ABSTRACT
<p><b>OBJECTIVE</b>To correlate the clinical characteristics with mutations of the STK11 and FHIT genes in 16 patients with Peutz-Jeghers syndrome (PJS).</p><p><b>METHODS</b>Potential mutations in the coding regions and flanking sequences of the STK11 and FHIT genes were detected with PCR and Sanger sequencing.</p><p><b>RESULTS</b>Of 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%.</p><p><b>CONCLUSION</b>Mutations 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.</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Acid Anhydride Hydrolases , Genetics , Base Sequence , DNA Mutational Analysis , Exons , Molecular Sequence Data , Mutation , Neoplasm Proteins , Genetics , Pedigree , Peutz-Jeghers Syndrome , Genetics , Protein Serine-Threonine Kinases , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the interaction between folate deficiency and aberrant expression related to fragile histidine triad (FHIT) gene in the progression of cervical cancerization.</p><p><b>METHODS</b>A total number of 80 patients with histological diagnosis of cervix inflammation (CI), 55 cervical intraepithelial neoplasm I (CIN I), 55 cervical intraepithelial neoplasm II/III (CIN II/III) and 64 cervical squamous cell carcinoma (SCC) were included in this study. Levels of serum folate were detected by microbiological assay method and the methylation status of FHIT gene CpG islands was tested by methylation-specific PCR (MSP). FHIT protein levels were measured by Western blot. In vitro, cervical cancer cell lines CaSki (HPV16-positive) was treated with different concentrations of folate. Proliferation and apoptosis of cells, methylation of FHIT gene and the levels of FHIT protein expression were measured in each group. All analyses were performed with SPSS (version 17.0) statistical software. Differences among groups were assessed by chi-square test, Kruskal-Wallis test. Spearman correlation, and the interaction effects were evaluated by additive model.</p><p><b>RESULTS</b>The levels of serum folate (H = 59.08, P < 0.001) and FHIT protein expression (H = 50.93, P < 0.001) decreased gradually along with the severity of cervix lesions, while the methylation rates of FHIT gene CpG islands increased (trend χ² = 28.34, P < 0.001). Both levels of serum folate levels and FHIT protein expression were positively correlated (r = 0.213, P = 0.001), with an additive interaction seen between them in CIN I, CIN II/III, SCC groups. In vitro, both rates related to proliferation inhibition (r = 0.98, P < 0.001) and apoptosis (r = 0.99, P < 0.001) together with the levels of FHIT protein expression (r = 0.97, P < 0.001) were all increased gradually with the increase of folate concentration while the methylation status of FHIT gene CpG islands all changed from positive to negative gradually.</p><p><b>CONCLUSION</b>Results from our study revealed that both folate deficiency and FHIT protein aberrant low expression might increase the risk of developing cervical cancer and cervix precancerous lesions, and thus play a synergistic action in the progression of cervical cancerization.</p>
Subject(s)
Female , Humans , Acid Anhydride Hydrolases , Metabolism , Apoptosis , Carcinoma, Squamous Cell , Pathology , Uterine Cervical Dysplasia , Pathology , Disease Progression , Folic Acid , Blood , Folic Acid Deficiency , Epidemiology , Human papillomavirus 16 , Neoplasm Proteins , Metabolism , Polymerase Chain Reaction , Uterine Cervical Neoplasms , PathologyABSTRACT
<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>180 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.</p><p><b>RESULTS</b>Relative 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).</p><p><b>CONCLUSION</b>Occupational 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.</p>
Subject(s)
Humans , Acid Anhydride Hydrolases , Genetics , Coal Tar , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , DNA Methylation , Leukocytes , Neoplasm Proteins , Genetics , Occupational Exposure , Promoter Regions, Genetic , Telomere , Tumor Suppressor Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Cervical 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.</p><p><b>RESULTS</b>Folate 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.</p><p><b>CONCLUSION</b>Our 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.</p>
Subject(s)
Female , Humans , Acid Anhydride Hydrolases , Genetics , Metabolism , Apoptosis , Cell Line, Tumor , Cell Proliferation , Culture Media , Chemistry , DNA Methylation , Folic Acid , Pharmacology , Neoplasm Proteins , Genetics , Metabolism , Uterine Cervical Neoplasms , Genetics , PathologyABSTRACT
<p><b>BACKGROUND</b>It 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.</p><p><b>METHODS</b>Data 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.</p><p><b>RESULTS</b>Complete 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%.</p><p><b>CONCLUSIONS</b>These 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.</p>
Subject(s)
Humans , Acid Anhydride Hydrolases , Metabolism , Galectin 3 , Metabolism , Immunohistochemistry , Ki-67 Antigen , Metabolism , Neoplasm Proteins , Metabolism , Parathyroid Neoplasms , Metabolism , Tumor Suppressor Proteins , MetabolismABSTRACT
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.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Acid Anhydride Hydrolases , Genetics , Azacitidine , Therapeutic Uses , DNA , Blood , DNA Methylation , Myelodysplastic Syndromes , Blood , Drug Therapy , Neoplasm Proteins , Genetics , Promoter Regions, GeneticABSTRACT
En Chile, el cáncer es la segunda causa de muerte después de las enfermedades cardiovasculares. Los principales cánceres asociados a muerte en mujeres fueron mama, estómago, vesícula biliar, broncopulmonar y cérvico uterino. Alteraciones de las E-cadherinas han sido relacionadas con varios tipos de cáncer, ya que uno de los principales eventos involucrados con su disfunción es el gatillar la invasión y metástasis del tumor. La inactivación del gen CDH1 ha sido demostrada en el cáncer gástrico difuso y el cáncer de mama lobulillar. Asimismo, la inactivación del gen FHIT parece estar asociado con la progresión a neoplasias más agresivas. Se realizaron determinaciones inmunohistoquímicas (IHQ) en fibroadenomas mamarios y cánceres previamente diagnosticados por RE, RPg y Her2, mostrando positividad en todos los casos. La detección (IHQ) de la expresión de FHIT y E-cadherina en tejidos con patologías benignas y malignizados, puede aportar una importante información diagnóstica y pronóstica en el cáncer de mama.
In Chile, cancer is the second leading cause of death after cardiovascular diseases. The major death-related cancers in women were breast, stomach, gallbladder, lung and cervical cancer. Alterations of E-cadherin have been linked to various cancers, as one of the main events involved in its dysfunction is the trigger of tumor invasion and metastasis. CDH1 gene inactivation has been demonstrated in diffuse gastric cancer and lobular breast cancer. Furthermore, inactivation of the FHIT gene to be associated with progression to more aggressive tumors. Immunohistochemistry (IHC) determinations were performed in fibroadenomas and breast cancers previously diagnosed by ER, PgR and Her2, showing positivity in all cases. Immunohistochemical detection of FHIT and E-cadherin expression in tissues with benign disease and malignant, may provide an important diagnostic and prognostic information in breast cancer.
Subject(s)
Humans , Female , Acid Anhydride Hydrolases/metabolism , Cadherins/metabolism , Breast Neoplasms/diagnosis , Breast Neoplasms/metabolism , Neoplasm Proteins/metabolism , Carcinoma, Ductal, Breast/diagnosis , Carcinoma, Ductal, Breast/metabolism , Carcinoma, Lobular/diagnosis , Carcinoma, Lobular/metabolism , Fibroadenoma/diagnosis , Fibroadenoma/metabolism , ImmunohistochemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the expressions of the FHIT and PTEN genes and their significance in prostate cancer.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSION</b>FHIT 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.</p>
Subject(s)
Aged , Humans , Male , Middle Aged , Acid Anhydride Hydrolases , Metabolism , Adenocarcinoma , Metabolism , Pathology , Immunohistochemistry , Neoplasm Proteins , Metabolism , PTEN Phosphohydrolase , Metabolism , Prostatic Neoplasms , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To study the relationship between FHIT and WWOX expression and clinicopathological features in hepatocellular carcinoma (HCC).</p><p><b>METHOD</b>The expression of FHIT and WWOX were determined by immunohistochemistry in 142 patients with HCC.</p><p><b>RESULTS</b>Absent 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).</p><p><b>CONCLUSIONS</b>Absent/reduced FHIT and WWOX expression is associated with poor prognosis.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Acid Anhydride Hydrolases , Genetics , Carcinoma, Hepatocellular , Genetics , Pathology , Gene Expression Profiling , Liver Neoplasms , Genetics , Pathology , Neoplasm Proteins , Genetics , Oxidoreductases , Genetics , Prognosis , Tumor Suppressor Proteins , Genetics , WW Domain-Containing OxidoreductaseABSTRACT
<p><b>OBJECTIVE</b>To study the expression and significance of fragile histidine triad (FHIT) and Ki-67 in transformed epithelial cells induced by Yunnan tin mine dust.</p><p><b>METHODS</b>Every 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.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSIONS</b>FHIT 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.</p>
Subject(s)
Humans , Acid Anhydride Hydrolases , Metabolism , Cell Line , Cell Transdifferentiation , China , Dust , Epithelial Cells , Cell Biology , Metabolism , Ki-67 Antigen , Metabolism , Lung , Cell Biology , Neoplasm Proteins , Metabolism , Tin , ToxicityABSTRACT
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.
Subject(s)
Female , Humans , Male , Acid Anhydride Hydrolases , Genetics , Metabolism , Immunohistochemistry , Mouth Mucosa , Metabolism , Mouth Neoplasms , Metabolism , Neoplasm Proteins , Genetics , Metabolism , Oral Submucous Fibrosis , Metabolism , Proto-Oncogene Proteins c-mdm2 , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of fragile histidine triad (FHIT) gene transfection on human colorectal cancer cell line SW480 through up-regulation of caspase-8 expression.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>At 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.</p><p><b>CONCLUSIONS</b>FHIT 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.</p>
Subject(s)
Humans , Acid Anhydride Hydrolases , Genetics , Apoptosis , Caspase 8 , Metabolism , Cell Line, Tumor , Cell Proliferation , Colonic Neoplasms , Genetics , Pathology , Neoplasm Proteins , Genetics , RNA, Messenger , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of fragile histidine triad in endometriosis and investigate the pathogenesis of endometriosis.</p><p><b>METHODS</b>immunohistochemistry 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.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSIONS</b>Fhit may play an important role in the development of endometriosis.</p>
Subject(s)
Adult , Female , Humans , Middle Aged , Acid Anhydride Hydrolases , Metabolism , Endometriosis , Metabolism , Pathology , Endometrium , Metabolism , Neoplasm Proteins , MetabolismABSTRACT
OBJECTIVE@#To analyze the mutation and abnormal expression of the FHIT gene in human hypopharyngeal carcinoma.@*METHOD@#Matched normal and cancerous tissues from 24 patients with hypopharyngeal squamous cell carcinoma were obtained immediately after surgery. Total RNA was extracted, the FHIT gene was detected by nested RT-PCR and DNA sequencing technology.@*RESULT@#Normal sized FHIT transcript was detected in 23 of the 24 cases of normal matched tissues. Aberrant FHIT transcripts were found in 9/24 (37.5%) cases in hypopharyngeal carcinoma. Aberrant FHIT transcripts rate of well-differentiated, moderately differentiated and poorly differentiated squamous cell carcinoma, was 28.6% (2/7), 50.0% (4/8) and 33.3% (3/9), respectively. There the carcinoma with FHIT aberrant transcripts was neither corresponding to histological grade (P>0.05) nor to lymphatic metastasis. The sequence analyses of the two aberrant cDNAs revealed absence of exon 8 and exon 7-9. All initial deletion were in conjunction of exons.@*CONCLUSION@#High deletion rate of the FHIT gene in Chinese hypopharyngeal squamous cell carcinoma suggested the FHIT gene, a candidate tumor suppressor gene at 3p14.2, plays an important role in the tumor carcinogenesis, development and progression of the tumor, and thus may become a new prognostic marker in hypopharyngeal carcinoma.
Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Acid Anhydride Hydrolases , Genetics , Base Sequence , Carcinoma, Squamous Cell , Genetics , Exons , Gene Deletion , Hypopharyngeal Neoplasms , Genetics , Molecular Sequence Data , Mutation , Neoplasm Proteins , GeneticsABSTRACT
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.
Subject(s)
Animals , Female , Humans , Mice , Acid Anhydride Hydrolases/genetics , Cell Line, Tumor , Cholangiocarcinoma/genetics , Gene Expression Profiling , Mice, Inbred BALB C , Mutation , Neoplasm Proteins/genetics , Oligonucleotide Array Sequence Analysis , Sarcoma/genetics , Tumor Suppressor Protein p53/geneticsABSTRACT
Background: The loss of tumor suppresor gene function damages the defensive mechanisms that protect the indemnity of genetic material. Promoter gene methylation is one of the inactivation mechanisms of suppressor genes. Aim: To study the methylation pattern of a group of genes in biopsy samples of gastrointestinal tumors. Material and methods: Forty eight gastric, 25 gallbladder, 24 colon and 6 pancreas cancer biopsy samples were randomly selected. The methylation pattern of CDH1, FHIT, CDKN2A, APC and MLH1 genes, was studied using a specific polymerase chain reaction test for methylation. Demographic, morphological and follow up variables of patients bearing the tumors were also analyzed. Results: The general methylation frequency of CDH1, FHIT, CDKN2A, APC and MLH1 genes was 64.1, 56, 39.8, 18.1 and 34 percent respectively. In gastric cancer samples there was a correlation between APC gene methylation and well differentiated tumors; between CDH1 methylation and Lauren diffuse type and the presence of three or more metastasic lymph nodes; between FHIT, CDKN2A and CDH1 gene methylation and male gender. In ¡ess differentiated gallbladder tumors, the frequency of CDH1 methylation was higher. There was a tendency towards a lower survival in colon and gastric cancer when MLH1 (p =0.07) y CDKN2A (p= 0.06) were methylated, respectively. Conclusions: An abnormal methylation pattern was associated with morphological features in gastric and gallbladder cancer and with a tendency towards a lower survival in colon and gastric cancer.
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Carcinoma/genetics , DNA Methylation/genetics , Gallbladder Neoplasms/genetics , Gastrointestinal Neoplasms/genetics , Pancreatic Neoplasms/genetics , Kaplan-Meier Estimate , Acid Anhydride Hydrolases/genetics , Acid Anhydride Hydrolases/metabolism , Cadherins/genetics , Carcinoma/metabolism , Gallbladder Neoplasms/metabolism , Gastrointestinal Neoplasms/metabolism , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Nucleic Acid Amplification Techniques , Pancreatic Neoplasms/metabolism , Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To study the methylation status of fragile histidine triad (FHIT) gene promoter in patients with myelodysplastic syndrome (MDS) and its clinical relevance.</p><p><b>METHODS</b>Methylation-specific PCR (MSP) was used to detect FHIT promoter methylation in bone marrow samples from 54 MDS cases.</p><p><b>RESULTS</b>Hypermethylation of FHIT promoter was detected in 26 cases (48.1%). Association was not found between FHIT gene hypermethylation and sex, hematologic parameters and chromosomal abnormalities of MDS patients, but found between FHIT gene hypermethylation and age of the MDS cases. Although significant difference was not observed in the frequencies of FHIT gene hypermethylation among patients with refractory anemia/refractory anemia with ringed sideroblasts (RA/RAS) (1/6, 16.7%), refractory anemia/refractory anemia with ringed sideroblasts (RCMD) and refractory cytopenia with multilineage dysplasia with ringed blasts (RCMD-RS) (6/19, 31.6%), refractory anemia with excess blasts-1 (RAEB-1) (7/11, 63.6%), refractory anemia with excess blasts-2 (RAEB-2) (4/7, 57.1%) and refractory anemia with excess blasts in transformation/acute myeloid leukemia (RAEBt/AML) (8/11, 72.7%)(chi-square=8.417, P=0.077), it was observed in patients in early stages (RA/RAS and RCMD) (7/25, 28.0%), advanced stages (RAEB-1 and RAEB-2)(11/18, 61.1%) and RAEBt/AML (8/11, 72.7%) (chi-square=7.938, P=0.019). Furthermore, there was a positive correlation between the frequency of FHIT gene hypermethylation and different IPSS groups (chi-square=10.110, P=0.018).</p><p><b>CONCLUSION</b>FHIT gene hypermethylation might be one of the molecular events involved in the disease progression of MDS.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Acid Anhydride Hydrolases , Genetics , Age Factors , Base Sequence , DNA Methylation , Molecular Sequence Data , Myelodysplastic Syndromes , Classification , Genetics , Pathology , Neoplasm Proteins , Genetics , Polymerase Chain Reaction , Promoter Regions, Genetic , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the suppression effect of exogenous fragile histidine triad (FHIT) gene on biological property of MEC-1 cells.</p><p><b>METHODS</b>In 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.</p><p><b>RESULTS</b>It 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.</p><p><b>CONCLUSION</b>Exogenous FHIT gene could suppress the proliferation, tumorigenicity and improve the differentiation of MEC-1 cells, in vitro and in vivo.</p>
Subject(s)
Animals , Humans , Mice , Acid Anhydride Hydrolases , Apoptosis , Carcinoma, Mucoepidermoid , Cell Cycle , Cell Line, Tumor , Histidine , Mice, Nude , Neoplasm Proteins , TransfectionABSTRACT
<p><b>BACKGROUND</b>WWOX and FHIT are two candidate tumor suppressor genes located in active fragile sites, the damage of which has been associated with the development of breast cancer. The association of the expression of these genes and the development of breast cancer has not been fully explored. We evaluated mRNA and protein expression of WWOX and FHIT in breast tissue with normal histological appearances, atypical ductal hyperplasia, ductal carcinoma in situ, and invasive cancer to see if a progressive decline in expression was present.</p><p><b>METHODS</b>Reverse transcription-polymerase chain reaction and Western blotting were used to evaluate the specimens for mRNA and protein expression, including 28 specimens with normal tissue, 28 specimens with atypical ductal hyperplasia, 33 specimens with ductal carcinoma in situ, and 51 specimens with invasive ductal carcinoma.</p><p><b>RESULTS</b>Compared with in situ and invasive cancer specimens, both normal and atypical hyperplasia specimens had greater rates of detectable mRNA (WWOX rate ratio = 2.95, 95% CI 1.24 - 7.08; FHIT rate ratio = 4.58, 95% CI 1.82 - 11.81) and Western blotting detectable protein (WWOX rate ratio = 4.12, 95% CI 1.63 - 10.73; FHIT rate ratio = 3.76, 95% CI 1.44 - 10.06). For both proteins, differences between normal and atypical hyperplasia specimens and between in situ and invasive carcinoma specimens were explainable by chance (P > 0.05 for each analysis). Within each histological category, differences among fractions of specimens showed that FHIT and WWOX mRNA and protein expression were explainable by chance (P > 0.05 for each analysis).</p><p><b>CONCLUSION</b>Expression of FHIT and WWOX decreases along with breast tissue progress from a normal histological appearance to atypical ductal hyperplasia, in situ cancer, and the final invasive cancer.</p>
Subject(s)
Female , Humans , Acid Anhydride Hydrolases , Genetics , Breast , Pathology , Breast Neoplasms , Genetics , Chromosome Fragile Sites , Genes, Tumor Suppressor , Hyperplasia , Neoplasm Proteins , Genetics , Oxidoreductases , Genetics , Tumor Suppressor Proteins , Genetics , WW Domain-Containing OxidoreductaseABSTRACT
OBJECTIVE@#To investigate the role and clinical significance of the expression of FHIT gene in development and progression of laryngeal squamous cell carcinoma(LSCC).@*METHOD@#The expression FHIT protein was detected by immunohistochemistry method in the 52 cases with LSCC and 23 cases of adjacent cancer tissues and 10 cases of laryngeal normal mucosa. The expression of FHIT was analyzed in LSCC with different clinicopathological parameters.@*RESULT@#The expression of FHIT in normal tissues (90.0%) and adjacent tissues (78.3%) was obvious higher than that in LSCC (46.2%), which had statistical significant difference (P < 0.01). There was a positive correlation between FHIT expression and TNM staging, lymph node metastasis; but not pathological grade.@*CONCLUSION@#The loss of FHIT expression may be correlated with the development and progression of laryngeal carcinoma. FHIT may be an important tumor suppressor gene in LSCC.