BACKGROUND: Human breast cancer cell (BCC) lines are used extensively in biomedical research and are classified as estrogen receptor (ER)-positive or ER-negative. We used flow cytometry (FCM), reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting (WB) to assess ER expression in human BCC lines reported as being ER-positive (MCF7, T-47D, ZR-75-1) or ER-negative (MDA-MB-231, SK-BR-3, MDA-MB-453, HCC1954) to determine the validity of this classification. MATERIALS AND METHODS: ER was assessed in permeablized, fixed cells by FCM using two monoclonal anti-ERα antibodies and a polyclonal anti-ERβ antibody, in parallel with RT-PCR and WB. RESULTS: All of the cell lines expressed ERα and ERβ. Indirect immunofluorescence indicated that it was membrane and cytoplasmic ER that was being detected by FCM. Down-regulation by fulvestrant confirmed it was ER. CONCLUSION: These results demonstrate the importance of reassessing the ER status of human BCC lines that are used widely in biomedical research.
BACKGROUND:Humanbreast cancer cell (BCC) lines are used extensively in biomedical research and are classified as estrogen receptor (ER)-positive or ER-negative. We used flow cytometry (FCM), reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting (WB) to assess ER expression in human BCC lines reported as being ER-positive (MCF7, T-47D, ZR-75-1) or ER-negative (MDA-MB-231, SK-BR-3, MDA-MB-453, HCC1954) to determine the validity of this classification. MATERIALS AND METHODS:ER was assessed in permeablized, fixed cells by FCM using two monoclonal anti-ERα antibodies and a polyclonal anti-ERβ antibody, in parallel with RT-PCR and WB. RESULTS: All of the cell lines expressed ERα and ERβ. Indirect immunofluorescence indicated that it was membrane and cytoplasmic ER that was being detected by FCM. Down-regulation by fulvestrant confirmed it was ER. CONCLUSION: These results demonstrate the importance of reassessing the ER status of human BCC lines that are used widely in biomedical research.
Authors: Arvind Arora; Swetha Parvathaneni; Mohammed A Aleskandarany; Devika Agarwal; Reem Ali; Tarek Abdel-Fatah; Andrew R Green; Graham R Ball; Emad A Rakha; Ian O Ellis; Sudha Sharma; Srinivasan Madhusudan Journal: Mol Cancer Ther Date: 2016-11-11 Impact factor: 6.261
Authors: María F Montenegro; María Del Mar Collado-González; María Piedad Fernández-Pérez; Manel B Hammouda; Lana Tolordava; Mariam Gamkrelidze; José Neptuno Rodríguez-López Journal: BMC Cancer Date: 2014-07-26 Impact factor: 4.430