Real-Time PCR: Detection of Oestrogen Receptor-Alpha and -Beta Isoforms and Variants in Breast Cancer

• Published in: Anticancer research 2010-10, Vol.30 (10), p.4147-4156
• Main Authors: AL-BADER, Maie, AL-SAJI, Shorooq, FORD, Christopher H. J, FRANCIS, Issam, AL-AYADHY, Bushra
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Cell Line, Tumor; Estrogen Receptor alpha - biosynthesis; Estrogen Receptor alpha - genetics; Estrogen Receptor beta - biosynthesis; Estrogen Receptor beta - genetics; Female; Gene Amplification; Gynecology. Andrology. Obstetrics; Humans; Mammary gland diseases; Medical sciences; Paraffin Embedding; Reverse Transcriptase Polymerase Chain Reaction - methods; Tumors
• ISSN: 0250-7005; 1791-7530

Splice variants exist for both alpha and beta oestrogen receptors (ERs). Oestrogen function results from a balance between the wild-type ERs (wt) and their variants. Forty formalin-fixed paraffin-embedded breast cancer samples were analysed by real-time PCR using ERα primer sets detecting wt and exon-deleted 3, 5, 6 and 7 variants. The ERβ primer sets detected wt ERβ1 and ERβ2 and ERβ5 variants. At the end of the PCR cycles, a dissociation curve was generated showing the peaks for each sample at specific melting temperatures (Tm); finding more than one peak indicated the presence of variants. Many samples expressed both wt ER isoforms and their variants. The Tm value served as a cut-off point for determination of wt versus variant ER expression. This method of detection of wt and variant ER could help in patient selection for anti-oestrogen therapy and in monitoring response to therapy.