In situ amplification of oestrogen receptor alpha mRNA in breast cancer cell lines and tumours.
The aim of this work was to develop a direct in situ reverse transcription polymerase chain reaction (in situ RT-PCR) assay for the detection of oestrogen receptor alpha (ERalpha) mRNA on in vitro cell lines and breast tumour cell smears. ERalpha mRNA amplification was performed on MCF-7 (ERalpha positive) and MDA-MB-231 (ERalpha negative) cell lines as well as on 51 cytological smears of breast tumour samples from patients. The in situ amplification of mRNA in cell lines and ex vivo breast tumours was successful. However, finding an equilibrium between optimal cell morphology and PCR performance varied with each tumour, leading to difficulty in standardisation for daily practice. Nonetheless, in situ RT-PCR is a useful tool for the detection of ERalpha mRNA in selected cases, both in vitro and ex vivo. J Clin PATHOL: Mol Pathol
Lespagnard
L
L
Department of Pathology, Institut Jules Bordet, 1 rue Héger-Bordet, 1000 Brussels, Belgium. llespa@hotmail.com
Gancberg
D
D
Maaroufi
Y
Y
Leclercq
G
G
Larsimont
D
D
Verhest
A
A
D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism; D005260:Female; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis*; D011960:Receptors, Estrogen / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016707:Tissue Fixation / Q000379:methods; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D011960:Receptors, Estrogen