FIG. 3.
Effect of biochanin A (BCA) and other phytoestrogens/steroids on CCFSTTG1 and U87MG cells viability and p75NTR expression levels in a prevention regimen. MTT relative cell viability of CCFSTTG1 (A) and U87MG (C) cells treated with 0, 10, 20, 40, 60, 80, or 100 μM BCA or genistein (GEN), or 0, 10, 20, 40, 60, 80, or 100 nM dexamethsone (DEX) or estradiol (E2), for 24 h followed by treatment with ibuprofen (2 mM) + 0, 10, 20, 40, 60, 80, or 100 μM BCA or genistein, or 0, 10, 20, 40, 60, 80, or 100 nM DEX for an additional 24 h. Data are presented as mean cell survival ± standard error of the mean (SEM), normalized to ibuprofen-treated cells only, with * and ** indicating p < 0.05 and 0.01, respectively, as compared to ibuprofen only-treated cells. Representative Western blots of p75NTR levels in CCFSTTG1 (B) and U87MG (D) cells treated similarly as above, cells were lysed and similar amounts of protein (40 μg) were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblots were reprobed for β-actin to ensure for equal loading; A875 melanoma cell lysates were used as a positive control for p75NTR. Respective quantifications (from two independent experiments) of p75NTR expression are shown below as mean ± SEM for every cell type and normalized to levels in ibuprofen-treated cells.