[Epigenetic inactivation of the WNT antagonist SFRP1 in breast cancer]

The WNT signalling pathway plays a central role during embryonic development of multi-cellular organisms, especially for the temporal and spatial specification of organs (e.g. WNT4 in kidney development), a process called pattern formation. Interestingly, genes of the WNT pathway are deregulated in a variety of solid tumours, being considerably up- or down-regulated compared to their expression in the corresponding normal tissue. Some members like WNT1 have demonstrated oncogenic properties in animal models. The SFRP1 gene on chromosome 8 p12 is a negative regulator of the WNT pathway. SFRP1 protein is supposed to bind WNT1 molecules thereby inhibiting the activation of frizzled receptors and the WNT pathway. Characterising an SFRP1-specific antibody we could show that loss of SFRP1 is an extremely common event in breast cancer, i.e. SFRP1 was considerably down-regulated in 73% (n = 1967) of analysed invasive breast cancers. SFRP1 loss is associated with unfavourable prognosis in early breast cancer (pT1 tumours). To analyse the cause of SFRP1 inactivation in breast cancer we performed a parallel expression and promoter methylation analysis in human breast cancer and tumour cell lines. RT-PCR techniques and methylation-specific PCR (MSP) were applied. All tumorigenic cell lines analysed exhibited complete promoter methylation and did not express detectable amounts of SFRP1 mRNA. SFRP1 expression could be restored in these cell lines after treatment with 5-Aza-2'-deoxycytidine, a demethylating agent. Human primary breast cancer was methylated in nearly 75% of cases. Our results indicate that epigenetic inactivation by methylation is the predominant mechanism of SFRP1 gene silencing in breast cancer.