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Wolverhampton Intellectual Repository and E-Theses > Research Institutes > Research Institute in Healthcare Science > Cancer Research Group > Messenger RNA expression profiling of genes involved in epidermal growth factor receptor signalling in human cancer cells treated with scanning array-designed antisense oligonucleotides.

Please use this identifier to cite or link to this item: http://hdl.handle.net/2436/29816
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Title: Messenger RNA expression profiling of genes involved in epidermal growth factor receptor signalling in human cancer cells treated with scanning array-designed antisense oligonucleotides.
Authors: Petch, Amelia K.
Sohail, Muhammad
Hughes, Marcus D.
Benter, Ibrahim
Darling, John L.
Southern, Edwin M.
Akhtar, Saghir
Citation: Biochemical Pharmacology, 66(5): 819-830
Publisher: Amsterdam: Elsevier
Journal: Biochemical Pharmacology
Issue Date: 2003
URI: http://hdl.handle.net/2436/29816
DOI: 10.1016/S0006-2952(03)00407-6
PubMed ID: 12948863
Additional Links: http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T4P-49621XH-2&_user=1644469&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000054077&_version=1&_urlVersion=0&_userid=1644469&md5=193199e877eb410bf0ca9f0c128a8297
Abstract: Scanning oligodeoxynucleotide (ODN) arrays appear promising in vitro tools for the prediction of effective antisense reagents but their usefulness has not yet been reported in mammalian systems. In this study, we have evaluated the use of scanning ODN arrays to predict efficacious antisense ODNs targeting the human epidermal growth factor receptor (EGFR) mRNA in a human epidermoid cancer cell line and in primary human glioma cells. Hybridisation accessibility profile of the first 120nt in the coding region of the human EGFR mRNA was determined by hybridising a radiolabelled EGFR transcript to a scanning array of 2684 antisense sequences ranging from monomers to 27-mers. Two ODNs, AS1 and AS2, complementary to accessible sequences within the EGFR mRNA, were designed and their ability to hybridise to EGFR mRNA was further confirmed by in vitro RNase H-mediated cleavage assays. Phosphorothioate-modified 21-mer AS1 and AS2 ODNs inhibited the growth of an established human A431 cancer cell line as well as primary glioma cells from human subjects when delivered as cationic lipoplexes. In contrast, scrambled controls and AS3-an antisense ODN complementary to an inaccessible site in EGFR mRNA-were inactive. Western blots showed that AS1 ODN exhibited a dose-dependent inhibition of EGFR protein expression in A431 cells in the nanomolar range. Microarray-based gene expression profiling studies of A431 cells treated with the 21-mer phosphorothioate AS1 ODN demonstrated successful inhibition of downstream signalling molecules further confirming the effective inhibition of EGFR expression in human cancer cells by antisense ODNs designed by scanning ODN array technology.
Type: Article
Language: en
Keywords: Oncology
A431 cells
Cell Culture
Epidermal Growth Factor Receptor (EGFR)
Oligodeoxynucleotide
Phosphorothioate
Skin cells
In vitro cultivation
Expression profiling
Primary tumour
MeSH: Cell Division
Down-Regulation
Gene Expression Profiling
Gene Expression Regulation, Neoplastic
Glioma
Humans
Oligonucleotide Array Sequence Analysis
Oligonucleotides, Antisense
RNA, Messenger
Receptor, Epidermal Growth Factor
Ribonuclease H
Signal Transduction
Tumor Cells, Cultured
Cell Line, Tumor
Cell Culture
ISSN: 0006-2952
Appears in Collections: Cancer Research Group

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