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Identification of GLI1 direct target genes in human pancreatic cancer cell line Panc-1

ABSTRACT: Panc-1GLI1ER and Panc-1ER were established from human pancreatic cancer cell line Panc-1. Panc-1GLI1ER cells express a chimeric transgene composed of an enhanced green fluorescent protein (EGFP)-tagged version of the amino-terminal half of GLI1, fused to the AF2 domain of the mouse estrogen receptor 1(Esr1) cDNA. On the other hand, Panc-1ER harbor only EGFP-tagged AF2 domain. To identify GLI1 direct target genes, total RNAs were purified from the cells before and 3 hours after the beta-estradiol (E2) treatment. Gene expression profiles were analyzed by AGILENT human 4x44 cDNA microarray. Keywords: Expression profiling by genome tiling array GLI1 direct target genes were identified in Panc-1 cells. Gene expression profiles of Panc-1GLI1ER and Panc-1ER cells were analyzed by AGILENT human 4x44 cDNA microarray before and 3 hours after treatment of beta-estradiol (E2) at doses of 10nM.

ORGANISM(S): Homo sapiens

SUBMITTER: Inaguma Shingo

PROVIDER: S-ECPF-GEOD-28901 | biostudies-other |

REPOSITORIES: biostudies-other

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Identification of GLI1 direct target genes in human pancreatic cancer cell line Panc-1

Project description:Panc-1GLI1ER and Panc-1ER were established from human pancreatic cancer cell line Panc-1. Panc-1GLI1ER cells express a chimeric transgene composed of an enhanced green fluorescent protein (EGFP)-tagged version of the amino-terminal half of GLI1, fused to the AF2 domain of the mouse estrogen receptor 1(Esr1) cDNA. On the other hand, Panc-1ER harbor only EGFP-tagged AF2 domain. To identify GLI1 direct target genes, total RNAs were purified from the cells before and 3 hours after the beta-estradiol (E2) treatment. Gene expression profiles were analyzed by AGILENT human 4x44 cDNA microarray. Keywords: Expression profiling by genome tiling array GLI1 direct target genes were identified in Panc-1 cells. Gene expression profiles of Panc-1GLI1ER and Panc-1ER cells were analyzed by AGILENT human 4x44 cDNA microarray before and 3 hours after treatment of beta-estradiol (E2) at doses of 10nM.

2013-07-18 | E-GEOD-28901 | biostudies-arrayexpress

Identification of GLI1 direct target genes in human pancreatic cancer cell line Panc-1

2013-07-18 | GSE28901 | GEO

Identification of GLI1 direct and indirect target genes in human pancreatic cancer cell line Panc-1

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Identification of GLI1 direct target genes in human prostate cancer cell line PC3

Project description:PC3GLI1ER and PC3ER were established from human prostate cancer cell line PC3. PC3GLI1ER cells express a chimeric transgene composed of an enhanced green fluorescent protein (EGFP)-tagged version of the amino-terminal half of GLI1, fused to the AF2 domain of the mouse estrogen receptor 1(Esr1) cDNA. On the other hand, PC3ER harbor only EGFP-tagged AF2 domain. To identify GLI1 direct target genes, total RNAs were purified from the cells before and 3 hours after the beta-estradiol (E2) treatment. Gene expression profiles were analyzed by AGILENT human 4x44 cDNA microarray. Keywords: Expression profiling by array

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Identification of ZIC2 target genes in human pancreatic cancer cell lines PANC-1 and KP-4

Project description:PANC-1Tet/ZIC2 and PANC-1Tet/empty were established from human pancreatic cancer cell line PANC-1. PANC-1Tet/ZIC2 cells express FLAG-tagged human ZIC2 on the withdrawal of DOX. On the other hand, PANC-1Tet/empty was transfected an empty vector for the control experiment. To identify ZIC2 target genes, total RNAs were purified from the cells before and 48 hours after the DOX withdrawal. Gene expression profiles were analyzed by AGILENT human 4x44k cDNA microarray. As well as ZIC2-inducible system, we performed ZIC2-knockdown experiments in PANC-1 human pancreatic cancer cells. After 96 hours transfection of siRNAs for ZIC2 and its control, total RNAs were purified and gene expression profiles were analyzed by AGILENT human 4x44k cDNA microarray. ZIC2 target genes were identified in PANC-1 cells. Gene expression profiles of PANC-1Tet/ZIC2 and PANC-1Tet/empty cells were analyzed by AGILENT human 4x44k cDNA microarray before and 48 hours after the DOX withdrawal. As well as ZIC2-inducible system, gene expression profiles of control- and ZIC2-knocdown PANC-1 cells were also analyzed by AGILENT human 4x44k cDNA microarray.

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The identification of auto-antibodies in pancreatic cancer patient sera using a naturally fractionated Panc-1 cell line.

Project description:The immunogenic nature of cancer can be explored to distinguish pancreatic cancer from related non-cancer conditions. We describe a liquid-based microarray approach followed by statistical analysis and confirmation for discovery of auto-immune biomarkers for pancreatic cancer. Proteins from the Panc-1 pancreatic cancer cell line were fractionated using a 2-D liquid separation method into over 1052 fractions and spotted onto nitrocellulose coated glass slides. The slides were hybridized with 37 pancreatic cancer sera, 24 chronic pancreatitis sera and 23 normal sera to detect elevated levels of reactivity against the proteins in spotted fractions. The response data obtained from protein microarrays was first analyzed by Wilcoxon Rank-Sum Tests to generate two lists of fractions that positively responded to the cancer sera and showed p-values less than 0.02 in the pairwise comparison between cancer specimens and normal and chronic pancreatitis specimens. The top 3 fractions with the lowest correlations were combined in receiver operating characteristic analyses. The area-under-the-curve (AUC) values are 0.813 and 0.792 for cancer vs. normal and cancer vs. pancreatitis respectively. Outlier-Sum statistics were then applied to the microarray data to determine the existence of outliers exclusive in cancer sera. The selected fractions were identified by LC-MS/MS. We further confirmed the occurrence of outliers with three proteins among cancer samples in a confirmation experiment using a separate dataset of 165 serum samples containing 48 cancer sera and 117 non-cancer controls. Phosphoglycerate kinase 1 (PGK1) elicited greater reactivity in 20.9% (10 in 48) of the samples in the cancer group, while no outlier was present in the non-cancer groups.

| S-EPMC3130890 | biostudies-literature

Metabolic characterization of invaded cells of the pancreatic cancer cell line, PANC-1.

Project description:We previously reported that about 0.4% of cells in the cultured human pancreatic cancer cell line, PANC-1, can invade matrigel during the transwell invasion assay, suggesting that these invaded PANC-1 cells may have specific characteristics to keep their invasive potential. To identify the metabolic characterization specific in the invaded PANC-1 cells, metabolome analysis of the invaded PANC-1 compared with the whole cultured PANC-1 was performed using CE-TOFMS, and concentrations of 110 metabolites were measured. In contrast to the whole cultured cells, the invaded PANC-1 was characterized as a population with reduced levels of amino acids and TCA cycle intermediates, and decreased and increased intermediates in glycolysis and nucleic acid metabolism. In particular, the ratio of both adenosine and guanosine energy charge was reduced in the invaded cells, revealing that the consumption of ATP and GTP was high in the invaded cells, and thus suggesting that ATP- or GTP-generating pathways are stimulated. In addition, the GSH/GSSG ratio was low in the invaded cells, but these cells had a higher surviving fraction after exposure to hydrogen peroxide. Thus, the invaded cells were the population resistant to oxidative stress. Furthermore, reduction in intracellular GSH content inhibited PANC-1 invasiveness, indicated that GSH has an important role in PANC-1 invasiveness. Overall, we propose the invaded cells have several unique metabolic profiles.

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Expression analysis of human pancreatic cancer cell lines (Capan-1, Panc-1, Panc-1+ve and Panc-1-ve) and pancreatic ductal cell line (HPDE)

Project description:Analysis of differentially expressing genes in whole genome wide analysis of aptamer SQ2 positive cells (Capan-1, Panc-1, Panc-1+ve) and SQ2 negative cells (Panc-1-ve and HPDE) Panc-1 +ve and Panc-1-ve cell lines were generated from Panc-1 cell line based upon its hetergenous binding to aptamer SQ2. Detailed procedure of generation of these cell lines are described in Pooja Dua, Hye Suk Kang, Seung-Mo Hong, Ming-Sound Tsao, Soyoun Kim, and Dong-ki Lee. 2012 Alkaline Phosphatase ALPPL2 is a novel pancreatic carcinoma-associated protein. Cancer Research A five chip study using total RNA recovered from Capan-1, Panc-1, Panc-1+ve, Panc-1-ve and HPDE cells. Each chip measures 45,033 genes with three 60 mer probe pairs per target.

| S-ECPF-GEOD-42428 | biostudies-other

Identification of RegIV as a novel GLI1 target gene in human pancreatic cancer.

Project description:GLI1 is the key transcriptional factor in the Hedgehog signaling pathway in pancreatic cancer. RegIV is associated with regeneration, and cell growth, survival, adhesion and resistance to apoptosis. We aimed to study RegIV expression in pancreatic cancer and its relationship to GLI1.GLI1 and RegIV expression were evaluated in tumor tissue and adjacent normal tissues of pancreatic cancer patients and 5 pancreatic cancer cell lines by qRT-PCR, Western blot, and immunohistochemistry (IHC), and the correlation between them. The GLI1-shRNA lentiviral vector was constructed and transfected into PANC-1, and lentiviral vector containing the GLI1 expression sequence was constructed and transfected into BxPC-3. GLI1 and RegIV expression were evaluated by qRT-PCR and Western blot. Finally we demonstrated RegIV to be the target of GLI1 by chromatin immunoprecipitation (CHIP) and electrophoretic mobility shift assays (EMSA).The results of IHC and qRT-PCR showed that RegIV and GLI1 expression was higher in pancreatic cancer tissues versus adjacent normal tissues (p<0.001). RegIV expression correlated with GLI1 expression in these tissues (R?=?0.795, p<0.0001). These results were verified for protein (R?=?0.939, p?=?0.018) and mRNA expression (R?=?0.959, p?=?0.011) in 5 pancreatic cancer cell lines. RegIV mRNA and protein expression was decreased (94.7±0.3%, 84.1±0.5%; respectively) when GLI1 was knocked down (82.1±3.2%, 76.7±2.2%; respectively) by the RNAi technique. GLI1 overexpression in mRNA and protein level (924.5±5.3%, 362.1±3.5%; respectively) induced RegIV overexpression (729.1±4.3%, 339.0±3.7%; respectively). Moreover, CHIP and EMSA assays showed GLI1 protein bound to RegIV promotor regions (GATCATCCA) in pancreatic cancer cells.GLI1 promotes RegIV transcription by binding to the RegIV gene promoter in pancreatic cancer.

| S-EPMC3073946 | biostudies-literature

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