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Abstract Apurinic/apyrimidinic endonuclease 1 (APE1) is a multifunctional key protein. Recent studies suggest APE1 is closely associated with inflammatory response, but its role in asthma remains unknown. We recruited 116 patients with asthma, including 50 with severe asthma (NSA) and 66 with non-severe asthma (SA), and 140 controls. Serum APE1 was detected using the ELISA method. APE1 mRNA in peripheral blood neutrophils and eosinophils were detected using real-time PCR assays. Compared to healthy controls, we observed significant elevations of serum APE1 mRNA levels in peripheral neutrophils (~1.75 folds increase, p<0.05) and eosinophils (~2.2 folds increase, p<0.05) in patients with asthma. The peripheral blood neutrophil APE1 mRNA can distinguish asthmatic patients from healthy controls with the area under the curve (AUC) 0.893 and a 95% confidence interval (CI) 0.847-0.938 (p < 0.001). Also the APE1 mRNA can identify severe asthma from non-severe asthma (AUC 0.759, 95% CI, 0.674-0.846; p < 0.001). However, The serum APE1 and eosinophil mRNA levels did not correlate with asthma incidence and severity. Our finding confirms the association between APE1 and asthma and suggests that peripheral blood neutrophil APE1 mRNA may be used as a marker for this condition.
Resumen La endonucleasa apurínica/apirimidínica 1 (APE1) es una proteína clave multifuncional. Estudios recientes sugieren que APE1 está estrechamente asociada con la respuesta inflamatoria, pero hasta el momento se desconoce su papel en el asma. Reclutamos a 116 pacientes con asma, incluidos 50 con asma grave (NSA) y 66 con asma no grave (SA), y 140 controles. Se detectó APE1 en suero usando el método ELISA. El ARNm de APE1 en neutrófilos y eosinófilos de sangre periférica se detectó mediante ensayos de PCR en tiempo real. En comparación con los controles sanos, observamos una elevación significativa de los niveles séricos de ARNm de APE1 en pacientes con asma en neutrófilos periféricos (aumento de ~1,75 veces, p<0,05) y eosinófilos (aumento de ~2,2 veces, p<0,05). El ARNm de APE1 de neutrófilos de sangre periférica puede distinguir a los pacientes asmáticos de los controles sanos con un área bajo la curva (AUC) de 0,893 y un intervalo de confianza (IC) del 95% de 0,847 a 0,938 (p<0,001). Además, el ARNm de APE1 puede identificar el asma grave del asma no grave (AUC 0,759, IC del 95%, 0,674-0,846; p < 0,001). Sin embargo, el nivel sérico de APE1 y ARNm de eosinófilos no mostró correlación con la incidencia y la gravedad del asma. Nuestro hallazgo confirma la asociación entre APE1 y asma y sugiere que el ARNm de APE1 de neutrófilos en sangre periférica puede usarse como marcador para el asma.
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Objective To examine the single nucleotide polymorphism(SNP) of apurinic/apyrimidinic endonuclease1 (APE1) in primary small cell carcinoma of esophagus(PSEC) ,then investigate the relationship between these SNPs and the prognosis.Methods Sixty cases first-treated patients with PSEC were recruited, patients with esophageal squamous cell carcinoma (ESCC) and healthy blood donors were recruited as positive and negative controls.APE1 (Asp148Glu) of the patients with PSEC and controls were genotyped by the TaqMan method.Every patient was treated with platinum-based chemotherapy(EP regimen for PSEC and TP regimen for ESCC)and radiotherapy(3D-CRT) ,then every case was followed-up for 2 years.The relationship between these SNPs and the follow-up outcome was analyzed.Results Compared with the ESCC group and control group, APE1 148 pure mutant(Glu/Glu) of PSEC group increased significantly(PSEC group was 40％ (12/30), ESCC group was 13.3％ (4/30) , control group was 10％ (2/20)), the difference was statistically significant (x2 =7.248,P =0.027).According to data of following-up, there was a significant increase in rate of progress (1year:40.0％ (12/30) vs 16.7％ (5/30), x2 =4.022, P =0.045;2 years: 86.7％ (26/30) vs 40.0％ (12/30) ,P =0.004) and a significant decrease in survival (33.3％ (10/30) vs 76.7％ (23/30)) of PSEC compared with ESCC.The SNPs of APE1 Asp148Glu was significantly correlated with frequency of progress, a significant increase was found in rate of progress of the patients with mutant type(Asp/Glu±Glu/Glu) compared with wild genotype(1 year: 50.0％(11/22) ,x2 =3.854,P=0.05;2 years: 81.8％ (19/22) ,x2 =10.519,P =0.001) ,the survival of the patients with mutant genotype was significantly lower than wild type (22.7％ (5/22) ,x2=10.77,P=0.001).Conclusion The most of polymorphisms of APE1(Asp148Glu) are mutation type in PSEC.Pure mutant genotype (APE1 148Glu/Glu) carry significant enhancement of progression.The polymorphisms of APE1 (Asp148Glu) maybe one of those molecular mechanisms of high frequency of progress and poor prognosis in PSEC.
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PURPOSE: Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein involved in DNA repair and redox modulation. Recently, serum and urinary APE1/Ref-1 levels were reported to be increased in patients with bladder cancer. Genetic variations of APE/Ref-1 are associated with the risk of cancer. However, the effect of APE1/Ref-1 variants on its secretory activity is yet unknown. METHODS: APE1/Ref-1 variants were evaluated by DNA sequencing analysis of reverse transcription polymerase chain reaction products in coding DNA sequences (CDS) of APE1/Ref-1 in bladder tissue samples from patients with bladder cancer (n=10). Secretory activity of APE1/Ref-1 variants was evaluated with immunoblot and enzyme-linked immunosorbent assay of the culture medium supernatants. RESULTS: Four different substitution mutants (D148E, I64V/D148E, W67R/D148E, and E86G/D148E) of APE1/Ref-1 were identified in bladder cancer specimens. However, deletion mutants of APE1/Ref-1 CDS were not found. The secretory activity of the APE1/Ref-1 variants (D148E, I64V/D148E, and E86G/D148E) was increased compared to that of wild type APE1/Ref-1. Furthermore, the secretory activity in basal or hyperacetylated conditions was much higher than that in APE1/Ref-1 D148E-transfected HEK293 cells. CONCLUSIONS: Taken together, our data suggest that the increased secretory activity of D148E might contribute to increased serum levels of APE1/Ref-1 in patients with bladder cancer.
Subject(s)
Humans , Base Sequence , Clinical Coding , DNA Repair , Enzyme-Linked Immunosorbent Assay , Genetic Variation , HEK293 Cells , Oxidation-Reduction , Point Mutation , Polymerase Chain Reaction , Reverse Transcription , Sequence Analysis, DNA , Urinary Bladder Neoplasms , Urinary BladderABSTRACT
PURPOSE: Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein that shows elevated expression in a number of cancers. We attempted to determine whether serum APE1/Ref-1 is elevated in patients with bladder cancer. MATERIALS AND METHODS: Serum APE1/Ref-1 levels were determined using enzyme-linked immunosorbent assay in serum from patients with bladder cancer who had not received chemotherapy or radiotherapy (n=51) and non-tumor controls (n=55). The area under the receiver operating characteristic area under the curve was applied to determine the correlation between clinical factors and the serum levels of APE1/Ref-1. RESULTS: Serum levels of APE1/Ref-1 in bladder cancer patients were significantly elevated compared to those of the control group (3.548+/-0.333 ng/100 muL [n=51] for bladder cancer vs. 1.547+/-0.319 ng/100 muL [n=55] for the control group), with a sensitivity and specificity of 93% and 59%, respectively. Serum APE1/Ref-1 levels are associated with tumor stage, grade, muscle invasion, and recurrence. CONCLUSION: Serum APE1/Ref-1 might be useful as a potential serologic biomarker for bladder cancer.
Subject(s)
Humans , Biomarkers , Drug Therapy , Enzyme-Linked Immunosorbent Assay , Radiotherapy , Recurrence , ROC Curve , Sensitivity and Specificity , Urinary Bladder Neoplasms , Urinary BladderABSTRACT
Objective To investigate the effect of APE1 on differentiation of peripheral blood mononuclear cells into osteoclast-like cells(OCL) which induced by macrophage colony stimulating factor (RANKL) and macrophage colony stimulating factor (M-CSF) .Methods Human peripheral blood mononuclear cells (PBMCs) were collected by density gradient separation ;Constructed APE1 siRNA expression vector Ad5v-APE1 siRNA was used to transfect PBMCs .Tartrate-resistant acid phosphatase (TRAP) method was conducted to identify the cells ,the expression level of APE1 was detected by Western blot ,the mRNA expression levels of Cathepsin K(CK) and V-ATPase were detected by RT-PCR .Results PBMCs transfected with APE1 siRNA had significantly lower protein expression of APE1 than untransfected cells (P< 0 .05) ;PBMCs could differentiate into OCL under the stimulation of RANKL and M-CSF ,the mRNA expression levels of CK and V-ATPase increased ;After APE1 siRNA treatment ,the number of OCL was reduced and the levels of CK and V-ATPase mRNA decreased .Conclusion PBMCs can differentiate into a large number of OCL induced by RANKL and M-CSF ,APE1 siRNA significantly inhibited differentiation of PBMCs into osteoclast-like cells , APE1 may be involved in the regulation of osteoclast-like differentiation process .
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Objective To detect the expression of apurinic/apyrimidinic endonuclease 1 (APEI) and explore its correlation with the expression of mutant p53 in hepatocellular carcinoma (HCC). Methods The expression of APE1 and mutant p53 was detected by SP immunohistochemical method in 10 specimens of normal liver tissue, 40 specimens of liver cirrhosis tissue and 103 specimens of HCC tissue which were collected at the Department of Pathology of Daping Hospital from 1991 to 2004. All data were analyzed by chi-square test, correla-tion analysis and K Independent-Samples Tests. Results The expression rate of APE1 in HCC was 100.0%, which was significantly higher than that in normal liver tissue (40.0%) and liver cirrhosis tissue (82.5%) (χ~2= 47.852, P < 0.01). The expression of APE1 was only detected in the nucleus in normal liver tissue. Ectopic expression of APE1 in cytoplasm was detected in liver cirrhosis tissue and HCC tissue, with the rate of 20.0% and 53.4%, respectively (χ~2=20.757, P <0.01). There was statistical difference in clinical staging and pathological grading of HCC with different combinations of APE1 expression (intranuclear or ectopic expression) and mutant p53 expression (positive or negative expression) (χ~2=12.910, 14.481, P < 0.01), and HCC with ectopic expression of APE1 and positive expression of p53 had high malignant degree. Conclusion Overexpression and ectopic expression of APE1 in cytoplasm may play important roles in the genesis and progression of HCC, and the ectopic expression of APE1 and p53 mutation may have synergistic effect.
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The role of apurinic/apyrimidinic endonuclease1/redox factor-1 (Ref-1) on the lead (Pb)-induced cellular response was investigated in the cultured endothelial cells. Pb caused progressive cellular death in endothelial cells, which occurred in a concentration- and time-dependent manner. However, Ref-1 overexpression with AdRef-1 significantly inhibited Pb-induced cell death in the endothelial cells. Also the overexpression of Ref-1 significantly suppressed Pb-induced superoxide and hydrogen peroxide elevation in the endothelial cells. Pb exposure induced the downregulation of catalase, it was inhibited by the Ref-1 overexpression in the endothelial cells. Taken together, our data suggests that the overexpression of Ref-1 inhibited Pb-induced cell death via the upregulation of catalase in the cultured endothelial cells.
Subject(s)
Catalase , Cell Death , Down-Regulation , Endothelial Cells , Hydrogen Peroxide , Superoxides , Up-RegulationABSTRACT
PURPOSE: The purpose of this study was to verify that the expressions of angiogenin, transforming growth factor-beta(TGF-beta), vascular endothelial growth factor(VEGF), human apurinic/apyrimidinic endonuclease(APEX) and tumor necrosis factoralpha(TNF-alpha) were associated with the tumorigenesis of the oral squamous cell carcinoma(OSCC). MATERIALS AND METHODS: Fifty-one samples of OSCC and fifteen normal oral mucosae were obtained to analyze the expression levels of above five factors. mRNA expressions were quantified by the quantitative competitive PCR(QC-PCR) method. After 2% agarose gel electrophoresis stained with ethidium bromide, the concentration of mRNA was calculated by a digital image analysis system. The expression levels of angiogenin, TGF-beta, VEGF, APEX and TNF-alpha were compared by unpaired Student's ttests between cancer and normal tissues. We analyzed statistically to find the cut-off values that would be useful as diagnostic markers, and the linear regression analysis between every two factors of these five factors by SAS system. RESULTS: All of these five factors (angiogenin: P<0.0037, TGF-beta: P<0.0001, VEGF: P<0.0102, APEX: P<0.0023, TNF-alpha: P<0.0074) were significantly correlated with OSCC. In the analysis to find the cut-off values for the diagnosis, we could not find any value that had a reasonable sensitivity and specificity. In the linear regression analysis, there were correlations between angiogenin and TNF-alpha, TGF-beta and VEGF, TGF-beta and APEX, TGF-beta and TNF-alpha, VEGF and APEX, VEGF and TNF-alpha, APEX and TNF-alpha. CONCLUSION: Our results suggest that not only angiogenin, TGF-beta, VEGF, APEX and TNF-alpha are significantly associated with the tumorigenesis, but also the close relationship between these factors might enhance the tumorigenesis of OSCC. We can not find clinical availability for diagnosis.
Subject(s)
Humans , Carcinogenesis , Carcinoma, Squamous Cell , Diagnosis , Electrophoresis, Agar Gel , Ethidium , Linear Models , Mouth Mucosa , Necrosis , RNA, Messenger , Sensitivity and Specificity , Transforming Growth Factor beta , Tumor Necrosis Factor-alpha , Vascular Endothelial Growth Factor AABSTRACT
Objective To investigate the expression of DNA damage and repair gene Apurinic/apyrimidinic endonuclease (APE1) protein in nasal NK/T cell lymphoma, and elucidate its clinical implication. Methods Expression of APE1 proteins was detected immunohistochemically in 10 normal lymph nodes and human nasal NK/T cells from lymphoma of 64 patients and their integral optical density was determined by means of image analytic system. The proliferation index and apoptosis index were determined by means of immunohistochemical staining and terminal dUTP nickend labeling (TUNEL) technique. Results 1. Nuclear, nucleus/cytoplasmic and cytoplasmic types of APE1 positive staining could be noted in nasal NK/T cell lymphoma. Expression of APE1 gene in nucleus was significantly strengthened compared to that in nucleus/cytoplasm and cytoplasm. In relapse or refractory group, no relapse or refractory group, and normal control group, the positive degree of cytoplasmic staining diminished significantly in the above order (P
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Objective To explore the relation between changes in apurinic/apyrimidinic endonuclease (APE1) gene expression and effects of melphalan on multiple myeloma (MM) cells. Methods Expression of APE1 protein was detected in MM cell line KM3 using immunocytochemical staining and Western blot assay after 0~15?mol/L melphalan treatment for 1~2d. Integral optical density was determined by means of image analysis system. Results There was positive relationship between levels of APE1 protein in KM3 cells and the treatment time and dose of melphalan. Conclusion Expression of APE1 protein could be induced by melphalan treatment. The result suggests that a high expression of APE1 protein may play a certain role in the resistance of multiple myeloma to melphalan.
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Objective To study the inhibitory effect of APE1 expression vector pSilence APE1 on growth of osteosarcoma in nude mice, and to elucidate the role of APE1 in pathogenesis and development of osteosarcoma. Methods Nude mice model bearing osteosarcoma was reproduced by implanting human osteosarcoma cell line 9901. Twenty mice were randomly divided into two groups: control group in which the mice were treated with lipofectamine, and experimental group in which the mice were treated with pSilence APE1. The tumor inhibition rate was calculated after 12 days of treatment. Meanwhile, the expression of APE1 protein, intratumor microvessel density (MVD), and proliferation index were observed by immunohistochemistry. Apoptosis index was assessed by terminal dUTP nick end labeling (TUNEL) technique. Results Down-regulation of APE1 expression of tumor cells was found in the group treated with pSilence APE1. The tumor inhibition rate was 38.23%. The intratumoral microvessel density (MVD) in experimental groups and proliferation index were significantly lower than the control group, while the apoptosis index was much higher. Conclusion Targeted knock down of APE1 by pSilence APE1 may inhibit the growth of osteosarcoma in vivo.
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Objective To construct DNA damage and repair gene apurinic/apyrimidinic endonuclease (APE1) siRNA expression vector pSilence APE1 and investigate its inhibitory effect on the expression of APE1 in osteosarcoma cell HOS and 9901 in vitro. Methods An expression plasmid of a short hairpin RNA target APE1, pSilence APE1 was constructed and transfected to 9901 and HOS cell by lipofectamine. The expression of APE1 protein in HOS and 9901 osteosarcoma cells posttransfected with pSilence APE1 was detected by immunohistochemistry. Meanwhile, the dose-effect and time-effect relationship of APE1 gene silence induced by pSilence APE1 were measured using Western blot analysis. Results After evaluation and sequencing, the APE1 siRNA expression vector pSilence APE1 was constructed successfully. The result of Western blotting and immunohistochemistry showed that APE1 protein in osteosarcoma cells could be knocked down specifically by pSilence APE1, and the inhibition rate of APE1 expression was 72%-95%. The best inhibition of expression of APE1 gene was 3.0 ?g and at the 72 h using pSilence APE1. Conclusion APE1 siRNA expression vector pSilence APE1 is successfully constructed that can significantly knock down APE1 gene expression in osteosarcoma cells.
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Objective To investigate the role of APE1 in the carcinogenesis and progression of colorectal carcinoma (CRC). Methods Expression of APE1 was determined with SP immunohistochemical technique in 40 specimens of normal colorectal mucosa, 60 specimens of colorectal mucosa adjacent to CRC, 72 specimens of colorectal adenoma, and 125 specimens of colorectal carcinoma. Results In normal colorectal mucosa, APE1 was detected in nuclei of epithelial cells. Shift of APE1 from nucleus to cytoplasm was observed in 6 of 60 (10%) specimens of mucosa adjacent to cancer. Such shift was observed in 92 of 125 (73.6%) CRC tissues and 60 of 72 (83.3%) colorectal adenoma, the incidence of both of them was significantly higher than that observed in normal colorectal mucosa and colorectal mucosa adjacent to CRC (P
ABSTRACT
Apurinic/apyrimidinic endonuclease/redox factor 1(APE/Ref 1) is a multifunctional protein that is responsible for repairing DNA damage,it also infleunce reduction oxidation (redox) reaction and modulate the DNA binding activity of transcription factors,it is important to cell survival.Recently, much data concerning APE1/Ref 1 and information on APE1/Ref 1 in a wide variety of functions and systems have rapid growth,including its gene,function,distribution and relations with some diseases,especially nervous system diseases and tumors.