TY - JOUR
T1 - Re-evaluation of the prolactin receptor expression in human breast cancer
AU - Galsgaard, Elisabeth Douglas
AU - Rasmussen, Birgitte Bruun
AU - Folkesson, Charlotta Grånäs
AU - Rasmussen, Louise Maymann
AU - Berchtold, Martin Werner
AU - Christensen, Leif
AU - Panina, Svetlana
N1 - Keywords: Animals; Antibody Specificity; Breast Neoplasms; CHO Cells; Carcinoma; Cricetinae; Cricetulus; Female; Gene Expression Regulation, Neoplastic; HT29 Cells; Humans; Prolactin; Protein Binding; Receptors, Prolactin; Tissue Array Analysis; Tumor Cells, Cultured
PY - 2009
Y1 - 2009
N2 - The pituitary hormone PRL is involved in tumorigenesis in rodents and humans. PRL promotes proliferation, survival and migration of cancer cells acting via the PRL receptor (PRLR). Aiming to perform a large-scale immunohistochemical (IHC) screening of human mammary carcinomas for PRLR expression, we evaluated the specificity of commercially available anti-human PRLR antibodies (B6.2, U5, PRLRi pAb, 1A2B1, 250448 and H-300). The latter three antibodies were found to specifically recognise PRLR. The relative PRLR expression level detected with these antibodies closely reflected the level of (125)I-PRL binding to the cell surface. The monoclonal antibody (mAb) 250448 was specific for the N-()glycosylated form of PRLR and blocked PRL binding and signalling. The PRLRi polyclonal antibody recognised cytokeratin-18. The mAb B6.2, previously used in a number of studies, was found to lack specificity for PRLR and to rather recognise a PRLR-associated protein. The mAb U5 raised against the rat PRLR did not cross-react with the human receptor. Only one mAb, 1A2B1, was found useful for detection of PRLR in IHC applications. This antibody recognised PRLR expressed in human breast cancer cell lines and decidual cells in tissue sections of human placenta. Screening of 160 mammary adenocarcinomas demonstrated significant immunoreactivity in only four tumours, indicating that PRLR is generally not strongly upregulated in human breast cancer. However, even a very low level of PRLR expression was found to be sufficient to mediate PRL responsiveness in breast cancer cell lines.
AB - The pituitary hormone PRL is involved in tumorigenesis in rodents and humans. PRL promotes proliferation, survival and migration of cancer cells acting via the PRL receptor (PRLR). Aiming to perform a large-scale immunohistochemical (IHC) screening of human mammary carcinomas for PRLR expression, we evaluated the specificity of commercially available anti-human PRLR antibodies (B6.2, U5, PRLRi pAb, 1A2B1, 250448 and H-300). The latter three antibodies were found to specifically recognise PRLR. The relative PRLR expression level detected with these antibodies closely reflected the level of (125)I-PRL binding to the cell surface. The monoclonal antibody (mAb) 250448 was specific for the N-()glycosylated form of PRLR and blocked PRL binding and signalling. The PRLRi polyclonal antibody recognised cytokeratin-18. The mAb B6.2, previously used in a number of studies, was found to lack specificity for PRLR and to rather recognise a PRLR-associated protein. The mAb U5 raised against the rat PRLR did not cross-react with the human receptor. Only one mAb, 1A2B1, was found useful for detection of PRLR in IHC applications. This antibody recognised PRLR expressed in human breast cancer cell lines and decidual cells in tissue sections of human placenta. Screening of 160 mammary adenocarcinomas demonstrated significant immunoreactivity in only four tumours, indicating that PRLR is generally not strongly upregulated in human breast cancer. However, even a very low level of PRLR expression was found to be sufficient to mediate PRL responsiveness in breast cancer cell lines.
U2 - 10.1677/JOE-08-0479
DO - 10.1677/JOE-08-0479
M3 - Journal article
C2 - 19153125
SN - 0022-0795
VL - 201
SP - 115
EP - 128
JO - Journal of Endocrinology
JF - Journal of Endocrinology
IS - 1
ER -