EGF-R antisense RNA blocks expression of the epidermal growth factor receptor and suppresses the transforming phenotype of a human carcinoma cell line

Maria Cristina Moroni, Mark C. Willingham, Laura Beguinot

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

We have used an antisense approach to investigate the role of overexpression of the normal human epidermal growth factor (EGF) receptor in the transformed phenotype of KB cells, which are a tumor derived human cell line. Initial experiments performed in vitro, showed that antisense RNA complementary to the entire coding region (AS-FL) or to parts of the EGF-R mRNA (AS-3', AS-5', and AS-K) effectively blocked translation of EGF-R mRNA. In addition, upon microinjection into KB cells, the in vitro synthesized antisense RNAs were able to inhibit transiently the synthesis of EGF-R. Inhibition was concentration-dependent, both in vitro and in cells, and the most effective constructs were those complementary to the entire coding region (ASFL) or to the 3'-coding end of the mRNA (AS-3'). Transfection of the same EGF-R antisense RNA constructs into the human epidermoid carcinoma KB cell line gave rise to several clones stably expressing elevated levels of antisense RNA and resulting in low residual levels of EGF receptor. The most reduced clones exhibited a totally restored serum-dependent growth and were severely impaired in colony formation and growth in agar. In addition the severity of the phenotype was directly proportional to the residual amount of EGF-R expressed. We conclude that overexpression of normal EGF-R plays a direct primary role in the development of the transformed phenotype of this human cancer cell line.

Original languageEnglish
Pages (from-to)2714-2722
Number of pages9
JournalJournal of Biological Chemistry
Volume267
Issue number4
Publication statusPublished - Feb 5 1992

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Antisense RNA
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Cells
Carcinoma
Phenotype
KB Cells
Cell Line
Messenger RNA
Clone Cells
Microinjections
Growth
Agar
Transfection
Tumors
Squamous Cell Carcinoma
Neoplasms
Serum
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

EGF-R antisense RNA blocks expression of the epidermal growth factor receptor and suppresses the transforming phenotype of a human carcinoma cell line. / Moroni, Maria Cristina; Willingham, Mark C.; Beguinot, Laura.

In: Journal of Biological Chemistry, Vol. 267, No. 4, 05.02.1992, p. 2714-2722.

Research output: Contribution to journalArticle

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