In Vitro | In vitro activity: Mitomycin C, by inducing DNA interstrand crosslinks, physically blocks DNA replication, recombination, and RNA transcription. Mitomycin C potentiates TRAIL-induced apoptosis in HCT116 (p53-/-) colon cancer cells and sensitizes TRAIL-resistant colon cancer cells HT-29 to the cytokine by through JNK-independent upregulation of death receptors. In different human cancer cell lines, such OVCAR-5 (ovary), HT-29 (colon), SK-N-MC (neuroblastoma), HEP-2 (liver), COLO-205 (colon), NIH-OVCAR-3 (ovary) and A-549 (lung) cells, Mitomycin C shows cytotoxic activity.
Cell Assay: Mitomycin-C enhanced TRAIL (TNF-related apoptosis-inducing ligand)-induced apoptosis in p53 deficient colon cancer HCT116 cells. In the cell viability assay, pretreated with 5M with mitomycin C for 24h and then exposure to 25ng/ml TRAIL and 5M mitomycin C for 12h in HCT116 cells showed surprisingly decreased cell viability. In the crystal violet staining assay showed 5M mitomycin C combinated 25ng/ml TRAIL can substantially enhanced suppression effects of HCT116 cells. Pretreatment with 5M mitomycin C can enhanced TRAIL initiated processing of caspase-8, -9, -3 and cleavage of RARP (poly-ADP-ribose polymerase, substrate of caspase-3). The western blot assay showed in both HCT116 and HT29 cells, mitomycin C suppressed the expression anti apoptotic proteins Mcl-1, Bcl-2, Bcl-XL, and downregulated caspase-inhibitor c-IAP-1, XIAP, while upregulated expression of pro-apoptotic proteins Bax and Bim. |
---|