Studies have indicated the role of HSF1 (
heat-shock transcription factor 1) in repressing the transcription of some nonheat
shock genes. XAF1 (XIAP-associated factor 1) was an inhibitor of apoptosis-interacting
protein with the effect of antagonizing the cytoprotective role of XIAP. XAF1 expression was lower in
gastrointestinal cancers than in normal tissues with the mechanism unclear. Here we showed that
gastrointestinal cancer tissues expressed higher levels of HSF1 than matched normal tissues. The expression of XAF1 and HSF1 was negatively correlated in
gastrointestinal cancer cell lines. Stress stimuli, including heat, hypo-osmolarity, and H2O2, significantly suppressed the expression of XAF1, whereas the alteration of HSF1 expression negatively correlated with XAF1 expression. We cloned varying lengths of the 5'-flanking region of the XAF1 gene into
luciferase reporter vectors, and we evaluated their promoter activities. A transcription silencer was found between the -592- and -1414-nucleotide region that was rich in nGAAn/nT-TCn elements (where n indicates G, A, T, or C). A high affinity and functional HSF1-binding
element within the -862/-821-
nucleotide region was determined by electrophoretic mobility shift assay and
chromatin immunoprecipitation assay. Inactivation of this "heat-shock
element" by either site-directed mutation or an HSF1 inhibitor,
pifithrin-alpha, restored the promoter activity of the silencer structure. Moreover, pretreatment with
antioxidants suppressed HSF1 binding activity and increased the transcriptional activity and expression of XAF1. These findings suggested that endogenous stress pressure in
cancer cells sustained the high level expression of HSF1 and subsequently suppressed XAF1 expression, implicating the synergized effect of two
anti-apoptotic protein families, HSP and inhibitors of apoptosis, in cytoprotection under stress circumstances.