p53-Independent apoptosis by benzyl isothiocyanate in human breast cancer cells is mediated by suppression of XIAP expression.
We have shown previously that cruciferous vegetable constituent benzyl isothiocyanate (BITC) suppresses viability of cultured MCF-7 and MDA-MB-231 human breast cancer cells and retards mammary cancer development in MMTV-neu mice by causing apoptosis, but the mechanism of cell death is not fully understood. We now show that whereas p53 is dispensable for BITC-induced cell death, proapoptotic response to this promising chemopreventive agent is mediated by suppression of X-linked inhibitor of apoptosis (XIAP) protein expression. The BITC treatment increased levels of total and Ser(15)-phosphorylated p53 protein in MCF-7 cells, but the proapoptotic response to this agent was maintained even after knockdown of the p53 protein level. Exposure of MCF-7 and MDA-MB-231 cells to BITC resulted in a marked decrease in protein level of XIAP as early as 8 hours after treatment. Ectopic expression of XIAP conferred statistically significant protection against BITC-mediated cytoplasmic histone-associated apoptotic DNA fragmentation in both cell lines. Moreover, inhibition of MDA-MB-231 cell growth in vivo in female athymic mice by BITC administration correlated with a modest but statistically significant decrease in XIAP protein level in the tumor xenograft. The BITC treatment also resulted in induction as well as nuclear translocation of survivin only in the MCF-7 cells. The BITC-induced apoptosis was modestly but statistically significantly augmented by RNA interference of survivin in MCF-7 cells. In conclusion, the present study provides novel insight into the molecular circuitry of BITC-induced apoptosis to indicate suppression of XIAP expression as a critical mediator of this process.
Kim
Su-Hyeong
SH
Department of Pharmacology and Chemical Biology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pennsylvania, USA.
Singh
Shivendra V
SV
D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000230:Adenocarcinoma / Q000473:pathology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D053938:DNA Fragmentation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D055785:Gene Knockdown Techniques; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D017879:Isothiocyanates / Q000494:pharmacology*; D051379:Mice; D008819:Mice, Nude; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D011897:Random Allocation; D000077022:Survivin; D016159:Tumor Suppressor Protein p53 / Q000502:physiology; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D017879:Isothiocyanates; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D000077022:Survivin; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human; C031403:benzyl isothiocyanate