OBJECTIVETo correlate the clinical characteristics with mutations of the STK11 and FHIT genes in 16 patients with Peutz-Jeghers syndrome (PJS).

METHODSPotential mutations in the coding regions and flanking sequences of the STK11 and FHIT genes were detected with PCR and Sanger sequencing.

RESULTSOf the 16 patients with PJS, 8 had novel mutations in the coding region of the STK11 gene, 1 had a previously reported mutation. 1 carried a mutation in the exon 10 of the FHIT gene, which is a non-coding region. None of the mutations was detected in the immediate family members. None of the patients with STK11 gene mutations had mutation in the FHIT gene. The mutation rate of the STK11 gene among patients with PJS was 56.25%.

CONCLUSIONMutations of the STK11 gene are the major cause of PJS. Few such patients had mutations of the FHIT gene. Mutations of the FHIT gene may play a part in the pathogenesis of PJS.

Acid Anhydride Hydrolases ; genetics ; Adolescent ; Adult ; Base Sequence ; Child ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; Neoplasm Proteins ; genetics ; Pedigree ; Peutz-Jeghers Syndrome ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; Young Adult

OBJECTIVETo investigate the promoter methylation of p16, FHIT and RASSF1A gene and telomere damage in the workers exposed to coal tar pitch, and to explore the effective biomarker of occupational exposure to coal tar pitch.

METHODS180 cases of workers exposed to coal tar pitch in a certain carbon plant named as exposure group, and 145 healthy cases with a medical examination in the first affiliated hospital of Zhengzhou University were selected as control group. Relative telomere length in peripheral blood DNA was detected using real-time quantitative PCR, and the promoter methylation rate of p16, RASSF1A and FHIT gene in peripheral blood DNA were determined by real-time quantitative methylation specific PCR. The relative telomere length and gene promoter methylation in two groups were compared, and influencing factors were analyzed.

RESULTSRelative telomere length in exposed group was lower than that in the control group, and the difference was statistically significant (Z = -5.395, P < 0.001). There was no significant difference in the promoter methylation rate of p16, FHIT and RASSF1A gene between the two groups (P > 0.05). Stratification analysis by gender, age, and smoking, we found that when the age was less than or equal to 40, the promoter methylation rate of p16 in exposed group was more than that in control group, and the difference was statistically significant (Z = -1.914, P = 0.011).

CONCLUSIONOccupational exposure to coal tar pitch may induce leukocyte DNA telomere length of human peripheral blood shortened, and may not change the promoter methylation rates of p16, FHIT and RASSF1A gene.

Acid Anhydride Hydrolases ; genetics ; Coal Tar ; adverse effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; Humans ; Leukocytes ; drug effects ; Neoplasm Proteins ; genetics ; Occupational Exposure ; adverse effects ; Promoter Regions, Genetic ; Telomere ; drug effects ; ultrastructure ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation, expression of mRNA and protein of fragile histidine triad (FHIT)gene in cervical cancer cells.

METHODSCervical cancer cell lines including CaSki (HPV16-positive) and C33A (HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR (MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR, respectively.

RESULTSFolate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r = 0.98, P < 0.001; CaSki:r = 0.98, P < 0.001) and the apoptosis rate increased (C33A:r = 0.98, P < 0.001; CaSki:r = 0.99, P < 0.001) along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1 µg/ml and 10 µg/ml, partially positive at 100 µg/ml and 250 µg/ml, but negative at 500 µg/ml and 1 000 µg/ml in both C33A and CaSki cells. Comparing with the control group, the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells, and showing that the FHIT gene mRNA expression (C33A:r = 0.96, P < 0.001; CaSki:r = 0.94, P < 0.001) and protein expression (C33A:r = 0.96, P < 0.001; CaSki:r = 0.97, P < 0.001) both increased along with the increase of folate concentration.

CONCLUSIONOur findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro, so would reverse the aberration mRNA and protein expression of FHIT gene.

Acid Anhydride Hydrolases ; genetics ; metabolism ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Culture Media ; chemistry ; DNA Methylation ; Female ; Folic Acid ; pharmacology ; Humans ; Neoplasm Proteins ; genetics ; metabolism ; Uterine Cervical Neoplasms ; genetics ; pathology

This study was aimed to detect the methylation status of FHIT gene promoter region in the DNA from plasma of patients with myelodysplastic syndrome (MDS), and to investigate the demethylating effect of decitabine. Methylation-specific PCR method was used to detect the methylation status of FHIT gene promoter region in the DNA from plasma of 4 patients with MDS before and after treatment with decitabine plus semis CAG therapy (among them, 1 case of newly diagnosed MDS, 3 cases progressed into acute leukemia). The results indicated that 3 cases were found to have an increased methylation in the promoter region. After treatment with decitabine plus semis CAG, increased methylation was reversed in 2 cases. In 4 cases, 2 cases displayed clinical response. It is concluded that FHIT gene hypermethylation is associated with MDS pathogenesis. Decitabine has demethylating effect on the FHIT gene hypermethylation of plasma from MDS patients. Detecting the methylation status of FHIT gene in DNA from plasma may play a role in MDS auxiliary diagnosis or prognosis.
Acid Anhydride Hydrolases ; genetics ; Adult ; Aged ; Azacitidine ; analogs & derivatives ; therapeutic use ; DNA ; blood ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; drug therapy ; Neoplasm Proteins ; genetics ; Promoter Regions, Genetic

Acid Anhydride Hydrolases ; genetics ; metabolism ; Animals ; Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Biomarkers, Tumor ; analysis ; Carcinoma, Squamous Cell ; diagnosis ; genetics ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA Methylation ; drug effects ; Genes, p16 ; Humans ; Laryngeal Neoplasms ; diagnosis ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; genetics ; metabolism ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo investigate the effect of fragile histidine triad (FHIT) gene transfection on human colorectal cancer cell line SW480 through up-regulation of caspase-8 expression.

METHODSThe eukaryotic expression plasmid containing FHIT, pRc/CMV2-FHIT was prepared and purified, and then identified by restrictive enzyme digestion. pRc/CMV2-FHIT was transfected into SW480 cells, and positive cell clones (SW480-FHIT, study group) were selected and amplified. Empty plasmid-transfected SW480 cells(SW480-pRc/CMV2, negative control) and normal SW480 cells (bland control) were used as control. Methyl thiazolyl tetrazolium (MTT) assay was used to test the changes in the proliferation of SW480 cells. Cell-cycle kinetics and apoptosis were analyzed by flow cytometry (FCM). The changes of pro-caspase-8, caspase-8 mRNA and caspase-8 relative activity were analyzed by Western blot, semi-quantitative RT-PCR and colorimetric assay with pan labeled substrate, respectively.

RESULTSAt 96 hours after transfection, cell inhibition rates of the study group and the negative control group were 71.7% and 16.9%. G0/G1 ratio was (63.2±3.5)% and (50.6±2.1)%, optical density of caspase-8 mRNA band 107 and 41, and relative activity of caspase-8 0.43 and 0.25, respectively. All the differences above were statistically significant (P<0.05). When FHIT inhibitor was added, the relative activity of caspase-8 decreased to 0.22, comparable to that in the control group.

CONCLUSIONSFHIT gene transfection can significantly inhibit the proliferation and induce G0/G1 arrest in human colon cancer cell line SW480. The mechanism is related to the up-regulation of caspase-8 expression.

Acid Anhydride Hydrolases ; genetics ; Apoptosis ; Caspase 8 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; genetics ; pathology ; Humans ; Neoplasm Proteins ; genetics ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo study the relationship between FHIT and WWOX expression and clinicopathological features in hepatocellular carcinoma (HCC).

METHODThe expression of FHIT and WWOX were determined by immunohistochemistry in 142 patients with HCC.

RESULTSAbsent or reduced FHIT and WWOX expression was observed in 68.3% and 77.5% of HCCs, respectively. The expression of FHIT was significantly correlated with that of WWOX (P < 0.01), and progressive loss of FHIT and WWOX expression were observed as tumor differentiation decreased and tumor grade increased (P < 0.05). Absent/reduced FHIT and WWOX expression was associated with tumor invasion and metastasis (P < 0.05). In addition, the expression of FHIT and WWOX in HCC with recrudesce were lower than that in those without recrudesce (P < 0.05).

CONCLUSIONSAbsent/reduced FHIT and WWOX expression is associated with poor prognosis.

Acid Anhydride Hydrolases ; genetics ; Adolescent ; Adult ; Aged ; Carcinoma, Hepatocellular ; genetics ; pathology ; Female ; Gene Expression Profiling ; Humans ; Liver Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; Oxidoreductases ; genetics ; Prognosis ; Tumor Suppressor Proteins ; genetics ; WW Domain-Containing Oxidoreductase ; Young Adult

OBJECTIVE: To determine the role of fragile histidine triad (FHIT) and MDM2 in carcinogenesis of oral submucous fibrosis (OSF). METHODS: The expression of FHIT and MDM2 was examined by immunohistochemical S-P method in 44 OSF cases, 15 canceration tissues of OSF, and 10 normal oral mucosa tissues. RESULTS: The expression of FHIT was positive in the normal oral mucosa epithelium. The positive expression of FHIT decreased in the OSF and canceration tissues of the OSF.The rate of FHIT positive expression was significantly lower in canceration tissues of OSF than that of the OSF (P < 0.05). The expression of MDM2 was negative in normal oral mucosa epithelium. The positive expression of MDM2 increased in the OSF and canceration tissues of the OSF, and the rate of MDM2 positive expression was significantly higher in the canceration tissues of OSF than that of the OSF (P < 0.05). CONCLUSION The loss of FHIT and over-expression of MDM2 may play an important role in the carcinogenesis of OSF.
Acid Anhydride Hydrolases ; genetics ; metabolism ; Female ; Humans ; Immunohistochemistry ; Male ; Mouth Mucosa ; metabolism ; Mouth Neoplasms ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Oral Submucous Fibrosis ; metabolism ; Proto-Oncogene Proteins c-mdm2 ; genetics ; metabolism

Cholangiocarcinoma (CC) is an intrahepatic bile duct carcinoma with a high mortality rate and a poor prognosis. Sarcomatous change/epithelial mesenchymal transition (EMT) of CC frequently leads to aggressive intrahepatic spread and metastasis. The aim of this study was to identify the genetic alterations and gene expression pattern that might be associated with the sarcomatous change in CC. Previously, we established 4 human CC cell lines (SCK, JCK1, Cho-CK, and Choi-CK). In the present study, we characterized a typical sarcomatoid phenotype of SCK, and classified the other cell lines according to tumor cell differentiation (a poorly differentiated JCK, a moderately differentiated Cho-CK, and a well differentiated Choi-CK cells), both morphologically and immunocytologically. We further analyzed the genetic alterations of two tumor suppressor genes (p53 and FHIT) and the expression of Fas/FasL gene, well known CC-related receptor and its ligand, in these four CC cell lines. The deletion mutation of p53 was found in the sarcomatoid SCK cells. These cells expressed much less Fas/FasL mRNAs than did the other ordinary CC cells. We further characterize the gene expression pattern that is involved in the sarcomatous progression of CC, using cDNA microarrays that contained 18,688 genes. Comparison of the expression patterns between the sarcomatoid SCK cells and the differentiated Choi-CK cells enabled us to identify 260 genes and 247 genes that were significantly over-expressed and under-expressed, respectively. Northern blotting of the 14 randomly selected genes verified the microarray data, including the differential expressions of the LGALS1, TGFBI, CES1, LDHB, UCHL1, ASPH, VDAC1, VIL2, CCND2, S100P, CALB1, MAL2, GPX1, and ANXA8 mRNAs. Immunohistochemistry also revealed in part the differential expressions of these gene proteins. These results suggest that those genetic and gene expression alterations may be relevant to the sarcomatous change/EMT in CC cells.
Acid Anhydride Hydrolases/genetics ; Animals ; Cell Line, Tumor ; Cholangiocarcinoma/*genetics ; Female ; *Gene Expression Profiling ; Humans ; Mice ; Mice, Inbred BALB C ; Mutation ; Neoplasm Proteins/genetics ; Oligonucleotide Array Sequence Analysis ; Sarcoma/*genetics ; Tumor Suppressor Protein p53/genetics

Acid Anhydride Hydrolases ; metabolism ; Apoptosis ; Biomarkers, Tumor ; metabolism ; Caspases ; metabolism ; Cell Adhesion Molecules ; metabolism ; Chromosome Deletion ; Drug Delivery Systems ; GPI-Linked Proteins ; metabolism ; Humans ; Hyaluronoglucosaminidase ; metabolism ; In Situ Hybridization, Fluorescence ; methods ; Lung Neoplasms ; diagnosis ; drug therapy ; genetics ; metabolism ; Neoplasm Proteins ; metabolism ; Neoplasm Staging ; Receptor, Epidermal Growth Factor ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism