All patients had high grade and late stage cystadenocarcinomas, of the serous papillary histopathology. Patients had sub optimal surgical debulking, and all individuals died from disease progression. The cell lines selleck chem inhibitor were derived from samples collected at diagnosis and at the time of relapse, from either solid tissue or ascites. In total there were four pre chemotherapy cell lines derived from primary disease and five post chemotherapy cell lines derived from recurrent disease, OV2295, TOV2295, OV3133 and OV3133. Note that we consider TOV1369 to be a pre chemotherapy for ovarian cancer treatment, although the patient did receive chemotherapy treatment for breast cancer 18 months prior to ovarian cancer diagnosis. After 60 passages, the cell lines appeared homoge neous and no fibroblast like cells could be detected.

Although cell shape varied for each cell line, the morphology was consistent among the lines derived from the same patient samples. Figure 2 J to M Inhibitors,Modulators,Libraries shows hematoxylin and eosin staining of sections from the solid tumor tissue corresponding to cell line TOV1369, TOV2295 and TOV3133G and TOV3133D. Expression of keratin markers, TP53 and HER2 in tumor tissue and cell lines by Western blot and immunohistochemistry In order to investigate the epithelial origin of the tumors and corresponding cell lines, keratin expression was investigated by both Western blot analysis and immmu nohistochemistry. All of the keratins investigated by Western blot were present in the protein extract of each of the nine cell lines.

Expression of keratins 7, 8, 18 and 19 was also observed by immunohistochemistry using sections of the solid tumor. Keratin 20 expression was also investigated Inhibitors,Modulators,Libraries in ovarian solid tumors and from two colon cancer tissues, the latter being used as a positive control for keratin 20. No staining was observed in ovarian tissue, but Inhibitors,Modulators,Libraries positive staining was evi dent in the colon tissue. Expression of the tumor suppressor p53 Inhibitors,Modulators,Libraries was found to be present in 1369 and 2295 derived cell lines, but could not be detected in the 3133 cell lines TOV3133D, TOV3133G, OV3133 and OV3133. Western blot results for p53 were also confirmed by immunohisto chemistry, with p53 showing low expression in the TOV3133 D and TOV3133G tissues but much higher expression in TOV1369 and TOV2295. Strong HER2 expression was detected in protein extracts of all nine cell lines, and was also observed in the solid tissues by immunohistochemistry.

Mutation status of TP53, BRCA1, BRCA2, KRAS and BRAF Each of the cell lines harbored a TP53 mutation as veri fied Inhibitors,Modulators,Libraries by sequence analysis. The mutation identi fied varied with each patient sample blog post but were identical in the cell lines derived from the same patient samples. All variants are considered deleterious based on infor mation from the IARC TP53 Database.