Trichosanthin-induced apoptosis in melanoma B-16 cell line / 中国病理生理杂志

ABSTRACT

AIM: To investigate the antitumor effect and mechanism of trichosanthin (TCS) on melanoma B-16 cells. METHODS: (1) The injury of B-16 cells by trichosanthin was observed with SCGE and hoechst33258 staining method. (2) LCSM and specificity fluorescent probe Fluo-3/AM, H2DCF-DA, DAF-FM diacetate were applied to analyze the dynamic changes of Ca2+, ROS and NO in single cell cultured with TCS. Simultaneously, the relationship between ROS, NO and increase of Ca2+ was also revealed. RESULTS: (1)When treated with TCS (50 mg/L) for 3 h and 6 h, neither cytotoxicity assay nor SCGE showed the differences compared with control group. After 12 h incubation, specificity phenomena of DNA injury-comet tail appeared in SCGE and chromatin condensation even apoptotic body formation were seen by Hoechst33258 staining. (2) TCS (50 mg/L) evoked rapid enhancement of the production of Ca2+, ROS and NO in the cell and the differences between TCS and control group had statistical significance (P