| Summary: | 碩士 === 輔仁大學 === 生命科學系碩士班 === 101 === The liver cancer is the leading cause of cancer death in Taiwan. That may due to abnormal rest and sleep, drug abuse, alcoholism, smoking and overworked. The present cancer therapies still induce severely side effects. Therefore, developing a new therapy with less damage to the normal tissue and fewer side effects are attracting many researchers involved. In previous studies, some of the active ingredients in the Cruciferous can attenuate the tumor formation and induce the apoptosis signaling pathway in cancer cell. The benzyl isothiocyanate ( BITC ) is one of the extracted components that having such therapeutic potential. However, mechanism of BITC inhibit hepatoma cells is still not clear. My studies try to uncover the mechanism of BITC-induced HepG2 liver cancer cell line apoptosis.
Treated with BITC in different concentrations and time durations in HepG2 and detected by cell counting and MTT assay. The LD50 is 10 µM BITC for 24 hours and LDH assay also confirm the cytotoxicity of BITC in HepG2 cells. Annexin V-FITC positive cells proved the BITC-induced HepG2 cell apoptosis in fluorescence dye and flow cytometry analysis. Furthermore, I try to real-time detect the resistance to represent the cell survival rate by ECIS instrument. The results demonstrated that the number of cells would affect the results. After BITC treatment, undead cells would accelerate the growth rate when the cell number is too much. In WST-1 assay, pretreated with AG490 attenuated the BITC-induced HepG2 cell death. In Western blot analysis, BITC increased the PUMA protein expression. That result indicated the signaling of BITC-induced cell death. In vivo study, we induced tumor growth in nude mice and treated BITC by local injection or IP and measure the size of tumor. However, the tumor size did not significantly different after BITC treatment.
In summary, these results demonstrated that treated with 10µM BITC for 24hr maybe the optimal treatment procedure in vitro and BITC would stimulate apoptosis by JAK/STAT pathway. The size of tumor on nude mice did not significantly change after BITC treatment. That may be due to the time to start treatment with BITC.
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