This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chen, Z.-P. Articles by Panasci, L. C. Search for Related Content PubMed PubMed Citation Articles by Chen, Z.-P. Articles by Panasci, L. C.

Correlation of Chloroethylnitrosourea Resistance with ERCC-2 Expression in Human Tumor Cell Lines as Determined by Quantitative Competitive Polymerase Chain Reaction¹

Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, 3755 Côte Ste. Catherine Road, Montreal, Quebec, H3T 1E2 Canada

We have developed a method to quantitate ERCC-2 gene expression in tumor cell lines. A mutant ERCC-2 DNA fragment (1-bp mutation) is used as a competitive DNA template in a coamplification PCR reaction with cDNA obtained by reverse transcribing DNase-free total RNA from six human tumor cell lines. The PCR products are separated on agarose gel by virtue of their differential banding pattern upon restriction enzyme digestion. Densitometric readings of the PCR products from a negative film of the gel are used to establish a linear regression curve, which in turn is used to quantitate ERCC-2 levels. ß-actin expression is similarly quantitated. Normalized ERCC-2 gene expression (either to ß-actin or to total RNA) correlates with cytotoxicity of 1,3-bis-(2-chloroethyl)-1-nitrosourea or (2-chloroethyl)-3-sarcosinamide-1-nitrosourea, suggesting that ERCC-2 may play an important role in drug resistance in these cell lines. This method is reliable and can be used to quantitate gene expression in clinical tumor specimens.

¹ This work is supported by the National Institute of Neurological Disorders and Stroke Grant NS22230 and by a private donation from Helen and Nicki Lang.

² To whom requests for reprints should be addressed.

Received 1/17/96. Accepted 4/17/96.

This article has been cited by other articles:

				Z.-P. Chen, J. Remack, T. P. Brent, G. Mohr, and L. C. Panasci Extraneuronal Monoamine Transporter Expression and DNA Repair Vis-a-Vis 2-Chloroethyl-3-sarcosinamide-1-nitrosourea Cytotoxicity in Human Tumor Cell Lines Clin. Cancer Res., December 1, 1999; 5(12): 4186 - 4190. [Abstract] [Full Text] [PDF]

				Z.-P. Chen, A. Malapetsa, A. McQuillan, D. Marcantonio, V. Bello, G. Mohr, J. Remack, T. P. Brent, and L. C. Panasci Evidence for Nucleotide Excision Repair as a Modifying Factor of O6-Methylguanine-DNA Methyltransferase-Mediated Innate Chloroethylnitrosourea Resistance in Human Tumor Cell Lines Mol. Pharmacol., November 1, 1997; 52(5): 815 - 820. [Abstract] [Full Text]