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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 2000 Apr;91(4):399–409. doi: 10.1111/j.1349-7006.2000.tb00959.x

Screening of BRCA1 Mutation Using Immunohistochemical Staining with C‐Terminal and N‐Terminal Antibodies in Familial Ovarian Cancers

Katsunori Kashima 1, Takashi Oite 2, Yoichi Aoki 1, Koichi Takakuwa 1, Hiroshi Aida 1, Hiroshi Nagata 1, Masayuki Sekine 1, Hong Jun Wu 1, Yasuo Hirai 3, Yuichi Wada 4, Kaichiro Yamamoto 5, Kazuo Hasegawa 6, Takahiko Sonoda 7, Takeshi Maruo 8, Ichiro Nagata 9, Masayuki Ohno 10, Mitsuaki Suzuki 11, Iwao Kobayashi 12, Kazuo Kuzuya 13, Takeshi Takahashi 14, Yuichi Torii 15, Kenichi Tanaka 1,✉
PMCID: PMC5926460  PMID: 10804288

Abstract

We examined the subcellular localization of BRCA1 proteins using immunohistochemical staining with C‐terminal (GLK‐2 antibody) and N‐terminal (Ab‐2 antibody) monoclonal antibodies in 44 familial ovarian cancers. Among these, 24 cases were associated with 13 independent germ‐line mutations of BRCA1, and loss of heterozygosity (LOH) at one or more BRCA1 microsatellite markers was found in all 21 informative tumors tested. With GLK‐2 antibody, cytoplasmic staining was observed in 15 of 16 tumors (93.8%) with mutation in exon 11, and BRCA1 staining was absent in 8 of 8 tumors (100%) with mutation in exons other than exon 11. When immunohistochemical staining was performed with Ab‐2 antibody, both nuclear and cytoplasmic staining were observed in 14 of 16 tumors (87.5%) with mutation in exon 11. Interestingly, nuclear staining was observed in 3 of 3 tumors (100%) with mutation downstream of exon 11, even though no staining was detected in 5 of 5 tumors (100%) with mutation upstream of exon 11. On the other hand, in familial ovarian cancers without BRCA1 mutations, nuclear staining or both nuclear and cytoplasmic staining was observed in 18 of 20 specimens (90%) and 20 of 20 specimens (100%) with GLK‐2 antibody and with Ab‐2 antibody, respectively. These results suggest that an immunohistochemical assay in combination with employing the C‐terminal and the N‐terminal antibodies appears to have potential as a reliable and useful technique for the screening of BRCA1 mutations, at least to predict the status of mutation, upstream or downstream of exon 11.

Keywords: Familial ovarian cancer, BRCA1, Subcellular localization, Splice variant, Immunohistochemistry

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