Biomic study of human myeloid leukemia cells differentiation to macrophages using DNA array, proteomic, and bioinformatic analytical methods

Juan, Hsueh‐Fen; Lin, John Yi-Chung; Chang, Wen-Hwei; Wu, Ching‐Yuan; Pan, Tai‐Long; Tseng, Min‐Jen; Khoo, Kay‐Hooi; Chen, Shui‐Tein

doi:10.1002/1522-2683(200208)23:15<2490::aid-elps2490>3.0.co;2-3

Cited by 59 publications

(28 citation statements)

References 73 publications

(62 reference statements)

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“…The labeled targets were hybridized on commercial 7.6 K cDNA microarrays representing approximately 7500 distinct human transcripts (Egenomix Human UniversoChip 8K; Egenomix Technology Corporation, Taipei) with detail description reported earlier (Juan et al, 2002).…”

Section: Mmp-2 Activity Assaymentioning

confidence: 99%

Insulin-like growth factor binding protein-3 (IGFBP-3) acts as an invasion-metastasis suppressor in ovarian endometrioid carcinoma

et al. 2007

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Metastasis and invasion occur in the majority of epithelial ovarian carcinoma at diagnosis. To delineate the molecular signature in ovarian cancer invasion, we established and characterized a human ovarian endometrioid carcinoma (EC) cell line OVTW59-P0 and its invasion-related sublines (P1-P4, in the order of increasing invasive activity). P4 showed faster migration and larger xenograft formation with metastasis than P0. By microarray analysis of different gene expression among P0-P4 sublines, one group of gene was found negatively correlated with cancer invasion. Among these genes, IGFBP-3 was identified as one of the most remarkably suppressed gene that showed lower gene expression in P4 than P0. Re-expression of IGFBP-3 in P4 effectively inhibited cell migration, invasion and metastasis, but did not affect cell proliferation. In 35 patients with EC tumors, low IGFBP-3 expression correlated clinically with higher tumor grade, advanced stage and poor survival. Our results provide evidence and indicate that IGFBP-3 plays an important role as an invasion-metastasis suppressor in ovarian EC.

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Section: Mmp-2 Activity Assaymentioning

confidence: 99%

Insulin-like growth factor binding protein-3 (IGFBP-3) acts as an invasion-metastasis suppressor in ovarian endometrioid carcinoma

et al. 2007

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“…Two-dimensional gel electrophoresis (2DE) combined with protein identification by mass spectrometry has already defined differences in hematopoietic cell lines which occur during differentiation or as a result of BCR/ABL oncogene expression. [4][5][6][7] These approaches relied on intergel sample comparison using silver or Coomassie blue stains, which have a low dynamic range and thus could be subject to the inherent limitations of this classical experimental approach. The development of protein labeling techniques employing fluorescent dyes with distinct excitation/ emission spectra but with the same chemical reactivity 8,9 enables multiplexing of 2 or more samples so that they can be processed and run on the same gel.…”

Section: Introductionmentioning

confidence: 99%

Comparative proteomics of primitive hematopoietic cell populations reveals differences in expression of proteins regulating motility

Evans

Tonge²,

Blinco³

et al. 2004

Blood

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Lineage-marker depleted (Lin ؊ ) murine bone marrow cells expressing stem cell antigen 1 (Sca-1) were sorted on the basis of stem cell factor receptor (c-kit) expression to obtain Lin ؊ Sca ؉ Kit ؉ or Lin ؊ Sca ؉ Kit ؊ cells. Lin ؊ Sca ؉ Kit ؊ cells have a markedly greater chemotactic response to stromal derived factor-1 (SDF-1). Using a novel fluorescent stain, we show that both populations generate similar levels of a key messenger, phosphatidylinositol 3,4,5 trisphosphate (PIP 3 ), in response to SDF-1. Differences in motile behavior may therefore lie downstream of phosphatidylinositol 3-kinase (PI3-kinase) activation at the level of cytoskeleton regulation. The 2 cell populations were compared using 2-dimensional difference gel electrophoresis (2D-DIGE), with a maleimide CyDye fluorescent protein labeling technique that has enhanced sensitivity for low abundance samples. Comparative proteomic analysis of Cy3-and Cy5-labeled protein samples allows relative quantification of protein spots present in both cell populations; of these, 73% were common. Key protein differences were adseverin and gelsolin, actin microfilament splicing proteins, regulated by Rac, downstream of PI3-kinase activation. Adseverin was shown to be acetylated, a novel modification for this protein. Differences in major regulators of cell shape and motility between the 2 populations can explain the differential response to SDF-1. IntroductionHematopoietic stem cells have the capacity to self-renew and also to differentiate to form a number of mature cell types with distinct morphologies and functional activities. The distinctive features of the stem cell that give it longevity and the ability to balance its self-renewal and differentiation to the required degree in any circumstance remain unclear. These biologic properties are encoded in the transcriptional profile, or transcriptome, of stem cells. The properties of a stem cell can therefore be defined in terms of the gene expression profiles of stem cells. Retention of stem cell characteristics will rely in part on the cohort of genes specifically expressed in these primitive cells, but not in their maturing progeny. This axiom has led to transcriptional analysis of long-and short-term repopulating stem cells and other primitive populations (defined via expression of specific markers) to define the genes expressed for self-renewal and pluripotency. 1,2 The generation of these stem cell "blueprints" then offers the opportunity to investigate the functional activity of the proteins produced as a consequence of gene expression and to systematically study gene networks controlling stem cell biology.The relationship between messenger RNA production and protein synthesis is not, however, strictly correlative. 3 In a range of different organisms it has become clear that changes in the expression of a specific protein do not necessarily relate directly to altered transcription. Recent developments in protein separation and relative quantification allow populations of cells to be compared at the pr...

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“…After sonication, 500 mg of total protein was loaded into immobilized pH gradient (IPG) gel strips (pH 3-10 or pH 4-7, 18 cm long; Amersham Pharmacia Biotech, Uppsala, Sweden). For the first-dimensional separation, isoelectric focusing was carried out using the IPGphor system (Amersham Pharmacia Biotech) at 20jC with 8000 V for a total of 65 kVh as previously described (15). After isoelectric focusing, the IPG strips were equilibrated for 15 minutes then attached with 0.5% agarose to the top of a vertical 12.5% SDSpolyacrylamide gel.…”

Section: Methodsmentioning

confidence: 99%

“…After isoelectric focusing, the IPG strips were equilibrated for 15 minutes then attached with 0.5% agarose to the top of a vertical 12.5% SDSpolyacrylamide gel. Second-dimensional electrophoresis was carried out with Protean II Multi-Cell (Bio-Rad, Hercules, CA) as previously described (15). The gels were fixed and stained in 350 mL of the Sypro Ruby Protein Gel (Invitrogen, Carlsbad, CA) stain solution overnight, then the residual dye was washed out of the polyacrylamide matrix.…”

Section: Methodsmentioning

confidence: 99%

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Cytotoxicity and Proteomics Analyses of OSU03013 in Lung Cancer

Tan¹,

Lee²,

Lin

et al. 2008

Clinical Cancer Research

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Purpose: Most lung cancer patients have some resistance to and suffer from side effects of conventional chemotherapy. Thus, identification of a novel anticancer drug with better target selectivity for lung cancer treatment is urgently needed. Experimental Design: In order to investigate whether OSU03013, a derivative of celecoxib, can be a potential drug for lung cancer treatment, we examined its cytotoxicity mechanisms by flow cytometry and phosphatidylserine staining in A549, CL1-1, and H1435 lung cancer cell lines, which are resistant to the conventional drug, cisplatin. In addition, we identified the affected proteins by proteomics and confirmed the selected proteins by Western blot analysis. We examined the interaction between OSU03013 and potential target protein by molecular modeling. Results: Our results indicated that OSU03013 had low-dose (1f4 AM) cytotoxicity in all lung cancer cell lines tested 48 hours posttreatment. OSU03013 caused cell cycle G1phase arrest and showed phosphatidylserine early apoptosis via endoplasmic reticulum stress. Several proteins such as heat shock protein 27, 70, and 90, CDC2, a-tubulin, annexin A3, cAMP-dependent protein kinase, glycogen synthase kinase 3-beta, and h-catenin were identified by proteomics and confirmed by Western blot. In addition, molecular modeling showed that OSU03013 competes with ATP to bind to cAMP-dependent protein kinase. Conclusions: We identified for the first time that OSU03013 inhibits cAMP-dependent protein kinase activity and causes dephosphorylation of glycogen synthase kinase 3-beta leading to h-catenin degradation, which is often overexpressed in lung cancer. Our molecular and proteomic results show the potential of OSU03013 as an anticancer drug for lung cancer.

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Biomic study of human myeloid leukemia cells differentiation to macrophages using DNA array, proteomic, and bioinformatic analytical methods

Cited by 59 publications

References 73 publications

Insulin-like growth factor binding protein-3 (IGFBP-3) acts as an invasion-metastasis suppressor in ovarian endometrioid carcinoma

Insulin-like growth factor binding protein-3 (IGFBP-3) acts as an invasion-metastasis suppressor in ovarian endometrioid carcinoma

Comparative proteomics of primitive hematopoietic cell populations reveals differences in expression of proteins regulating motility

Cytotoxicity and Proteomics Analyses of OSU03013 in Lung Cancer

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