Proteomic analysis of cholangiocarcinoma cell line

Srisomsap, Chantragan; Sawangareetrakul, Phannee; Subhasitanont, Pantipa; Panichakul, Tasanee; Keeratichamroen, Siriporn; Lirdprapamongkol, Kriengsak; Chokchaichamnankit, Daranee; Sirisinha, Stitaya; Svasti, Jisnuson

doi:10.1002/pmic.200300651

Cited by 62 publications

(56 citation statements)

References 25 publications

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“…Cytokeratin is up-regulated in breast cancer (19), especially in neu-overexpressing breast cancer (20). Cytokeratin was also overexpressed in hepatocarcinoma, cervical cancer, and renal cancer, similar tissues as we show here in the murine system (21)(22)(23). Gsn, as a tumor suppressor, is down-regulated significantly in human breast and ovarian cancers (9,11,19,24).…”

Section: Resultssupporting

confidence: 60%

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The Tumor Antigen Repertoire Identified in Tumor-Bearing Neu Transgenic Mice Predicts Human Tumor Antigens

Knutson

Gad

et al. 2006

Cancer Research

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FVB/N mice transgenic for nontransforming rat neu develop spontaneous breast cancers that are neu positive and estrogen receptor negative, mimicking premenopausal human breast cancer. These animals have been widely used as a model for immunobased therapies targeting HER-2/neu. In this study, we used serological analysis of recombinant cDNA expression libraries to characterize the antigenic repertoire of neu transgenic (neu-tg) mice and questioned the ability of this murine model to predict potential human tumor antigens. After screening 3 Â 10 6 clones from 3 different cDNA libraries, 15 tumor antigens were identified, including cytokeratin 2-8, glutamyl-prolyl-tRNA synthetase, complement C3, galectin 8, and serine/threonine-rich protein kinase 1. Multiple proteins involved in the Rho/Rho-associated, coiled coil-containing protein kinase (Rock) signal transduction pathway were found to be immunogenic, including Rock1, Rho/Rac guanine nucleotide exchange factor 2, and schistosoma mansoni adult worm antigen preparation 70. All of the identified antigens are self-proteins that are expressed in normal tissues in addition to breast tumors and the majority of the antigens are intracellular proteins. More than half of the mouse tumor antigens have human homologues that have been reported previously as tumor antigens. Finally, the tumor-specific antibody immunity and marked immune cell infiltration that was observed in mice with spontaneous tumors were not observed in mice with transplanted tumors. Our results indicate that neu-tg mice bearing spontaneous tumors develop humoral immunity to their tumors similar to cancer patients and that tumor antigens identified in transgenic mouse may predict immunogenic human homologues.

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Section: Resultssupporting

confidence: 60%

“…3). It is also reported to be overexpressed in cancer patients (19)(20)(21)(22)(23). Rock1 is the downstream effector of Rho and regulates actin polymerization and focal adhesion (Fig.…”

Section: Discussionmentioning

confidence: 99%

The Tumor Antigen Repertoire Identified in Tumor-Bearing Neu Transgenic Mice Predicts Human Tumor Antigens

Knutson

Gad

et al. 2006

Cancer Research

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“…The proteomic and secretomic maps of HuCCA-1 cell line, a Thai human cholangiocarcinoma cell line were described by our laboratory (11). The cell line has been used for screening the cytotoxic activities of Thai medicinal plants and proteomic profiling upon treatment with pomiferin, a prenylated isoflavone from Derris malaccensis (24).…”

Section: Discussionmentioning

confidence: 99%

“…The risk of cholangiocarcinoma increases in patients with chronic liver disease with either form of viral hepatitis, B or C (7,8), alcoholic liver disease or cirrhosis from a number of causes (9,10). Our group has established the proteomic map of a Thai human cholangiocarcinoma HuCCA-1 cell line and compared it to Thai human hepatocellular carcinoma HCC-S102 cell line and hepatoblastoma HepG2 cell line by studying their soluble proteins (11) and membrane proteins (12).…”

Section: Introductionmentioning

confidence: 99%

Apigenin inhibits growth and induces apoptosis in human cholangiocarcinoma cells

et al. 2017

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Abstract.A promising nutraceutical, apigenin, was recently revealed to exhibit biological activity in inhibiting several types of cancer. The effects of apigenin on the growth inhibition and apoptosis of the cholangiocarcinoma HuCCA-1 cell line were investigated. Protein alterations subsequent to apigenin treatment were studied using a proteomic approach. The values of 20, 50 and 90% inhibition of cell growth (IC 20 , IC 50 and IC 90 ) were determined by MTT cell viability assay. Apoptotic cell death was detected using two different methods, a flow cytometric analysis (Muse Cell Analyzer) and DNA fragmentation assay. A number of conditions including attached and detached cells were selected to perform two-dimensional gel electrophoresis (2-DE) to study the alterations in the expression levels of treated and untreated proteins and identified by liquid chromatography (LC)/tandem mass spectrometry (MS/MS). The IC 20 , IC 50 and IC 90 values of apigenin after 48 h treatment in HuCCA-1 cells were 25, 75 and 200 µM, respectively, indicating the cytotoxicity of this compound. Apigenin induced cell death in HuCCA-1 cells via apoptosis as detected by flow cytometric analysis and exhibited, as confirmed with DNA fragmentation, characteristics of apoptotic cells. A total of 67 proteins with altered expression were identified from the 2-DE analysis and LC/MS/MS. The cleavage of proteins involved in cytoskeletal, cytokeratin 8, 18 and 19, and high expression of S100-A6 and S100-A11 suggested that apoptosis was induced by apigenin via the caspase-dependent pathway. Notably, two proteins, heterogeneous nuclear ribonucleoprotein H and A2/B1, disappeared completely subsequent to treatment, suggesting the role of apigenin in inducing cell death. The present study indicated that apigenin demonstrates an induction of growth inhibition and apoptosis in cholangiocarcinoma cells and the apoptosis pathway was confirmed by proteomic analysis.

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“…In a more recent study using a 2D-DIGE-MS approach, the protein expression profiles of primary cultured hepatocytes and of HCC cell lines were compared (Fujii et al, 2005); in this study, 44 out of 1238 protein spots detected were differentially expressed in HCC cell lines. Proteomic expression studies using a variety of approaches such as 2D-PAGE-MS, 2D-DIGE-MS, AACT-1D PAGE-MALDI-TOF/TOF-MS/MS and 1D PAGE-ICAT-LC-ESI-MS/MS have also been performed to compare protein profiles of alpha-fetoprotein (AFP)-producing and non-AFP-producing HCC cell lines (Yokoo et al, 2004), protein profiles of HCC cell lines with different metastatic potentials Ding et al, 2004;Yu et al, 2004;Shui et al, 2005) or protein profiles of cholangiocarcinoma and HCC cell lines (Srisomsap et al, 2004). As a result, 11 proteins involved in apoptosis, glucose metabolism and cytoskeletal organization have been identified as differentially expressed between AFP-producing and non-AFP-producing cell lines.…”

Section: Cell Linesmentioning

confidence: 99%