Abstract
Competitive polymerase chain reaction assays have been developed for the quantitation of oestrogen receptor mRNA and two oestrogen-regulated mRNAs (progesterone receptor and pNR-2/pS2) in breast cancer cells. These assays are more sensitive than traditional hybridisation techniques, do not require the use of radioisotopes, measure absolute amounts of messenger RNAs and can be used to measure the expression of mRNAs in small numbers of tumour cells obtained by fine-needle aspiration (FNA). These assays should prove useful for predicting the hormone responsiveness of breast cancer from tumour cells obtained by FNA at diagnosis and could be particularly useful in the management of elderly/frail patients who receive primary tamoxifen, or in other patients for whom tumour tissue for standard biochemical measurements is not available.
Highlights
Breast cancer is frequently oestrogen responsive and this has resulted in the widespread use of hormonal therapy in its treatment
In this report we describe the development of competitive reverse transcription-polymerase chain reaction (RT-PCR) assays based on the procedure described by Becker-Andre and Hahlbrock (1989), for the measurement of oestrogen receptor, progesterone receptor and pNR-2/pS2 mRNA levels in RNA extracted from small numbers of breast cancer cells
The competitor RNA differs from the normal cellular RNA being measured by a single nucleotide and the single nucleotide difference is sufficient to introduce a restriction enzyme site into the centre of PCR products derived from the competitor RNA
Summary
The aim of this study was to develop competitive RT-PCR assays that would permit measurement of the target mRNAs in the small numbers of tumour cells that are obtained by FNA
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