All of the cancer patients had no history or either chemotherapy or radiation therapy prior to the surgical staging. Family history of ovarian cancer and personal history of breast cancer were collected, but BRCA mutation status was not available. In addition to the tissue samples obtained
from the above HGSC patients, we also studied tubal tissues from a group of patients Luminespib with benign gynecologic diseases (n = 60) as negative controls. These patients had no evidence of any malignancy and came to the hospital for total hysterectomies and bilateral salpingo-oophorectomy because of leiomyomata, endometriosis, or uterine prolapse. The ages ranged from 42 to 75 with an average age of 61.5 years. Tissue handling All of the fallopian
tube samples Combretastatin A4 were handled using SEE-FIM protocol [3,25] for those cancer patients since this is the routine procedure in UMC. Fallopian tubes from benign control cases were processed by embedding all fimbriated ends similar to cancer patients with additional representative 2 cross sections of the ampulla as described previously [10]. All tissues were fixed in 10% buffered formalin and processed routinely for paraffin embedding. Five-micron sections for IHC were cut and placed on Super Plus slides (Fisher Scientific, Pittsburgh, PA) before sectioning each specimen for hematoxylin and eosin staining in order for them to be examined microscopically C59 chemical structure for diagnostic confirmation. Morphologic analysis The secretory and ciliated cells within the tubal mucosa were readily identifiable under the light microscopy. To further
confirm the cell type, we stained the tubal sections with PAX8 (marker for secretory cells) and tubulin (marker for ciliated cells). STIC is a noninvasive carcinoma confined to the epithelial cells of fimbriae and is characterized by significant cytologic atypia and/or atypical intraepithelial proliferation. The histologic diagnoses of STIC were made based on criteria described previously [26]. Immunohistochemical analysis The IMP3 antibody (L523S) was provided by Dako (Carpinteria, CA), which was a mouse monoclonal antibody (MAb) specific for the IMP3/KOC antigen. Immunohistochemical stains were performed on 5-um tissue sections from representative blocks using the purified mouse anti-IMP3 antibody and the standard avidin-biotin-complex technique as described previously [27–29]. Representative sections of endometrial serous carcinoma served as positive controls for the IMP3 antibody [29]. Negative controls were performed by replacing the primary antibody with nonimmune IgG. All slides were reviewed independently by two investigators (YW and WZ). The GSI-IX price percentage of neoplastic cells and nonneoplastic tissues that showed dark brown cytoplasmic staining was recorded. The intensity of the IHC staining was recorded as absent, weak, moderate, or strong.