First, we tested whether LCL85 Crizotinib order sensitizes mouse tumor cells to FasL induced apoptosis. Both Colon 26 and 4 T1 cells are resistant to Fas mediated apoptosis. LCL85 did not exhibit sensitization activity in Colon 26 cells to FasL induced apoptosis in our initial attempts. However, A sublethal dose of LCL85 effec tively overcame 4 T1 cells resistance to Fas mediated apoptosis. Western blotting analysis indicated that LCL85 decreased xIAP protein Inhibitors,Modulators,Libraries levels in both Colon 26 and 4 T1 cells. Toxicity of LCL85 We analyzed serum enzyme profiles to determine LCL85 liver toxicity. Analysis of serum enzyme protein levels in mice after LCL85 treatment revealed that LCL85 induces elevated alanine aminotransferase in mouse serum in a dose dependent manner, and an almost 3 fold ALT increase was detected at the highest LCL85 dose examined.
No other serum Inhibitors,Modulators,Libraries enzymes and proteins were significantly elevated by LCL85. LCL85 suppresses colon carcinoma metastatic potential in an experimental lung metastasis mouse model in vivo To determine the efficacy of LCL85 in suppression of me tastasis in vivo, we used an experimental metastasis mouse model. Colon26 cells, a highly metastatic colon carcinoma cell line, were injected i. v. to mice. Tumor bearing mice were treated with LCL85 over time. Lung metastasis was then analyzed. LCL85 significantly suppressed colon26 lung metastasis in a dose dependent manner. Although LCL85 possesses direct anti tumor cytotox icity that might contribute to the observed tumor suppression, it is possible that LCL85 might also sensitize the tumor cells to apoptosis induction by FasL of host immune cells, particularly CD8 CTLs.
We then dissected tumor bearing lungs and made single cell suspension with collagenase. Inhibitors,Modulators,Libraries Staining cells with CD8 and FasL specific mAbs revealed that CD8 T cells in tumor free mice are essentially FasL. In contrast, ap proximately 31% of tumor infiltrating CD8 T cells are FasL. CD8 cells in tumor free mice are all FasL. Therefore, LCL85 might sensitize colon carcinoma cells to host FasL CTL mediated tumor suppression. LCL85 suppresses spontaneous breast cancer metastasis in vivo To further determine the function of LCL85 in suppres sion of cancer metastasis, Inhibitors,Modulators,Libraries we used a complimentary breast cancer lung metastasis mouse model. Murine breast cancer 4 T1 cells were injected to the mammary fat pad.
Tumor bearing mice were treated with LCL85 over Inhibitors,Modulators,Libraries time and both primary tumor growth and lung metastasis were examined. LCL85 significantly suppressed the primary mammary tumor growth in vivo as measured by tumor size and tumor weight. Interestingly, the spontaneous lung metastasis was also this website significantly sup pressed by LCL85. The observation that LCL85 suppresses spontaneous breast cancer lung me tastasis is significant. However, it is possible that the decreased lung metastasis was due to the decreased primary tumor growth.