Ca 125

The deparaffinized sections of the original tumor and heterotransplanted tumor on glass slides were stained immunohistochemically using the universal Immuno-enzyme Polymer (UIP) method (HISTOFINE simple stain MAX PO; Nichirei Bioscience Inc., Tokyo, Japan). They were immersed in 0.03 % hydrogen peroxidase and absolute methanol to block endogenous peroxidase. The samples were then washed with phosphate buffered saline and heated in a microwave oven for 10 min at a power of 700 W. After cooling, the slides were incubated with a primary antibody at 4 °C for 12 h and then added and reacted with Histfine Simple stain MAX PO at room temperature for 30 min, followed by diaminobenzidine (DAB) for sections for 5–10 min. Antibodies used as tumor markers were CA 125 (DAKO, Glostrup, Denmark), CA 19–9 (DAKO) and, as hormone receptor markers, were anti-human estrogen receptor α (Nichirei) and anti-human progesterone receptor (Nichirei). Hepatocyte factor-1β (HNF-1β) [8 (link)] (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and Annexin IV [9 (link)] (Santa Cruz Biotech) which commonly express in ovarian clear cell carcinoma were also used.