It is well known that in clinic patients are under chronic stress after diagnosed with cancer prior to treatment. Thereby, in order to mimic patients in clinic as possible, sunitinib was administrated 30 minutes following NE in tests in vitro, and treatment with sunitinib was started 1 day after the implantation of pumps containing NE in tests in vivo. Tumor neovascularization or angiogenesis is closely related with proangiogenic factors such as VEGF, IL 8, IL 6, TGF and TNF released by tumor cells and immune cells. In analogy to tumors cells, lymphocytes and mac rophages in the tumor microenviroment also express B ARs triggered by NE with the following increased levels of VEGF, IL 8, and IL 6.

The NE induced up regulation of VEGF, IL 8, and IL 6 protein levels was found in a number of human cancer cell lines such as colon can cer, nasopharyngeal cancer, ovarian cancer, prostate FH535 FLT inhibitor cancer and melanoma. This effect of NE was identi fied in murine melanoma B16F1 cells and human lung adenocarcinoma A549 cells in our study. In addition, this phenomenon was also observed in murine colon cancer CT26 cells and some human cancer cells in other studies in our laboratory. However, to our knowledge, noth ing is known of the influence of NE in cancer cells treated with sunitinib in vitro. Our date indicated that, in B16F1 cells treated with sunitinib at IC50 concentration, NE also increased VEGF, IL 8, and IL 6 protein expression in culture supernatants, which could be inhibited by pro pranolol. This result offered at least a mechanism for the difference in the efficacy of sunitinib between clinical and preclinical trials.

It should be considered if sunitinib acts via some of its targets on B16 cells. Previous studies re ported that B16 cells did not express VEGFR1, VEGFR2, VEGFR3, PDGFR and PDGFRB but no more than 10% of B16 cells expressed c Kit. Whether su nitinib acts on B16 cells through the c Kit a cool way to improve target re mains to be investigated in the further study. Chronic stress has been demonstrated to promote development and progression of tumors in several human cancer cells in xenografts including prostate cancer, ovarian cancer and breast cancer, whereas no date re garding the influence of chronic stress in cancer models under sunitinib in vivo has been reported so far. This study showed that consecutive NE pumped stimulated the growth of primary tumor in a mouse melanoma model and could be blocked by propranolol.

This result provided a piece of evidence for the discrepancy in the efficacy of sunitinib between clinical and preclinical trials and was consistent with the results in the other studies in our laboratory. To further investigate stress induced angiogenesis in vivo, we analysed the immunoreactivity for VEGF and CD31, counted the MVD and measured the protein levels of VEGF, IL 8 and IL 6 in mouse serums.