Simultaneous measurement of nucleolin and estrogen receptor in breast cancer cells by laser scanning cytometry.
BACKGROUND
The purpose of the study was to test the feasibility of laser scanning cytometry (LSC) to simultaneously measure estrogen receptor (ER) and nucleolin (NU) expression in the nuclei of the same individual breast cancer cells.


METHODS
Cancer cells from 64 breast tumors were labeled with anti-NU and biotinylated anti-ER antibodies, then with secondary FITC-conjugated antibody and streptavidin-APC conjugate, respectively, and measured by LSC. The expression of NU in the nucleus and NU aggregates (NUA), number of NUA, nuclear and NUA areas and ER expression were assessed for aspiration each cell.


RESULTS
ER-bound APC fluorescence correlated with nuclear NU (r=0.65; p<0.001) and NUA-bound FITC fluorescence (r=0.59; p<0.001). Good correlation was found between percentages of ER-positive cells in LSC and by image analysis in paraffin-embedded sections (r=0.59, p<0.001).


CONCLUSIONS
ER and NU expression can be measured simultaneously in the same nuclei of breast cancer cells.
Masiuk
Marek
M
Department of Pathology, Pomeranian Medical University, 71-252 Szczecin, Poland. mmasiuk@med.pam.szczecin.pl
Urasinska
Elzbieta
E
Domagala
Wenancjusz
W
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000737:chemistry*; D002467:Cell Nucleus / Q000737:chemistry; D005240:Feasibility Studies; D005260:Female; D005453:Fluorescence; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D008875:Middle Aged; D010750:Phosphoproteins / Q000032:analysis*; D016601:RNA-Binding Proteins / Q000032:analysis*; D011960:Receptors, Estrogen / Q000032:analysis*; D000097796:Nucleolin
D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D011960:Receptors, Estrogen; D000097796:Nucleolin