Browsing by Author "Chan, Err-Cheng"

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An Informatics-assisted Label-free Approach for Personalized Tissue Membrane Proteomics: Case Study on Colorectal Cancer
(2011-04-01)
Han, Chia-Li
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Chen, Jinn-Shiun
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Chan, Err-Cheng
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Wu, Chien-Peng
;
Yu, Kun-Hsing
We developed a multiplexed label-free quantification strategy, which integrates an efficient gel-assisted digestion protocol, high-performance liquid chromatography tandem MS analysis, and a bioinformatics alignment method to determine personalized proteomic profiles for membrane proteins in human tissues. This strategy provided accurate (6% error) and reproducible (34% relative S. D.) quantification of three independently purified membrane fractions from the same human colorectal cancer (CRC) tissue. Using CRC as a model, we constructed the personalized membrane protein atlas of paired tumor and adjacent normal tissues from 28 patients with different stages of CRC. Without fractionation, this strategy confidently quantified 856 proteins (>= 2 unique peptides) across different patients, including the first and robust detection (Mascot score: 22,074) of the well-documented CRC marker, carcinoembryonic antigen 5 by a discovery-type proteomics approach. Further validation of a panel of proteins, annexin A4, neutrophils defensin A1, and claudin 3, confirmed differential expression levels and high occurrences (48-70%) in 60 CRC patients. The most significant discovery is the overexpression of stomatin-like 2 (STOML2) for early diagnostic and prognostic potential. Increased expression of STOML2 was associated with decreased CRC-related survival; the mean survival period was 34.77 +/- 2.03 months in patients with high STOML2 expression, whereas 53.67 +/- 3.46 months was obtained for patients with low STOML2 expression. Further analysis by ELISA verified that plasma concentrations of STOML2 in early-stage CRC patients were elevated as compared with those of healthy individuals (p < 0.001), suggesting that STOML2 may be a noninvasive serological biomarker for early CRC diagnosis. The overall sensitivity of STOML2 for CRC detection was 71%, which increased to 87% when combined with CEA measurements. This study demonstrated a sensitive, label-free strategy for differential analysis of tissue membrane proteome, which may provide a roadmap for the subsequent identification of molecular target candidates of multiple cancer types. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.003087, 1-15, 2011.
Detection of autoantibodies against Rabphilin-3A-like protein as a potential biomarker in patient's sera of colorectal cancer
(2011-07-15)
Chen, Jinn-Shiun
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Kuo, Yung-Bin
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Chou, Yeh-Pin
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Chan, Chung-Chuan
;
Fan, Chung-Wei
Background: Rabphilin-3A-like (RPH3AL) protein functions in the regulation of hormone exocytosis, and mutations in the RPHA3L gene have been associated with tumorigenesis in colorectal cancer (CRC). We evaluated the potential use of anti-RPH3AL autoantibodies as a marker for CRC detection. Methods: Sera from 84 patients with CRC and 63 healthy controls were analysed for the presence of RPH3AL autoantibodies with a Western blotting assay. Results: The frequencies of RPH3AL autoantibodies in the early stage, advanced stage and all CRC patients were 64.7%, 78.0% and 72.6%, respectively. These values are significantly higher than the frequency of RPH3AL autoantibodies in healthy controls (15.9%, P<0.001). Although the presence of RPH3AL autoantibodies did not correlate with clinical parameters, RPH3AL autoantibodies were found in 69.4% (34/49) of CRC patients who were negative for carcinoembryonic antigen. The value of the area under the receiver operating characteristic curve of RPH3AL autoantibody was 0.84, which suggests that screening for these autoantibodies could potentially be used for CRC diagnosis. Conclusion: Circulating RPH3AL autoantibodies are prevalent in patients with CRC, and detection of these autoantibodies might provide a novel non-invasive approach for CRC diagnosis. (C) 2011 Elsevier B.V. All rights reserved.
Identification and clinical association of anti-cytokeratin 18 autoantibody in COPD
(2010-02-16)
Kuo, Yung-Bin
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Chang, C. Allen
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Wu, Yao-Kuang
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Hsieh, Meng-Jer
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Tsai, Chung-Hsien
The etiology of chronic obstructive pulmonary disease (COPD) remains unclear. A mechanism involving the autoimmune reaction in the pathogenesis of COPD has been proposed but not confirmed. The aim of this study was to investigate whether serum autoantibodies against pulmonary cellular proteins are present in COPD patients and to identify their autoantigens if possible. Samples from 50 COPD patients and 42 control subjects were studied. Circulating autoantibodies were detected by Western blot. Immunoprecipitation and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry were used to identify the autoantigens. Autoantibodies against pulmonary cellular antigens were found in the sera of COPD patients. Specifically, an autoantibody against the 45-kDa human cytokeratin 18 protein was found in 76.0% of COPD patients and 23.8% of control subjects (p < 0.001). Furthermore, the cytokeratin 18 autoantibody level was positively correlated with the FEV(1) (L) (p = 0.013) and FEV(1) (%pred.) (p, = 0.043) values observed in COPD patients. This study identified the pulmonary epithelial cytokeratin 18 protein as a COPD-associated autoantigen and found that anti-cytokeratin 18 autoantibodies were prevalent in COPD patients. Our results support the hypothesis that humoral autoimmunity may be involved in the pathogenesis of COPD. (C) 2009 Elsevier B.V. All rights reserved.
Identification of Phospholipid Scramblase 1 as a Biomarker and Determination of Its Prognostic Value for Colorectal Cancer
(2011-01-01)
Kuo, Yung-Bin
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Chan, Chung-Chuan
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Chang, C. Allen
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Fan, Chung-Wei
;
Hung, Ray-Ping
The purpose of this study was to examine the expression of phospholipid scramblase 1 (PLSCR1) in tumor tissues and plasma specimens of patients with colorectal cancer (CRC), as well as analyze its association with clinical parameters. The expression levels of PLSCR1 protein in 104 matched CRC and adjacent normal tissue sections and 50 pairs of CRC tissue blocks were determined by use of immunohistochemical and Western blot analyses, respectively. To evaluate the diagnostic potential of PLSCR1, the plasma levels of PLSCR1 were investigated in 111 additional subjects (59 CRC patients and 52 healthy controls) by Western blot. PLSCR1 was overexpressed in malignant adenocarcinoma tissues compared with normal colorectal mucosa (P < 0.001). In addition, the plasma level of PLSCR1 was not only significantly elevated in CRC patients compared with healthy individuals (P < 0.001), but it was also substantially increased in early stage CRC (P < 0.001). Importantly, the overall sensitivity and specificity of PLSCR1 for CRC detection were 80% and 59.6%, respectively. The area under the ROC curve of PLSCR1 for CRC diagnosis is 0.75, which increases to 0.8 if combined with the measurement of carcinoembryonic antigen. Univariate analysis with the Cox regression model revealed that elevated PLSCR1 expression indicated a poor prognosis for CRC, This study showed that PLSCR1 protein levels were significantly elevated in both the cancer tissue and plasma of CRC patients. Moreover, the plasma levels of PLSCR1 were significantly elevated in patients with early stage CRC compared with healthy individuals, suggesting that PLSCR1 might be used as a noninvasive serological diagnostic and prognostic biomarker for CRC. (c) 2011 The Feinstein Institute for Medical Research, www.feinsteininstitute.org Online address: hffp://www.molmed.org doi: 10.2119/molmed.2010.00115
Identification of SEC61 beta and its autoantibody as biomarkers for colorectal cancer
(2011-05-12)
Fan, Chung-Wei
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Chan, Chung-Chuan
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Chen, Kuei-Tien
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Twu, Jen
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Huang, Ya-Shu
Background: To identify novel serological biomarkers for human colorectal cancer (CRC). we analyzed CRC tissues using gel-assisted digestion and isobaric tags with related and absolute quantitation (iTRAQ) labeling mass spectrometry (MS). By comparing pairs of tumor tissues and matched normal tissues. we discovered the SEC61 beta with expression changes 3.3-fold and a marginal statistical significance (p=0.052) previously. Methods: SEC61 beta expression in CRC tissues was further analyzed by western blotting and immunohistochemistry. We next assessed the putative diagnostic value of the SEC61 beta autoantibody as a serum marker. Results: Using western blotting analysis. SEC61 beta expression was increased 1.9-fold in tumor tissues. Immunohistochemical analysis of 64 CRC specimens showed that SEC61 beta was positively detected in 64% of the tumors, but weakly or not detected in > 80% of the adjacent nontumor epithelial cells. Western blot analysis with plasma samples showed that the sensitivity and specificity of the SEC61 beta autoantibody from patients with CRC were 79% and 75%. respectively. Importantly, the results of the SEC61 beta autoantibody for early detection of colorectal cancer revealed a higher sensitivity of 77% than the carcinoembryonic antigen (CEA) assay. Conclusions: Measurement of SEC61 beta autoantibody levels may provide an alternative detection indicator for CRC, particularly among early-stage patients. (C) 2011 Elsevier B.V. All rights reserved.
Multiple serological biomarkers for colorectal cancer detection
(2010-04-01)
Chan, Chung-Chuan
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Fan, Chung-Wei
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Kuo, Yung-Bin
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Chen, Yu-Han
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Chang, Pi-Yueh
The aim of this study was to initiate a survey of human autoantibody responses to a panel of select colorectal tumor-associated antigens identified by previous serological analysis of a cDNA expression library and to subsequently identify multiple serological biomarkers for the detection of colorectal cancer. For screening of autoantibodies against colorectal tumor-associated antigens, sera from 94 colorectal cancer patients and 54 normal controls were analyzed by enzyme-linked immunosorbent assay using recombinant rCCCAP, rHDAC5, rP53, rNMDAR and rNY-CO-16 proteins as coating antigens. Seropositivity among colorectal cancer patients to the 5 individual coating antigens varied from 18.1% to 35.1%. Seropositivity to any of the 5 coating antigens was 58.5% and combining this analysis with evaluation of serum carcinoembryonic antigen (>= 5 ng/ml) significantly increased the seropositivity to 77.6%. Seropositivity of early-stage (Dukes' Stages A and B) colorectal cancer patients to CEA was 21.9%, and seropositivity to any of the 5 colorectal cancer-associated antigens was 53.7%, and the combination of these 2 measurements resulted in a higher diagnostic capacity (65.9%) than either marker alone. In conclusion, these results collectively indicated that combined detection of serum autoantibody profiles against our panel of colorectal tumor-associated antigens and the analysis of carcinoembryonic antigen provides a promising diagnostic biomarker for colorectal cancer, particularly among early-stage patients.
Performance of rapid-test kits for the detection of the pandemic influenza A/H1N1 virus
(2011-05-01)
Tsao, Kuo-Chien
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Kuo, Yung-Bin
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Huang, Chung-Guei
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Chau, Shao-Wen
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Chan, Err-Cheng
The early detection of pandemic influenza strains is a key factor for clinicians in treatment decisions and infection control practices. The aims of this study were to determine the analytical sensitivity and clinical performance of the commercially available influenza rapid tests in Taiwan. Four rapid tests for influenza virus (BinaxNow test, QuickVue test, TRU test, and Formosa Rapid test) were evaluated for their detection limit against four influenza viruses (the 2009 pandemic influenza A virus H1N1, seasonal influenza virus H1N1, H3N2, and influenza B virus) circulating in Taiwan. The viral load of these isolates were quantified by rtRT-PCR and then diluted 2-fold serially for the comparison. The lowest detectable viral load of the pandemic influenza A virus H1N1 by the Formosa Rapid test, QuickVue test, TRU test, and Binax Now test was 5.3 x 10(4), 1.0 x 10(5), 1.0 x 10(5), and 4.2 x 10(5) copies/mu l, respectively. Of these four tests, the two most sensitive tests (the QuickVue test and the Formosa Rapid test) were chosen to evaluate 62 nasopharyngeal specimens from patients who were suspected of infection with pandemic influenza A virus H1N1. The positive rate for the Formosa Rapid test and the QuickVue test were 53.2% (33/62) and 45.2% (28/62) (McNemar's test, P = 0.125), respectively. In conclusion, the Formosa Rapid test was the most sensitive test in the present study for the detection of influenza antigens and its clinical performance was similar to that of the QuickVue test (Kappa = 0.776). This suggests that the Formosa Rapid test could be used to aid clinical decision making in primary health care settings during outbreaks of influenza. (C) 2011 Elsevier B.V. All rights reserved.
Prevalence of Human Papillomavirus Genotypes in Northern Taiwanese Women
(2010-10-01)
Tsao, Kuo-Chien
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Huang, Chung-Guei
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Kuo, Yung-Bin
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Chang, Ting-Chang
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Sun, Chien-Feng
The prevalence of Human Papillomavirus (HPV) in the general population of northern Taiwan is described. A total of 343 consecutive cervical swabs from women visiting the medical center for routine gynecologic care were included. Cervical cell cytology was examined by the Papanicolaou (Pap) test, and a PCR-based hybridization gene chip analysis was used to identify HPV genotypes. The HPV prevalence in the overall population was 32.4%. When divided into two groups according to cytology, 20.9% of women with normal cytology were HPV positive while 75.3% of women with abnormal cytology were HPV positive. Among positive samples, 68.5% were single type infections while 31.5% harbored multiple HPV types. A total of 32 types of HPV were identified; the leading five were HPV16 (5.8%), HPV58 (5.3%), HPV53 (4.1%), HPV52 (3.8%), and HPV18 (2.3%). Our results constitute baseline data and may provide important implications for future prophylactic programs. The relatively high prevalence of HPV 58, 53, and 52 among northern Taiwanese women has important implications for vaccine development. J. Med. Virol. 82:1739-1745, 2010. (C) 2010 Wiley-Liss, Inc.
Quantitative detection of survivin in malignant pleural effusion for the diagnosis and prognosis of lung cancer
(2009-01-18)
Wu, Yao-Kuang
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Chen, Kuei-Tien
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Kuo, Yung-Bin
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Huang, Ya-Shu
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Chan, Err-Cheng
in the present study, pleural effusions are the first time to be used as the specimens for detection of survivin expression in lung cancer patients. We demonstrated that by quantifying survivin expression with enzyme-linked immunosorbent assay (ELISA) in the 80 effusion samples exhibited a diagnostic power of 85% and 75% in sensitivity and specificity, respectively. A multivariate analysis with the Cox regression model revealed that both high survivin expression and cancer cells of stage IV were the indicators for poor prognosis of lung cancer. In conclusion, quantitative assay of survivin in pleural effusion could be useful both in diagnosis and prognosis for lung cancer. (C) 2008 Elsevier Ireland Ltd. All rights reserved.
Quantitative detection of survivin in malignant pleural effusion for the diagnosis and prognosis of lung cancer
(2009-01-18)
Wu, Yao-Kuang
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Chen, Kuei-Tien
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Kuo, Yung-Bin
;
Huang, Ya-Shu
;
Chan, Err-Cheng
in the present study, pleural effusions are the first time to be used as the specimens for detection of survivin expression in lung cancer patients. We demonstrated that by quantifying survivin expression with enzyme-linked immunosorbent assay (ELISA) in the 80 effusion samples exhibited a diagnostic power of 85% and 75% in sensitivity and specificity, respectively. A multivariate analysis with the Cox regression model revealed that both high survivin expression and cancer cells of stage IV were the indicators for poor prognosis of lung cancer. In conclusion, quantitative assay of survivin in pleural effusion could be useful both in diagnosis and prognosis for lung cancer. (C) 2008 Elsevier Ireland Ltd. All rights reserved.