Acid Anhydride Hydrolases ; Carcinoma, Hepatocellular ; etiology ; genetics ; Humans ; Liver Neoplasms ; etiology ; genetics ; Mutation ; Neoplasm Proteins ; genetics ; RNA, Messenger ; analysis

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

OBJECTIVETo investigate the suppression effect of exogenous fragile histidine triad (FHIT) gene on biological property of MEC-1 cells.

METHODSIn order to study the FHIT function in MEC-1 cells, wild-type FHIT gene was transferred into mucoepidermoid carcinoma MEC-1 cells. The proliferation and kinetics, cell cycle, clonal forming rate, and apoptosis of MEC-1 cells, before and after FHIT gene transfection in vitro, and tumor loci formed on mice after injection of transferred MEC-1 cells in vivo were observed by immunochemical staining, flow cytometry analysis, and so on.

RESULTSIt can be seen that exogenous FHIT gene transfer could significantly inhibit the proliferation and reduce the kinetics of MEC-1 cells in vitro, prolong DT from (21.03+/-0.41) h to (26.86+/-0.71) h, and also keep less cells in cell cycle phase S, whilst more cells in phase G1, Additionally, the exogenous FHIT was found to be able to remarkably suppress MEC-1 cells of forming tumor loci in nude mice by lessen tumor weight, and promote higher differentiation of MEC-1 cells to be mucous cells.

CONCLUSIONExogenous FHIT gene could suppress the proliferation, tumorigenicity and improve the differentiation of MEC-1 cells, in vitro and in vivo.

Acid Anhydride Hydrolases ; Animals ; Apoptosis ; Carcinoma, Mucoepidermoid ; Cell Cycle ; Cell Line, Tumor ; Histidine ; Humans ; Mice ; Mice, Nude ; Neoplasm Proteins ; Transfection

OBJECTIVETo investigate the expression of fragile histidine triad (FHIT) gene protein, FHIT and the possible relationship between FHIT expression and clinicopathological indices in colorectal carcinoma.

METHODSDetecting FHIT protein expression in 60 cases of formalin-fixed, paraffin-embedded colorectal carcinoma by citrate-microwave-SP immunohistochemical method, and analyzing its relationship to histological grade, Dukes' stage and 5-year survival rate.

RESULTS55% of the carcinomas showed a marked loss or absence of FHIT expression compared with their matched normal mucosa. Carcinomas with reduced expression of FHIT correlated with their histological grade, Dukes' stage (P < 0.05) and 5-year survival rate. The distribution of decreased expression of FHIT was 7/16 in grade I carcinoma, 14/30 in grade II, 12/14 in grade III, respectively. The correlation between decreased expression of FHIT and Dukes' staging was 5/11 in stage A, 12/28 in stage B, and 16/21 in stage C. The difference between stage A, B with no lymph nodes metastases and the stage C with lymph nodes metastases was of significance (P < 0.05). The follow-up data of 39 cases showed that in the 5-year survival group, 13/25 were of the low FHIT expression carcinomas, while in 5-year deceased group 12/14 were of the low FHIT expression carcinomas (P < 0.05).

CONCLUSIONSThe reduced expression of FHIT may be associated with decreasing differentiation, metastasis and 5-year survival rate in colorectal carcinoma. It is suggested that decreased FHIT expression plays an important role in the development and progression of the tumor, and thus may become a new prognostic marker in colorectal carcinoma.

Acid Anhydride Hydrolases ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Neoplasm Proteins ; biosynthesis ; Neoplasm Staging ; Survival Analysis

BACKGROUND/AIMS: The fragile histidine triad (FHIT) gene located at chromosome 3p14.2, is a candidate tumor suppressor gene often involved in various tumors. Homozygous deletions, lack or reduced expression of FHIT protein, and alteration of its transcription were frequently observed in several types of primary human cancers and cell lines. In the present study, we examined the expression profiles of aberrant FHIT transcripts to explore the role of FHIT gene in gastric carcinogenesis. METHODS: In 6 gastric cancer cell lines, nested reverse transcription-polymerase chain reaction (RT-PCR) and cDNA sequence analyses were performed to detect and characterize the aberrant FHIT transcripts. RESULTS: In addition to the wild-type FHIT transcript, small-sized transcripts with various numbers and lengths were observed in all of the cell lines examined. cDNA sequence analysis confirmed that different types of truncated transcripts included exonic deletions, insertions of intron 5 sequences between exons, and combinations of both. Most of these transcripts lacked exon 5 in which translation initiation codon is located. Aberrant transcripts with partial exonic deletions due to activation of cryptic splice sites were also observed in 5 cell lines. Additionally, multi-step splice patterns indicative of additional downstream processing, were observed in several cancer lines. CONCLUSIONS: These results suggest that the aberrant FHIT transcripts in gastric cancer cell lines results from faulty splicing, including exon skipping, selection of cryptic splice site, and additional downstream splice processing.
*Acid Anhydride Hydrolases ; Cell Line, Tumor ; Codon, Initiator/genetics ; DNA, Complementary/genetics ; Genes, Tumor Suppressor ; Humans ; Neoplasm Proteins/*genetics ; *Sequence Analysis, DNA ; Stomach Neoplasms/*genetics ; *Transcription, Genetic

OBJECTIVETo detect the alteration of fragile histidine triad (FHIT) gene and p16 gene during malignant transformation of immortal human bronchial epithelial cell line (16HBE) induced by crystalline nickel sulfide, and study the molecular mechanism of nickel carcinogenesis.

METHODSMalignant transformed cells and tumorigenic cells were examined for the alteration of FHIT gene and p16 gene by RT-PCR, DNA sequencing and silver staining PCR-SSCP.

RESULTSCompared with those of control 16HBE, neither mutation of exon2 or exon2-3, abnormal expression in p16 gene nor mutation of FHIT exon5, 6, 7 and 8, exon1-4 or exon5-9 were observed in transformed cells and tumorigenic cells. But aberrant transcript or FHIT gene expression loss were observed in transformed cells and tumorigenic cells. One of the aberrant transcripts in FHIT gene, the deletion of exon6, exon7 and exon8 and an insertion of 36 bp sequence replacing exon6-8, was confirmed by sequencing.

CONCLUSIONFHIT gene, not p16 gene, could play a definite role in nickel carcinogenesis. Alterations of FHIT gene induced by crystalline NiS could be a molecular event associated with carcinogen, chromosome fragile site instability and cell malignant transformation, and FHIT gene could be one of the important target genes activated by exotic carcinogens.

Acid Anhydride Hydrolases ; Base Sequence ; Cell Transformation, Neoplastic ; chemically induced ; metabolism ; Cells, Cultured ; Gene Expression ; Genes, p16 ; physiology ; Humans ; Molecular Sequence Data ; Neoplasm Proteins ; genetics ; metabolism ; Nickel ; pharmacology

FHIT (fragile histidine triad) gene at chromosome 3p14.2 usually expresses at a very low level in human tissue and cells. A high frequency of abnormalities in FHIT gene has been demonstrated in various cancers. FHIT is proposed as a putative tumor-suppressor gene. To evaluate the expression of the FHIT gene in various leukemias, bone marrow or peripheral blood samples from 98 leukemia patients were tested by RT-PCR: 38 from patients with AML-[M(2)(9), M(3)(12), M(4)(8), M(5)(9)], 16 with ALL, and 34 with CML-[CP(20), AP(4), BC(10)] of various FAB types, as well as 10 patients with other hematological malignancies. To detect a deletion in sequencing the FHIT gene, the representative aberrant PCR products were cloned and then sequenced. The results showed that 22/38 (58%) patients with AML, 9/16 (56%) patients with ALL and 19/34 (56%) patients with CML were detectable of aberrant FHIT mRNA transcripts or deletion of FHIT. In 6 (16%) AML patients, 3 (19%) ALL patients, and 5 (15%) CML patients, the wild-type product was absent. Some patient's samples - 6 (42%) AML, 6 (38%) ALL, and 14 (41%) CML revealed aberrant FHIT transcripts in addition to a normal-sized band. Samples from healthy donors (PB, n = 12; BM, n = 5) did not indicate any abnormal expression. Eleven isolated fragments from various patterns of FHIT gene expression were investigated using cDNA sequencing. Sequence analysis revealed deletion of exon 4-8, exon 5-8, and exon 5-6 in various leukemias, as well as the deletion of the full FHIT gene sequence. The fused transcripts included: exon 3 and exon 9, exon 3 and exon 7, exon 4 and exon 9, exon 5 and exon 7. Sequence analysis of aberrant fragments present in samples from an AML and a CML patients was detected for point mutations and insert mutations located in exons 2, 8 and 10, plus a variety of aberrant transcripts. Deletion or aberrant FHIT mRNA transcripts in 50/98 (51%) leukemia patients were found. All samples with aberrant FHIT lacked gene product. A Kaplan-Meier plot of survival in patients with AML in relation to FHIT expression revealed that aberrance or loss of FHIT gene significantly correlated with a low clinical remission rate and poor overall survival.
Acid Anhydride Hydrolases ; genetics ; Base Sequence ; Gene Deletion ; Humans ; Leukemia ; genetics ; metabolism ; Lymphocytes ; metabolism ; Molecular Sequence Data ; Mutation ; Neoplasm Proteins ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; analysis

OBJECTIVETo investigate the effect of fragile histidine triad (FHIT) gene on the apoptosis of gastric cancer cells.

METHODSFHIT gene was transfected into gastric cancer cell line MGC-803 by liposome. Western blot and immunohistochemical assay were employed to detect the expression of FHIT. Apoptosis and cell cycle were analyzed by flow cytometry (FCM). Morphological changes were observed by light and electron microscope.

RESULTSStable expression of FHIT protein in transfected MGC-803 cells was obtained. The rates of G0/G1 phase and apoptosis cells were significantly higher in FHIT-transfected MGC-803 cells than those in control cells (64.4%compared with 49.5%and 32.0%compared with 12.2%, respectively).

CONCLUSIONThe results indicate that FHIT gene might be involved in the apoptosis of gastric cancer cells.

Acid Anhydride Hydrolases ; Apoptosis ; Cell Line, Tumor ; Flow Cytometry ; Genes, Tumor Suppressor ; Humans ; Immunohistochemistry ; Neoplasm Proteins ; analysis ; genetics ; Stomach Neoplasms ; genetics ; pathology ; prevention & control ; Transfection

OBJECTIVETo investigate the loss of heterozygosity (LOH) and microsatellite instability (MSI) of fragile histidine triad (FHIT) gene in laryngeal squamous cell carcinoma (LSCC).

METHODSUsing polymerase chain reaction-simple sequence length polymorphism (SSLP) -silver staining technique, 41 samples of LSCC were detected for LOH and MSI with microsatellite sites D3S1234 and D3S1300.

RESULTSLOH was found in 44.4% (16/36) and 36.4% (12/33) of informative cases at D3S1234 and D3S1300 respectively, while MSI at the two loci was found in 19.4% (7/36) and 24.2% (8/33) of informative cases respectively. In 41 cases of LSCC, 38 cases were informative at either locus. 52.6% (20/38) of cases had LOH while 28.9% (11/38) had MSI. The rate of LOH was related to the tumor TNM stage, pathological grade, lymph node metastasis and tumor relapse (P < 0.05). The rate of MSI was related to the tumor lymph node metastasis (P < 0.05 ).

CONCLUSIONThe frequencies of LOH and MSI in the two microsatellite sites, D3S1234 and D3S1300, of FHIT gene might play a role in carcinogenesis and development of LSCC and might provide some new approaches for early gene detection for LSCC.

Acid Anhydride Hydrolases ; genetics ; Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; pathology ; Female ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Loss of Heterozygosity ; Male ; Microsatellite Instability ; Middle Aged ; Neoplasm Proteins ; genetics

OBJECTIVETo study the relationship between aberrant FHIT transcripts and hepatocellular carcinoma (HCC).

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and single strand conformational polymorphism (SSCP) assays were used to analyze the transcripts and mutations of FHIT gene in 24 matched tumorous tissues and para-tumorous tissues from patients with HCC and in 4 normal liver tissues.

RESULTSAberrant FHIT transcripts were observed in 11 out of 24 (46%) tumorous tissues and in 2 (8%) of the matched para-tumorous tissues.

CONCLUSIONFHIT aberrant transcripts may play an important role in the pathogenesis of hepatocellular carcinoma.

Acid Anhydride Hydrolases ; Carcinoma, Hepatocellular ; genetics ; Humans ; Liver Neoplasms ; genetics ; Mutation ; Neoplasm Proteins ; genetics ; Polymorphism, Single-Stranded Conformational ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction