A quantitative assay for IGF-I and IGF binding protein mRNAs: expression in malignant melanoma cells

Molecular and Cellular Endocrinology
R C OlneyD M Wilson

Abstract

The quantification of messenger RNA is central in studies of gene expression. We describe a quantitative assay for specific mRNAs (QASM) that measures mRNAs for insulin-like growth factor-I, IGF binding proteins (IGFBPs) -2, -3, -4, and -5, and beta-actin. The assay utilizes reverse transcription and polymerase chain reaction, followed by an ELISA based DNA assay technique. The use of internal (competitive) quantification standards gave poorly linear results, while external standards gave linear and reproducible results. QASM results correlated with IGFBP protein concentrations in conditioned medium and with mRNA levels determined by Northern blotting. QASM was used to study IGFBP expression in human malignant melanoma cells. Messenger RNA for IGFBP-2, -3, and -5 were present, while IGF-I and IGFBP-4 mRNAs were not detected. IGFBP-2 and -3 expression was increased in a dose dependent manner by treatment with IGF-I. Protein concentrations in conditioned media paralleled mRNA levels. QASM is a sensitive, specific, and reproducible approach to determining mRNA levels.

References

Aug 1, 1990·Journal of Molecular Endocrinology·F Z UptonF J Ballard
Apr 1, 1991·The Journal of Clinical Endocrinology and Metabolism·F LiuR L Hintz
Jun 28, 1991·Biochemical and Biophysical Research Communications·S K SprattA Sommer
Jun 21, 1991·Science·H A ErlichJ J Sninsky
Apr 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·G GillilandH F Bunn
Dec 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·A M WangD F Mark
Sep 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·S Nakajima-IijimaT Kakunaga
Aug 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·E S KawasakiF P McCormick
Dec 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·W L LoweD LeRoith
Mar 25, 1983·Nucleic Acids Research·U NudelD Yaffe
Dec 8, 1983·Nature·M JansenJ L Van den Brande
Aug 27, 1993·Annals of the New York Academy of Sciences·D R ClemmonsG Wright
Aug 1, 1993·Current Opinion in Immunology·C S Subauste, J S Remington
Jan 1, 1993·Annual Review of Physiology·W S Cohick, D R Clemmons

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Citations

Dec 28, 1999·Molecular and Cellular Endocrinology·R C Olney, E B Mougey
Jan 28, 2004·Journal of Cutaneous Pathology·Huamin WangVictor G Prieto
Apr 20, 2004·Biochemical and Biophysical Research Communications·Robert C OlneyEdward B Mougey
Nov 3, 2005·American Journal of Physiology. Endocrinology and Metabolism·Prabhakaran BalagopalDavid Hammond
May 14, 1999·The Journal of Clinical Endocrinology and Metabolism·S R EdmondsonG A Werther
Dec 13, 2003·Endocrine Reviews·Stephanie R EdmondsonChristopher J Wraight

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