Journal: Journal of Thoracic Oncology

Article Title: AZGP1 Autoantibody Predicts Survival and Histone Deacetylase Inhibitors Increase Expression in Lung Adenocarcinoma

doi: 10.1097/jto.0b013e318189f5ec

Figure Lengend Snippet: FIGURE 1. Levels of AZGP1 auto- antibody detected in (A) AD and (B) control subjects. The first 140 AD patients showed an autoantibody level of 2 whereas the first 139 control patients showed a level 2. A similar number of patients in each subset showed a level of AZGP1 au- toantibody that was 2. AD, lung adenocarcinoma.

Article Snippet: Immunohistochemical staining was performed on the DAKO Autostainer (DAKO, Carpinteria, CA) using DAKO LSAB and diaminobenzadine as the chromogen, as previously described.9 Deparaffinized sections of the TMA prepared as previously described,10 at 5- m thickness were labeled with AZGP1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, Cat SC-13585, mouse, 1:400) after microwave antigen retrieval in 10 mM sodium citrate buffer, pH 6.

Techniques: Control

Journal: Journal of Thoracic Oncology

Article Title: AZGP1 Autoantibody Predicts Survival and Histone Deacetylase Inhibitors Increase Expression in Lung Adenocarcinoma

doi: 10.1097/jto.0b013e318189f5ec

Figure Lengend Snippet: FIGURE 2. A, Kaplan-Meier survival analysis for AZGP1 au- toantibody in AD patients. Patients with high levels of AZGP1 autoantibody (level median 1.02) had significantly better 5-year survival than patients with low levels of AZGP1 autoantibody (p 0.002). B, Kaplan-Meier survival curve analysis for AZGP1 autoantibody production in patients with stage I and II AD. Patients with high levels (level median 1.02) of autoantibody had significantly better 5-year survival than patients with low levels (p 0.008). AD, lung adeno- carcinoma.

Article Snippet: Immunohistochemical staining was performed on the DAKO Autostainer (DAKO, Carpinteria, CA) using DAKO LSAB and diaminobenzadine as the chromogen, as previously described.9 Deparaffinized sections of the TMA prepared as previously described,10 at 5- m thickness were labeled with AZGP1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, Cat SC-13585, mouse, 1:400) after microwave antigen retrieval in 10 mM sodium citrate buffer, pH 6.

Techniques:

Journal: Journal of Thoracic Oncology

Article Title: AZGP1 Autoantibody Predicts Survival and Histone Deacetylase Inhibitors Increase Expression in Lung Adenocarcinoma

doi: 10.1097/jto.0b013e318189f5ec

Figure Lengend Snippet: FIGURE 3. A, Verification of Kaplan-Meier survival analysis for AZGP1 autoantibody in AD patients. Patients with high levels of AZGP1 autoantibody (level median 1.02) has sig- nificantly better 5-year survival than patients with low levels of autoantibody (p 0.02). B, Verification of Kaplan-Meier survival analysis for AZGP1 autoantibody production in patients with stage I and II AD. Patients with high levels (level median 1.02) of autoantibody had significantly bet- ter 5-year survival than patients with low levels (p 0.036). AD, lung adenocarcinoma.

Article Snippet: Immunohistochemical staining was performed on the DAKO Autostainer (DAKO, Carpinteria, CA) using DAKO LSAB and diaminobenzadine as the chromogen, as previously described.9 Deparaffinized sections of the TMA prepared as previously described,10 at 5- m thickness were labeled with AZGP1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, Cat SC-13585, mouse, 1:400) after microwave antigen retrieval in 10 mM sodium citrate buffer, pH 6.

Techniques:

Journal: Journal of Thoracic Oncology

Article Title: AZGP1 Autoantibody Predicts Survival and Histone Deacetylase Inhibitors Increase Expression in Lung Adenocarcinoma

doi: 10.1097/jto.0b013e318189f5ec

Figure Lengend Snippet: FIGURE 4. Oligonucleotide mi- croarray analysis of 86 AD and 10 normal samples revealed that 31.3% of adenocarcinomas ex- pressed AZGP1 at least two fold more than the controls. Samples 1 to 86, AD; samples 87 to 96, con- trols. AD, lung adenocarcinoma.

Article Snippet: Immunohistochemical staining was performed on the DAKO Autostainer (DAKO, Carpinteria, CA) using DAKO LSAB and diaminobenzadine as the chromogen, as previously described.9 Deparaffinized sections of the TMA prepared as previously described,10 at 5- m thickness were labeled with AZGP1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, Cat SC-13585, mouse, 1:400) after microwave antigen retrieval in 10 mM sodium citrate buffer, pH 6.

Techniques:

Journal: Journal of Thoracic Oncology

Article Title: AZGP1 Autoantibody Predicts Survival and Histone Deacetylase Inhibitors Increase Expression in Lung Adenocarcinoma

doi: 10.1097/jto.0b013e318189f5ec

Figure Lengend Snippet: FIGURE 5. AZGP1 protein staining was not detected in (A) normal lung tissues, but was seen in (B) and (C) 42.8% of AD, and (D) 8.3% of SCC by immunohistochemistry. AD, lung adenocarcinoma; SCC, lung squamous cell carcinoma.

Article Snippet: Immunohistochemical staining was performed on the DAKO Autostainer (DAKO, Carpinteria, CA) using DAKO LSAB and diaminobenzadine as the chromogen, as previously described.9 Deparaffinized sections of the TMA prepared as previously described,10 at 5- m thickness were labeled with AZGP1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, Cat SC-13585, mouse, 1:400) after microwave antigen retrieval in 10 mM sodium citrate buffer, pH 6.

Techniques: Staining, Immunohistochemistry

Journal: Journal of Thoracic Oncology

Article Title: AZGP1 Autoantibody Predicts Survival and Histone Deacetylase Inhibitors Increase Expression in Lung Adenocarcinoma

doi: 10.1097/jto.0b013e318189f5ec

Figure Lengend Snippet: FIGURE 6. A, Western blot demonstrating AZGP1 protein in the serum of all patients, irrespective of AZGP1 protein staining on tissue microarrays. AD, patient with lung adenocarcinoma; N, normal control subject; NE, no AZGP1 protein expression on TMA (negative); WE, weak AZGP1 expression (1); IE, intermediate AZGP1 expression (2); SE, strong AZGP1 expression (3). B, Western blot demonstrating AZGP1 protein in the serum of all patients, irrespective of AZGP1 autoantibody produc- tion. AD, patient with lung adenocarcinoma; N, normal control subject; BL, patient with benign lung disease; Low, patient with serum autoantibody level 2; High, patient with serum autoantibody level 2.

Article Snippet: Immunohistochemical staining was performed on the DAKO Autostainer (DAKO, Carpinteria, CA) using DAKO LSAB and diaminobenzadine as the chromogen, as previously described.9 Deparaffinized sections of the TMA prepared as previously described,10 at 5- m thickness were labeled with AZGP1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, Cat SC-13585, mouse, 1:400) after microwave antigen retrieval in 10 mM sodium citrate buffer, pH 6.

Techniques: Western Blot, Staining, Control, Expressing

Journal: Journal of Thoracic Oncology

Article Title: AZGP1 Autoantibody Predicts Survival and Histone Deacetylase Inhibitors Increase Expression in Lung Adenocarcinoma

doi: 10.1097/jto.0b013e318189f5ec

Figure Lengend Snippet: FIGURE 7. A, AZGP1 mRNA pro- duction is up-regulated in a time dependent manner by HDAC inhibi- tor TSA in A549 cells. Gene pro- moter demethylation by 5-AZA has no effect on AZGP1 mRNA expres- sion in A549 and attenuates the up- regulation by TSA. B, AZGP1 mRNA production is up-regulated in a time dependent manner by HDAC inhibi- tor TSA in SKLU1 cells. Gene pro- moter demethylation by 5-AZA has no effect on AZGP1 mRNA expres- sion in SKLU1 cells and attenuates the up-regulation by TSA. TSA, tri- chostatin A; 5-AZA, 5-aza-2’ deoxy- cytidine. *p 0.05 with t test as compared with control.

Article Snippet: Immunohistochemical staining was performed on the DAKO Autostainer (DAKO, Carpinteria, CA) using DAKO LSAB and diaminobenzadine as the chromogen, as previously described.9 Deparaffinized sections of the TMA prepared as previously described,10 at 5- m thickness were labeled with AZGP1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, Cat SC-13585, mouse, 1:400) after microwave antigen retrieval in 10 mM sodium citrate buffer, pH 6.

Techniques: Control

Journal: OncoTargets and therapy

Article Title: Reduction of AZGP1 predicts poor prognosis in esophageal squamous cell carcinoma patients in Northern China

doi: 10.2147/OTT.S113932

Figure Lengend Snippet: Primer sequences used for quantitative polymerase chain reaction analyses

Article Snippet: Blocked by 10% normal mouse serum at room temperature for 30 minutes, the slides were then incubated with primary anti-AZGP1 monoclonal antibody (Proteintech; 1:200 dilution) at 4°C overnight, which has been confirmed by Western blot assay ( ).

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: OncoTargets and therapy

Article Title: Reduction of AZGP1 predicts poor prognosis in esophageal squamous cell carcinoma patients in Northern China

doi: 10.2147/OTT.S113932

Figure Lengend Snippet: Downregulation of AZGP1 in ESCC tumor tissues. Notes: ( A , B ) Quantitative real-time polymerase chain reaction was used to compare AZGP1 expression between tumor and nontumor tissues in 45 ESCCs. The result was normalized by β-actin. Each sample was analyzed in triplicate. *** P <0.01. Abbreviation: ESCC, esophageal squamous cell carcinoma.

Article Snippet: Blocked by 10% normal mouse serum at room temperature for 30 minutes, the slides were then incubated with primary anti-AZGP1 monoclonal antibody (Proteintech; 1:200 dilution) at 4°C overnight, which has been confirmed by Western blot assay ( ).

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: OncoTargets and therapy

Article Title: Reduction of AZGP1 predicts poor prognosis in esophageal squamous cell carcinoma patients in Northern China

doi: 10.2147/OTT.S113932

Figure Lengend Snippet: Representative of AZGP1 expression in two pairs of ESCC (below) and adjacent normal tissue (above) were detected by immunostaining with anti-AZGP1 antibody (brown). Notes: The slides were counterstained with hematoxylin. Original magnification: ×100 (left) and ×200 (right). Abbreviation: ESCC, esophageal squamous cell carcinoma.

Article Snippet: Blocked by 10% normal mouse serum at room temperature for 30 minutes, the slides were then incubated with primary anti-AZGP1 monoclonal antibody (Proteintech; 1:200 dilution) at 4°C overnight, which has been confirmed by Western blot assay ( ).

Techniques: Expressing, Immunostaining

Journal: OncoTargets and therapy

Article Title: Reduction of AZGP1 predicts poor prognosis in esophageal squamous cell carcinoma patients in Northern China

doi: 10.2147/OTT.S113932

Figure Lengend Snippet: Association of AZGP1 expression with clinicopathologic features of patients with esophageal squamous cell carcinoma (n=215)

Article Snippet: Blocked by 10% normal mouse serum at room temperature for 30 minutes, the slides were then incubated with primary anti-AZGP1 monoclonal antibody (Proteintech; 1:200 dilution) at 4°C overnight, which has been confirmed by Western blot assay ( ).

Techniques: Expressing, Cell Differentiation

Journal: OncoTargets and therapy

Article Title: Reduction of AZGP1 predicts poor prognosis in esophageal squamous cell carcinoma patients in Northern China

doi: 10.2147/OTT.S113932

Figure Lengend Snippet: Survival curves plotted by employing Kaplan–Meier estimator. Notes: ( A ) Kaplan–Meier plot with univariate analysis indicated that patients with low AZGP1 expression had a poor DSS ( P <0.01) than those with normal AZGP1 expression, log-rank test. ( B ) A stratified survival analysis according to the pathological stage revealed that AZGP1 expression discernibility on DSS was only pronounced in patients with advanced clinical stage (stage I–II; P <0.001), log-rank test. ( C , D ) Kaplan–Meier plot with univariate analysis indicated that patients with low AZGP1 expression had a poorer local recurrence-free ( P =0.016) and metastasis-free survival ( P =0.014) than those with normal AZGP1 expression, log-rank test. Abbreviation: DSS, disease-specific survival.

Article Snippet: Blocked by 10% normal mouse serum at room temperature for 30 minutes, the slides were then incubated with primary anti-AZGP1 monoclonal antibody (Proteintech; 1:200 dilution) at 4°C overnight, which has been confirmed by Western blot assay ( ).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: Reduction of AZGP1 predicts poor prognosis in esophageal squamous cell carcinoma patients in Northern China

doi: 10.2147/OTT.S113932

Figure Lengend Snippet: Univariate log-rank analysis for clinicopathologic features and AZGP1 expression

Article Snippet: Blocked by 10% normal mouse serum at room temperature for 30 minutes, the slides were then incubated with primary anti-AZGP1 monoclonal antibody (Proteintech; 1:200 dilution) at 4°C overnight, which has been confirmed by Western blot assay ( ).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: Reduction of AZGP1 predicts poor prognosis in esophageal squamous cell carcinoma patients in Northern China

doi: 10.2147/OTT.S113932

Figure Lengend Snippet: Multivariate analysis

Article Snippet: Blocked by 10% normal mouse serum at room temperature for 30 minutes, the slides were then incubated with primary anti-AZGP1 monoclonal antibody (Proteintech; 1:200 dilution) at 4°C overnight, which has been confirmed by Western blot assay ( ).

Techniques: Expressing