Objective: To explore the relationship between abnormal expression of fragile histidine triad (FHIT) gene and methyl-CpG-binding protein 2 (MeCP2) as well as their interaction on cervical cancerization. Methods: A total of 73 patients with cervical squamous cell carcinoma (SCC), 113 patients with cervical intraepithelial neoplasia (CIN Ⅰ, n=45; CINⅡ/Ⅲ, n=68) and 60 women with normal cervix (NC) were included in the study. Real time PCR and Western blot were performed to detect the expression levels of mRNA and protein about FHIT and MeCP2, respectively. The methylation status of FHIT gene CpG island was tested by methylation-specifc PCR (MSP). Kruskal-Wallis H test, χ(2) test, trend χ(2) test and Spearman correlation analysis were conducted with software SPSS 20.0. The interaction was evaluated by generalized multifactor dimensionality reduction (GMDR) model. Results: With the deterioration of cervical lesion, the methylation rates of FHIT gene CpG island (χ(2)=18.64, P<0.001; trend χ(2)=18.08, P<0.001) increased gradually, while the expression levels of FHIT mRNA (H=27.32, P<0.001; trend χ(2)=12.65, P<0.001) and protein (H=47.10, P<0.001; trend χ(2)=29.79, P<0.001) decreased gradually. There was a negative correlation between the methylation rates of FHIT gene CpG island and the expression level of FHIT protein (r=-0.226, P<0.001). The levels of MeCP2 mRNA (H=26.19, P<0.001; trend χ(2)=11.81, P=0.001) and protein (H=69.02, P<0.001; trend χ(2)=47.44, P<0.001) increased gradually with the aggravation of cervical lesions. There was a positive correlation between the expression level of MeCP2 protein and the FHIT mRNA Ct ratio (r=0.254, P<0.001). Expression of proteins were negatively correlated between MeCP2 and FHIT (r=-0.213, P=0.001). The results analyzed by GMDR model showed that there were interactions among high MeCP2 protein expression, the CpG island methylation of FHIT and mRNA and protein expression in CINⅡ/Ⅲ group, and among high MeCP2 mRNA and protein expression, the CpG island methylation of FHIT and low mRNA and protein expression in SCC group. Conclusion: High expression of MeCP2 mRNA and protein, the CpG island methylation and low mRNA and protein expression of FHIT could increase the risk of cervical carcinogenesis, and there might be a synergistic effect on cervical carcinogenesis.
Acid Anhydride Hydrolases/metabolism* ; Carcinoma, Squamous Cell/pathology* ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Methyl-CpG-Binding Protein 2/metabolism* ; Neoplasm Proteins/metabolism* ; Polymerase Chain Reaction/methods* ; RNA, Messenger ; Uterine Cervical Neoplasms/pathology* ; Uterine Cervical Dysplasia/pathology*

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

BACKGROUNDIt is widely recognized that the diagnosis of parathyroid carcinoma (PC) is often difficult because of the overlap of characteristics between malignant and benign parathyroid tumors, especially at an early stage. Our study aimed to investigate the differential expression of Ki-67, galectin-3, fragile histidine triad (FHIT) gene, and parafibromin in PC, parathyroid adenoma (PA), parathyroid hyperplasia (PH), and normal parathyroid (NP) tissues; then to assess these expression values for use in differential diagnosis of malignant and benign parathyroid tumors.

METHODSData of 15 cases with PC, 19 PAs, and 8 PHs were retrospectively analyzed for their clinical characteristics. The expression of Ki-67, galectin-3, FHIT, and parafibromin were detected via immunohistochemistry in the above-mentioned specimens and 6 NPs as control.

RESULTSComplete loss of parafibromin expression was seen in 9 of 15 (60%) carcinomas, and all normal parathyroid tissues and parathyroid benign tumors stained positive for parafibromin except for one (4%) adenoma. Galectin-3 staining was positive in 11 of 15 (73%) carcinomas, 5 of 19 (26%) adenomas, 1 of 8 (12%) hyperplasias, and 0 of 6 normal tissues. The Ki-67 proliferative index was high in 4 of 15 (27%) carcinomas, 1 of 19 (5%) adenomas, and none of the hyperplasia or normal tissues. FHIT expression did not differ appreciably among the tumor types. The combination of overexpression of galectin-3 or loss of parafibromin increased sensitivity for PC to 87%, while the specificity of both positive galectin-3 and positive Ki-67 could reach 100%.

CONCLUSIONSThese data suggested that loss of parafibromin and overexpression of galectin-3 and Ki-67 might help to distinguish parathyroid carcinoma from other parathyroid tumors. And the combination of two or three of these markers might produce better sensitivity and/or specificity for the diagnosis of parathyroid carcinoma.

Acid Anhydride Hydrolases ; metabolism ; Galectin 3 ; metabolism ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Neoplasm Proteins ; metabolism ; Parathyroid Neoplasms ; metabolism ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo investigate the expressions of the FHIT and PTEN genes and their significance in prostate cancer.

METHODSThe expressions of FHIT and PTEN were detected in 85 cases of prostate cancer and 30 cases of benign prostatic nodular hyperplasia by immunohistochemistry of PV-6000.

RESULTSThe positive expression rates of FHIT and PTEN were 34.1% and 42.4% in prostate cancer, significantly lower than 96.7% and 90.0% in benign prostatic nodular hyperplasia (P <0.01). Statistically significant differences were found in the positive expression rates of FHIT and PTEN among different Gleason grades, 44.4% and 55.6% in well differentiated, 38.9% and 44.4% in moderately differentiated, and 25.0% and 37.5% in lowly differentiated prostate cancer (P <0.05). But the expression of FHIT.

CONCLUSIONFHIT and PTEN may play a certain role in the was not correlated with that of PTEN in the prostate cancer tissue (P >0.05). development, progression and infiltration of prostate cancer.

Acid Anhydride Hydrolases ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Aged ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology

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 expression and significance of fragile histidine triad (FHIT) and Ki-67 in transformed epithelial cells induced by Yunnan tin mine dust.

METHODSEvery second generation of immortalized human bronchial epithelial cells (BEAS-2B) and human embryo lung fibroblasts (WI-38) were exposed to 100 µg/ml Yunnan tin mine dust for 72 h, until the ninth generation. The cells were subsequently co-cultured from the 11th generation. Experimental setup: B group, B (W) group, B (W 100) group, B100 group, B100 (W) group, B100 (W100) group. The expressions of FHIT and Ki-67 in epithelial cells were determined by the method of immunocytochemistry at the 16th, 26th and 36th generation. The percentage of Ki-67 positive cells was calculated as proliferation index.

RESULTSThe expression of FHIT was observed in BEAS-2B cells. The expression levels of FHIT among B group, B (W) group and B (W 100) group had not instinctive difference. At the 16th generation, the expression of FHIT in the B100 group was decreased compared with that in the B group and the expression of FHIT between B100 (W) group and B100 (W100) group was lower than that in the B100 group. At the 26th generation, the expression of FHIT was decreased compared with that at the 16th generation in the B100, B100 (W) and B100 (W100) groups. However, At the 36th generation, positive expression were observed again in the B100, B100 (W) and B100 (W100) groups and the expression levels were in incremental order. At the 16th, 26th and 36th generation, the proliferation indexes of B group, B (W) group and B (W 100) group were all < 3%. The proliferation indexes of B100, B100 (W) and B100 (W100) were increased step by step with the generation elongation.

CONCLUSIONSFHIT could be a target at which Yunnan tin mine dust induces transformation of BEAS-2B cells. The proliferation activation of BEAS-2B cells can be improved by Yunnan tin mine dust.

Acid Anhydride Hydrolases ; metabolism ; Cell Line ; Cell Transdifferentiation ; China ; Dust ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Ki-67 Antigen ; metabolism ; Lung ; cytology ; Neoplasm Proteins ; metabolism ; Tin ; toxicity

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

OBJECTIVETo detect the expression of fragile histidine triad in endometriosis and investigate the pathogenesis of endometriosis.

METHODSimmunohistochemistry was used to examine the expression of Fhit in the eutopic and ectopic endometria of 58 patients with endometriosis and in the endometria in 15 patients with hysteromyoma.

RESULTSThe intensity of Fhit expression decreased in the order of normal endometrium, eutopic endometrium in endometriosis group, and ectopic endometrium. In patients with endometriosis, Fhit expression in the eutopic and ectopic endometria in proliferative phase showed no significant difference from that in secretory phase (P>0.05). Fhit expression in the ectopic endometrium showed significant difference between different r-AFS stages. MOD of ectopic endometrium in stages I-II was significantly higher than that in stages III-IV (P<0.05), but Fhit expression in the eutopic endometrium showed no significant difference (P>0.05). MOD of ovarian endometriosis showed no difference with that of adenomyosis.

CONCLUSIONSFhit may play an important role in the development of endometriosis.

Acid Anhydride Hydrolases ; metabolism ; Adult ; Endometriosis ; metabolism ; pathology ; Endometrium ; metabolism ; Female ; Humans ; Middle Aged ; Neoplasm Proteins ; metabolism

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