Fig 1.
SDS–PAGE separation and western blotting of purified aromatase.
Microsome pellets of human term placenta were solubilized and applied to an immunoadsorbent column coupled with a monoclonal antibody (MAb3-2C2) specific to aromatase. The eluted aromatase was subjected to SDS–PAGE and western blotting. A, SDS–PAGE separated fractions of the purified aromatase (stained with coomassie brilliant blue: CBB). B, western blot of aromatase (immunostained with R-8-1, a polyclonal antiserum specific to aromatase). C, negative control (reacted with normal rabbit serum). Lanes: 1, molecular weight markers; 2, the purified aromatase.
Fig 2.
Reactivity of antisera to aromatase.
The sensitivity and specificity of antisera to aromatase were evaluated by an ELISA using A, purified aromatase; B, a placental microsome extract; and C, a liver microsome extract, as immobilized antigens. BL, blank. Western blotting with antisera to aromatase for D, a placental microsome extract, and E, a liver microsome extract. The extracts were separated by SDS–PAGE and electrotransferred to a PVDF membrane. This was subjected to western blotting with each of three antisera (Rabbits 1, 2, and 3) using antiserum R-8-1[11–13] as the positive control. Lanes: 1, molecular weight markers; 2, CBB staining; 3, R-8-1; 4, normal rabbit serum (negative control); 5, Rabbit 1; 6, Rabbit 2; and 7, Rabbit 3.
Fig 3.
Immunohistochemistry of aromatase in breast cancer tissues.
A, strong aromatase expression of a case that was negative for ER, PgR and HER2, with histologic grade 2 and pT1bN0M0. B, weak aromatase expression of a case that was positive for ER and PgR; negative for HER2, with histologic grade 2 and pT1cN1aM0. C, negative aromatase expression of a case that was positive for ER and PgR; negative for HER2, with histologic grade 2 and pT1cN0M0. D, positive control from human term placenta. E, negative control from normal human liver. F, double immunostaining for ER (brown) and aromatase (red) in tissue that was positive for ER, PgR, and negative for HER2, with histologic grade 2 and pT1cN0M0. G, double immunostaining for PgR (brown) and aromatase (red) in tissue that was positive for ER, PgR and negative for HER2, with histologic grade 2 and pT1cN0M0. H, double immunostaining for HER2 (brown) and aromatase (red) in tissue that was negative for ER, PgR and positive for HER2, with histologic grade 2 and pT1cN3aM0.
Table 1.
Summary of clinicopathological factors.
Fig 4.
Immunohistochemistry of aromatase in normal breast tissue.
Table 2.
Correlation between aromatase expression level and various clinicopathological factors.