vehicle treated tumors was evaluated by IHC staining for the active form of caspase 3, cleaved caspase 3, having an antibody that recognizes the subunit in the cytoplasm of apoptotic cells. Just rare positive cells were determined in tissue sections from tumors treated with rapamycin or car, and no significant difference was noted between the two groups. This PF299804 molecular weight finding is in line with previous studies that rapamycin and its analogs may sensitize tumefaction cells in culture to cisplatin induced apoptosis, but have minimal effects on apoptosis when used alone. Ramifications of paclitaxel and cisplatin on tumor cell proliferation and apoptosis could not be examined because residual tumor was identified in just one of six treated animals. Immunoblotting and IHC staining were used to research residual APC?/PTEN? tumors remaining after four weeks of treatment with rapamycin. Only small levels of tumor tissue remained after-treatment, limiting how many studies that could be performed. We discovered that pS6 levels were lower, and pAKT Gene expression levels slightly increased, in rapamycintreated tumors compared to those receiving vehicle. IHC staining of residual tumefaction tissue confirmed significant reduction of pS6 within the rapamycin treated cancers when compared with controls. Tumor imaging The capability to non-invasively and quantitatively image localized and metastatic OEAs in live animals would allow precise and repeated measurements of tumor burden, increasing statistical power and reducing the number of animals needed to try each therapeutic regimen. To show the feasibility of this method, we further engineered our OEA model to add a luciferase reporter allele which can be activated by AdCre. Mice with a Cre activatable kind of firefly luciferase allele present at the ubiquitously expressed AG-1478 ic50 Rosa26 locus were crossed with Apcflox/flox,Ptenflox/flox mice to generate Apcflox/flox,Ptenflox/flox,ROSA26L S L Luc/ mice. We performed ovarian bursal shot of AdCre in Apcflox/flox,Ptenflox/flox,Rosa26L S D Luc/ mice and bioluminescence imaging was used to monitor tumor response to rapamycin therapy over an one-month course of treatment. Two tumor bearing mice were treated with rapamycin and two were treated with vehicle. BLI was carried out before initiation of treatment 6 months after ovarian bursal injection of AdCre, and weekly for one month thereafter. Equally car treated animals showed a substantial increase in tumor bioluminescence over the treatment period, while bioluminescence in the rapamycin treated mice reduced in the other mouse and increased only minimally in one mouse. Comparison of tumefaction size and BLI signal at study end-point is shown in Figure 5G. MEK/ERK signaling is up-regulated in reaction to AKT inhibition in murine APC?/PTEN? and human ovarian carcinoma cell lines Recent findings suggest a connection between mTOR inhibition and ERK activation, perhaps reflecting interruption of an S6K1 dependent negative feedback loop.