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Title: Induction of marked apoptosis in mammalian cancer cell lines by antisense DNA treatment to abolish expression of DENN (Differentially Expressed in Normal and Neoplastic Cells)
Authors: Lim, K.M. 
Chow, V.T.K. 
Keywords: Antisense
Death domain
Issue Date: 1-Nov-2002
Source: Lim, K.M., Chow, V.T.K. (2002-11-01). Induction of marked apoptosis in mammalian cancer cell lines by antisense DNA treatment to abolish expression of DENN (Differentially Expressed in Normal and Neoplastic Cells). Molecular Carcinogenesis 35 (3) : 110-126. ScholarBank@NUS Repository.
Abstract: We previously reported the isolation of the novel human DENN gene, which is differentially expressed in normal and neoplastic cells. DENN is identical to MADD (mitogen-activated protein kinase-activating death domain), which interacts with tumor necrosis factor receptor 1 through their death domains. DENN is also homologous to Rab3 GEP, a rat Rab3 GDP/GTP exchange protein. Real-time reverse transcription -polymerase chain reaction analysis showed that DENN expression in cancer cell lines was 26-50 times that in normal cells. The Jurkat human leukemia, PLC/PRF/5 human hepatoma, and NS-1 mouse myeloma cell lines as well as the MRC-5 human fetal lung and Vero monkey kidney cell lines were treated successfully with four separate DENN-targeted antisense oligodeoxynucleotides (ODNs) to abrogate DENN expression. Quantitative assessment of cell viability and apoptosis by flow cytometry via fluorescein diacetate and propidium iodide membrane-integrity tests, terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate-biotin nick end-labeling, and annexin V assays showed that antisense silencing of DENN resulted in markedly more pronounced cell death in cancer cells compared with nonmalignant cells. Antisense-treated cell lines exhibited extensive loss of DNA content, forming distinct sub-G1 peaks, while cell proliferation diminished significantly. Ultrastructural features of programmed cell death in cells subjected to antisense ODNs were authenticated by electron microscopy. In contrast, transfection of cell lines with a plasmid construct to achieve DENN overexpression augmented cellular proliferation and could reverse the apoptotic effect of antisense and staurosporine treatment. Our findings suggest that DENN is intimately involved in anti-apoptotic and cell-survival processes. © 2002 Wiley-Liss, Inc.
Source Title: Molecular Carcinogenesis
ISSN: 08991987
DOI: 10.1002/mc.10082
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