Abstract
Cystatin M is a potent endogenous inhibitor of lysosomal cysteine proteases. In breast carcinoma, cystatin M expression is frequently downregulated. It has been shown that cystatin M expression suppressed growth and migration of breast cancer cells. We examined the methylation status of the CpG island promoter of cystatin M in four breast cancer cell lines (MDAMB231, ZR75-1, MCF7 and T47D), in 40 primary breast carcinoma and in corresponding normal tissue probes by combined bisulphite restriction analysis. To investigate the effects of cystatin M expression on the growth of breast carcinoma, cystatin M was transfected in T47D. The cystatin M promoter was highly methylated in all four-breast cancer cell lines. Primary breast tumours were significantly more frequently methylated compared to normal tissue samples (60 vs 25%; P=0.006 Fisher's exact test). Treatment of breast cancer cells with 5-aza-2′-deoxycytidine (5-Aza-CdR), reactivated the transcription of cystatin M. Transfection of breast carcinoma cells with cystatin M caused a 30% decrease in colony formation compared to control transfection (P=0.002). Our results show that cystatin M is frequently epigenetically inactivated during breast carcinogenesis and cystatin M expression suppresses the growth of breast carcinoma. These data suggest that cystatin M may encode a novel epigenetically inactivated candidate tumour suppressor gene.
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Abbreviations
- COBRA:
-
combined bisulphite restriction analysis
- 5-Aza-CdR:
-
5-aza-2′-deoxycytidine
- RASSF1A :
-
Ras association domain family 1A
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Acknowledgements
This study was supported by Grants BMBF FKZ01ZZ0104 and DFG DA552-1 to Reinhard Dammann and WR (FKZ13/13) to Undraga Schagdarsurengin and NIH Grant CA88873 to Gerd P Pfeifer.
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Schagdarsurengin, U., Pfeifer, G. & Dammann, R. Frequent epigenetic inactivation of cystatin M in breast carcinoma. Oncogene 26, 3089–3094 (2007). https://doi.org/10.1038/sj.onc.1210107
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DOI: https://doi.org/10.1038/sj.onc.1210107
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