIGHER IN PATIENTS WITH ADENOCARCINOMA (GROUP II) THAN THOSE WITHOUT ADENOCARCINOMA (GROUP I); WITH A P < 0.001 AS WELL AS IN PATIENTS WITH AGE ABOVE 50 YEARS WITH A P VALUE = 0.008. BY APPLYING LOGISTIC REGRESSION ANALYSIS IT WAS REPORTED THAT THE H. PYLORI QPCR IS A SIGNIFICANT PREDICTOR TO THE ADENOCARCINOMA WITH OR = 1.025 (95 % CI: 1. 002-1.048), WITH SENSITIVITY OF 90 % AND SPECIFICITY OF 100 %. ADENOCARCINOMA PATIENTS HAD A SIGNIFICANTLY HIGHER MEAN AGE AND LEVELS OF H. PYLORI, BRAF, K-RAS, METHYLATED MGMT AND METHYLATED MLH1 THAN THOSE OF GASTRITIS PATIENTS. DNA SEQUENCE ANALYSIS OF BRAF (CODON 12) AND KRAS (CODON 600) HAD GENES MUTATION IN GASTRIC ADENOCARCINOMA VERSUS CHRONIC GASTRITIS. CONCLUSION: H. PYLORI MAY CAUSE EPIGENETIC CHANGES PREDISPOSING THE PATIENTS TO CANCER STOMACH. ESTIMATION OF H. PYLORI BY QPCR CAN BE A GOOD PREDICTOR TO ADENOCARCINOMA. BRAF AND KRAS GENES MUTATION WERE REVELED IN GASTRITIS AND ADENOCARCINOMA PATIENTS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
13  6770  35 [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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
14  4231  32 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
15  6415  36 THE STUDY OF P16 AND P15 GENE METHYLATION IN HEAD AND NECK SQUAMOUS CELL CARCINOMA AND THEIR QUANTITATIVE EVALUATION IN PLASMA BY REAL-TIME PCR. EPIGENETIC SILENCING OF THE P16 AND P15 GENES BY PROMOTER METHYLATION ARE COMMONLY OBSERVED IN HUMAN EPITHELIAL MALIGNANCIES, INCLUDING HEAD AND NECK SQUAMOUS CELL CARCINOMAS (HNSCC). IN THIS STUDY, A METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) WAS USED TO EVALUATE THE METHYLATION STATUS OF THE P16 AND P15 GENES IN 73 HNSCC SURGICAL SPECIMENS. P16 AND P15 GENE METHYLATION WAS ALSO EXAMINED IN 29 PAIRED METASTATIC LYMPH NODES AND 29 PAIRED HISTOLOGICALLY, NORMAL RESECTION MARGIN MUCOSAE. THE QUANTITY OF CELL-FREE METHYLATED P16 AND P15 DNA IN THE PLASMA SAMPLES OF 20 HNSCC PATIENTS AND 24 HEALTHY CONTROLS WAS ALSO EXAMINED USING A FLUORESCENCE-BASED REAL-TIME PCR ASSAY. THE FREQUENCIES OF P16 AND P15 METHYLATION IN THE PRIMARY TUMOUR WERE 49% AND 60%, RESPECTIVELY. CONCORDANT METHYLATION OF P16 AND P15 IN TUMOUR SAMPLES AND METASTATIC LYMPH NODES WAS FOUND IN 59 AND 38% OF CASES, RESPECTIVELY. A SIGNIFICANTLY HIGHER PREVALENCE OF P15 METHYLATION WAS FOUND IN HISTOLOGICALLY-NORMAL SURGICAL MARGIN EPITHELIA OF HNSCC PATIENTS WITH CHRONIC SMOKING AND DRINKING HABITS COMPARED WITH NON-SMOKERS AND NON-DRINKERS. IN ADDITION, METHYLATED P16 AND P15 DNA LEVELS WERE SIGNIFICANTLY HIGHER IN THE PLASMA OF HNSCC PATIENTS (MEAN 56 COPIES/ML PLASMA AND 65 COPIES/ML PLASMA, RESPECTIVELY) COMPARED WITH NORMAL CONTROLS (MEAN 6 COPIES/ML PLASMA AND 16 COPIES/ML PLASMA, RESPECTIVELY). IN CONCLUSION, PROMOTER METHYLATION OF THE P16 AND P15 GENES IS INVOLVED IN THE PATHOGENESIS OF HNSCC AND MAY BE RELATED TO CHRONIC SMOKING AND DRINKING. THE DIFFERENTIAL LEVELS OF METHYLATED P16 AND P15 DNA IN PLASMA MIGHT BE POTENTIAL USEFUL MARKERS IN SCREENING HIGH-RISK POPULATIONS FOR EARLY HNSCC AND MONITORING THEIR TREATMENT RESPONSE.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
16  2842  40 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
17  2439  25 EPIGENETIC SILENCING OF THE MLH1 PROMOTER IN RELATION TO THE DEVELOPMENT OF GASTRIC CANCER AND ITS USE AS A BIOMARKER FOR PATIENTS WITH MICROSATELLITE INSTABILITY: A SYSTEMATIC ANALYSIS. BACKGROUND/AIMS: HUMAN MUTL HOMOLOG 1 (MLH1) PROMOTER METHYLATION WAS REPORTED IN GASTRIC CANCER (GC). THIS STUDY DETERMINED THE CLINICOPATHOLOGICAL, PROGNOSTIC, AND DIAGNOSTIC EFFECTS OF MLH1 PROMOTER METHYLATION IN GC. METHODS: THE COMBINED ODDS RATIO (OR) OR HAZARD RATIO (HR) AND THEIR CORRESPONDING 95% CONFIDENCE INTERVALS (95% CI) WERE CALCULATED. THE POOLED SENSITIVITY, SPECIFICITY, AND AREA UNDER THE CURVE (AUC) WERE ANALYZED. RESULTS: A TOTAL OF 4654 GC PATIENTS AND 3669 NON-MALIGNANT CONTROLS WERE IDENTIFIED IN THIS SYSTEMATIC ANALYSIS. MLH1 PROMOTER METHYLATION WAS SIGNIFICANTLY HIGHER IN GC SAMPLES THAN IN GASTRIC ADENOMAS, CHRONIC GASTRITIS, ADJACENT TISSUES, NORMAL GASTRIC MUCOSA, AND NORMAL HEALTHY BLOOD SAMPLES, BUT IT EXHIBITED A SIMILAR FREQUENCY IN GC VS. INTESTINAL METAPLASIA AND DYSPLASIA SAMPLES. MLH1 PROMOTER METHYLATION CORRELATED WITH AGE AND MICROSATELLITE INSTABILITY (MSI), BUT IT WAS NOT ASSOCIATED WITH GENDER, H. PYLORI INFECTION, SMOKING, DRINKING BEHAVIORS, PATHOLOGICAL HISTOLOGY, TUMOR DIFFERENTIATION, CLINICAL STAGE, LYMPH NODE STATUS, DISTANT METASTASIS, OR OVERALL SURVIVAL OF GC. MLH1 PROMOTER METHYLATION EXHIBITED A POOR SENSITIVITY VALUE (< 0.5) IN PATIENTS WITH GC COMPARED WITH ADJACENT TISSUES, GASTRIC ADENOMAS, CHRONIC GASTRITIS, NORMAL GASTRIC MUCOSA, AND NORMAL HEALTHY BLOOD SAMPLES. THE POOLED SENSITIVITY, SPECIFICITY, AND AUC OF MLH1 PROMOTER METHYLATION IN GC WITH MSI VS. GC WITH MICROSATELLITE STABILITY (MSS) SAMPLES WERE 0.64, 0.96, AND 0.90, RESPECTIVELY. CONCLUSIONS: OUR RESULTS SUGGEST THAT THE DETECTION OF MLH1 PROMOTER METHYLATION MAY BE A POTENTIAL PROGNOSTIC BIOMARKER FOR GC PATIENTS WITH MSI.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
18  4232  37 METHYLATION OF RUNX3 IN VARIOUS TYPES OF HUMAN CANCERS AND PREMALIGNANT STAGES OF GASTRIC CARCINOMA. ACCUMULATING EVIDENCE HAS IDENTIFIED A MECHANISM POTENTIALLY RESPONSIBLE FOR THE INACTIVATION OF TUMOR SUPPRESSOR GENES, NAMELY TRANSCRIPTIONAL SILENCING BY ABERRANT METHYLATION OF CPG ISLANDS. A PREVIOUS STUDY HAS SHOWN THE LOSS OF RUNX3 EXPRESSION, DUE TO ABERRANT METHYLATION OF ITS CPG ISLAND, IN GASTRIC CANCER CELL LINES, SUGGESTING THAT RUNX3 IS A TARGET FOR EPIGENETIC GENE SILENCING IN GASTRIC CARCINOGENESIS. HOWEVER, THERE ARE LIMITED DATA ON THE METHYLATION STATUS OF RUNX3 IN THE NEOPLASTIC AND NON-NEOPLASTIC TISSUES IN VARIOUS TYPES OF HUMAN CANCERS, INCLUDING GASTRIC CANCER. HERE, WE REPORT THAT 60% OF GASTRIC CANCER CELL LINES AND 64% OF PRIMARY GASTRIC CARCINOMAS (N=75) WERE METHYLATED AT THE RUNX3 CPG ISLAND. RUNX3 METHYLATION WAS ALSO DETECTED IN HEPATOCELLULAR CARCINOMAS (73%, N=48), LARYNX CANCERS (62%, N=37), LUNG CANCERS (46%, N=24), BREAST CANCERS (25%, N=25), PROSTATE CANCERS (23%, N=44), ENDOMETRIAL CANCERS (12.5%, N=24), COLON CANCERS (4.9%, N=61) AND UTERINE CERVICAL CANCERS (2.5%, N=40), SHOWING THAT RUNX3 METHYLATION IS NOT RESTRICTED TO GASTRIC CANCER. INTERESTINGLY, THE RUNX3 METHYLATION WAS ESPECIALLY FREQUENT IN TUMORS FROM TISSUES OF A FOREGUT DERIVATIVE, THAT IS, THE STOMACH, LIVER, LARYNX AND LUNG. NEXT, THE METHYLATION STATUS OF RUNX3 IN VARIOUS NON-NEOPLASTIC TISSUES WAS EXAMINED, INCLUDING THE PREMALIGNANT LESIONS OF GASTRIC CARCINOMAS. THE RUNX3 METHYLATION WAS FOUND IN 8.1% OF CHRONIC GASTRITIS (N=99), 28.1% OF INTESTINAL METAPLASIA (N=32), 27.3% OF GASTRIC ADENOMAS (N=77) AND 64% OF GASTRIC CARCINOMAS (N=75), BUT NOT IN CHRONIC HEPATITIS B, NORMAL PROSTATE AND COLON MUCOSA, EVEN THOUGH IN CASES OF CHRONIC HEPATITIS, THE METHYLATION FREQUENCY OF ITS NEOPLASTIC TISSUES WAS VERY HIGH. IN CONCLUSION, RUNX3 METHYLATION IS FREQUENTLY FOUND IN HUMAN CANCERS, INCLUDING GASTRIC CANCER, AND IS MOSTLY CANCER SPECIFIC, WITH THE EXCEPTION OF THE STOMACH, AND THUS, MIGHT BE USEFUL AS A POTENTIAL DIAGNOSTIC BIOMARKER OF CANCER.	2004	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
19  1342  37 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
20  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