Abstract
Human kallikrein 6 (KLK6) was identified based on its transient upregulation in a primary breast tumor and its subsequent silencing in a metastatic tumor from the same patient. The molecular mechanism(s) underlying the deregulated expression of KLK6 during cancer progression are currently unknown. Here, we provide evidence that aberrant expression of KLK6 is regulated at the level of transcription by multiple cooperating mechanisms. KLK6 can be reactivated in non-expressing breast cancer cells by treatment with 5-aza-2′-deoxycytidine (5-aza-dC), a compound causing DNA demethylation. Trichostatin A (TSA), an inhibitor of histone deacetylases, resulted in moderate induction of KLK6 only in MDA-MB-231 cells. However, combined 5-aza-dC/TSA treatment resulted in synergistic activation of KLK6. We show that KLK6 inactivation is associated with hypermethylation of specific CpG dinucleotides located in the KLK6 proximal promoter and overexpression with complete demethylation. These results indicate a causal role of DNA methylation and chromatin structure in cancer-associated loss of KLK6 expression. In some breast cancer cell lines, KLK6 expression could be restored by the vitamin D3 analog EB1089. Our data indicate that transcriptional deregulation of KLK6 in cancer cells during breast cancer progression is complex and certainly not uniform in different tumors, involving epigenetic mechanisms as well as pathways regulated by nuclear receptors. This allows for the pharmacological modulation of KLK6 with potential therapeutic implications.
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