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Objective To investigate the association between fragile histidine triad (FHIT) gene methylation,abnormal protein expression and HPV16 infection as well as their interactions in cervical carcinogenesis.Methods A total of 108 patients with normal cervical (NC),142 cases of cervical intraepithelial neoplasia (CIN1,n=72;CIN2 +,n=70),and 100 new cases of cervical squamous cell carcinoma (SCC) were chosen from the Shanxi Tumor Hospital,Second Hospital of Shanxi Medical University,Maternal and Child Health Center in Taiyuan and Jiexiu during September 2009 and March 2011.HPV16 was detected by multiple PCR.FHIT methylation and protein expression levels were detected by methylation specific PCR (MSP) and Western Blot,respectively.All the data were performed with SPSS 20.0 statistical softvare.Differences among groups were assessed by Chi-square and Kruskal-Wallis tests.The interaction effects were evaluated by additive model.Results The prevalence rates of HPV16 infection in CIN1 (45.8％),CIN2+ (68.6％) and SCC (73.0％) were significantly higher than that in NC (28.7％,P＜0.001).In NC,CIN1,CIN2+ and SCC groups,the FHIT gene methylation rates were 3.7％,13.9％,21.4％ and 38.0％ while the protein expression levels were 1.255 ± 0.130,1.184 ± 0.172,1.133 ± 0.126 and 1.099 ± 0.148,respectively.Differences among the groups were statistical significant (P＜0.001).With increasing degrees of cervical lesions,the HPV16 infection rate (x2=47.623,P＜0.001),FHIT methylation rate (x2=40.147,P＜0.001) and the rate of FHIT protein low expression (x2=65.098,P＜0.001) were all gradually increasing.There appeared positive additive interaction between FHIT methylation,FHIT protein low expression and infection of HPV16.Conclusion Hypermethylation of FHIT gene,low expression of FHIT protein and HPVI6 infection could increase the risk of cervical cancer and precancerous lesions.These results suggested that there might be synergistic action between FHIT gene hypermethylation and HPV16 infection in the progression of cervical cancer and the same was true between the low expression of FHIT protein and HPV 16 infection.
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Objective To investigate the association between fragile histidine triad (FHIT) gene methylation,abnormal protein expression and HPV16 infection as well as their interactions in cervical carcinogenesis.Methods A total of 108 patients with normal cervical (NC),142 cases of cervical intraepithelial neoplasia (CIN1,n=72;CIN2 +,n=70),and 100 new cases of cervical squamous cell carcinoma (SCC) were chosen from the Shanxi Tumor Hospital,Second Hospital of Shanxi Medical University,Maternal and Child Health Center in Taiyuan and Jiexiu during September 2009 and March 2011.HPV16 was detected by multiple PCR.FHIT methylation and protein expression levels were detected by methylation specific PCR (MSP) and Western Blot,respectively.All the data were performed with SPSS 20.0 statistical softvare.Differences among groups were assessed by Chi-square and Kruskal-Wallis tests.The interaction effects were evaluated by additive model.Results The prevalence rates of HPV16 infection in CIN1 (45.8％),CIN2+ (68.6％) and SCC (73.0％) were significantly higher than that in NC (28.7％,P＜0.001).In NC,CIN1,CIN2+ and SCC groups,the FHIT gene methylation rates were 3.7％,13.9％,21.4％ and 38.0％ while the protein expression levels were 1.255 ± 0.130,1.184 ± 0.172,1.133 ± 0.126 and 1.099 ± 0.148,respectively.Differences among the groups were statistical significant (P＜0.001).With increasing degrees of cervical lesions,the HPV16 infection rate (x2=47.623,P＜0.001),FHIT methylation rate (x2=40.147,P＜0.001) and the rate of FHIT protein low expression (x2=65.098,P＜0.001) were all gradually increasing.There appeared positive additive interaction between FHIT methylation,FHIT protein low expression and infection of HPV16.Conclusion Hypermethylation of FHIT gene,low expression of FHIT protein and HPVI6 infection could increase the risk of cervical cancer and precancerous lesions.These results suggested that there might be synergistic action between FHIT gene hypermethylation and HPV16 infection in the progression of cervical cancer and the same was true between the low expression of FHIT protein and HPV 16 infection.
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Objective To study fragile histidine triad gene(FHIT) gene mRNA expression in 60 samples of papillary thyroid carcinoma(PTC) and matched adjacent non-concerous tissues (NCE),and to explore the role of FHIT gene in the development in PTC.Methods Reverse transcription polymerase chain reaction(RT-PCR) was used to detect FHIT gene mRNA expression in 60 samples of PTC and matched NCE.Results In PTC tissues,the positive rate of FHIT gene mRNA expression was 45.0％ (27/60),lower than that in NCE tissues (100.0％,60/60).FHIT gene mRNA expression was correlated with pathology stage,TNM stage and lymph node metastasis (P ＜ 0.05).Conclusion FHIT loss and transcript abnormality might play important roles in proliferation and metastasis of PTC,and might be a useful marker for evaluating the biological behavior of PTC.
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Objective 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. Methods 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. Results 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 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/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. Conclusion 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.
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Objective 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. Methods 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. Results 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 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/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. Conclusion 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.
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Fragile histidine triad(FHIT) gene is a tumor suppressor gene that locates on chromosome 3p14.2.FHIT can induce cell apoptosis and inhibit cell growth by activating caspase,inhibiting PI3K-Akt-survivin signal pathway and phosphorylation of I?B-?,and binding with microtube.The inactivation of FHIT is closely related with carcinogenesis.The advances in research on the structure,biological function,relationship between inactivation and carcinogenesis,and gene therapy of FHIT are reviewed in this paper.
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Objective To investigate the effects of exogenous fragile histidine triad(FHIT) gene on apoptosis of human glioma cell line U87. Methods By the method of liposome transfection,plasmids pcDNA3.1/myc-His(-)B-FHIT and pcDNA3.1/myc-His(-)B were transfected into glioma cell line U87.U87 cells were divided into three groups: U87-FHIT group,U87 cells transfected by plasmids pcDNA3.1/myc-His(-)B-FHIT;U87-vector group,U87 cells transfected by plasmids pcDNA3.1/myc-His(-)B;and blank control group,U87 cells without transfection.The expression of exogenous FHIT protein was detected by Western blot and immunofluorescence staining.The effects of FHIT on the growth characteristics of U87 were observed by MTT and flow cytometry. Results Growth inhibitory rate and apoptosis rate of the cells in U87-FHIT group were significantly higher than those in U87-vector group and blank control group(P
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Objective To investigate the clinical significance of the expression of fragile histidine triad (FHIT) protein in primary hepatocellular carcinoma (HCC).Methods Immunohistochemical (S-P) method was used to detect the expression of FHIT in the 46 patients with HCC and 10 normal controls.Results In the patients with HCC,the expression rate of FHIT in the tumor tissue was 56.52%,significantly lowerthan that in adjacent non-tumor tissue and normal tissue.The absence of FHIT protein was correlated neither with the size,tumor capsule,serum AFP level nor with chronic hepatitis B virus infection and cirrhosis of liver.There was significant relationship between the expression of FHIT and differentiation and thrombus in the portal vein.Conclusion The loss of FHIT gene protein is a frequent event in HCC.Furthermore,the loss is closoly related to tumor prognosis prognosis and FHIT loss.
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Objective:To study dynamic changes of fragile histidine triad(FHIT) gene expression in ionizing radiation injury and radiation carcinogenesis. Methods: BEAS-2B cells were divided into 0.5,1,2,4,8,16 Gy irradiation groups and control group. In 24 h, 72 h and 10 d after irradiation, the expression of FHIT gene was studied with single-cell RT-PCR and DNA sequencing separately. Results: Different types of FHIT gene mutations occured in different phases after irradiation with different doses (All mutations were exon deletion mutations by DNA sequencing), while abnormal FHIT gene was not detected in control group. The percentage of mutation in 0.5,1,2,4,8,16 Gy dose groups was 52.6%,66.7%,57.9%,76.5%,64.7% and 81.3% respectively 24 h after irradiation;17.6%,22.2%,50.0%,47.4%,47.1% and 68.4% respectively 72 h after irradiation;and 21.1%,25.0%,60.0%,57.9%,61.1% and 68.4% respectively 10 d after irradiation. Conclusion: These results suggest that ionizing radiation can cause deletion of FHIT gene.