Epigenetic silencing of the tumor suppressor cystatin M occurs during breast cancer progression

Lingbao Ai, Wan Ju Kim, Tae You Kim, C. Robert Fields, Nicole A. Massoll, Keith D Robertson, Kevin D. Brown

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Cystatin M is a secreted inhibitor of lysosomal cysteine proteases. Several lines of evidence indicate that cystatin M is a tumor suppressor important in breast malignancy; however, the mechanism(s) that leads to inactivation of cystatin M during cancer progression is unknown. Inspection of the human cystatin M locus uncovered a large and dense CpG island within the 5′ region of this gene (termed CST6). Analysis of cultured human breast tumor lines indicated that cystatin M expression is either undetectable or in low abundance in several lines; however, enhanced gene expression was measured in cells cultured on the DNA demethylating agent 5-aza-2′-deoxyeytidine (5-aza-dC). Increased cystatin M expression does not correlate with a cytotoxic response to 5-aza-dC; rather, various molecular approaches indicated that the CST6 gene was aberrantly methylated in these tumor lines as well as in primary breast tumors. Moreover, 60% (12 of 20) of primary tumors analyzed displayed CST6 hypermethylation, indicating that this aberrant characteristic is common in breast malignancies. Finally, preinvasive and invasive breast tumor cells were microdissected from nine archival breast cancer specimens. Of the five tumors displaying CST6 gene methylation, four tumors displayed methylation in both ductal carcinoma in situ and invasive breast carcinoma lesions and reduced expression of cystatin M in these tumors was confirmed by immunohistochemistry. In summary, this study establishes that the tumor suppressor cystatin M is a novel target for epigenetic silencing during mammary tumorigenesis and that this aberrant event can occur before development of invasive breast cancer.

Original languageEnglish (US)
Pages (from-to)7899-7909
Number of pages11
JournalCancer Research
Volume66
Issue number16
DOIs
StatePublished - Aug 15 2006
Externally publishedYes

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Cystatin M
Epigenomics
Breast Neoplasms
Neoplasms
Breast
Methylation
Minor Lymphocyte Stimulatory Loci
Genes
Cysteine Proteinase Inhibitors
CpG Islands
Carcinoma, Intraductal, Noninfiltrating

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Epigenetic silencing of the tumor suppressor cystatin M occurs during breast cancer progression. / Ai, Lingbao; Kim, Wan Ju; Kim, Tae You; Fields, C. Robert; Massoll, Nicole A.; Robertson, Keith D; Brown, Kevin D.

In: Cancer Research, Vol. 66, No. 16, 15.08.2006, p. 7899-7909.

Research output: Contribution to journalArticle

Ai, Lingbao ; Kim, Wan Ju ; Kim, Tae You ; Fields, C. Robert ; Massoll, Nicole A. ; Robertson, Keith D ; Brown, Kevin D. / Epigenetic silencing of the tumor suppressor cystatin M occurs during breast cancer progression. In: Cancer Research. 2006 ; Vol. 66, No. 16. pp. 7899-7909.
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abstract = "Cystatin M is a secreted inhibitor of lysosomal cysteine proteases. Several lines of evidence indicate that cystatin M is a tumor suppressor important in breast malignancy; however, the mechanism(s) that leads to inactivation of cystatin M during cancer progression is unknown. Inspection of the human cystatin M locus uncovered a large and dense CpG island within the 5′ region of this gene (termed CST6). Analysis of cultured human breast tumor lines indicated that cystatin M expression is either undetectable or in low abundance in several lines; however, enhanced gene expression was measured in cells cultured on the DNA demethylating agent 5-aza-2′-deoxyeytidine (5-aza-dC). Increased cystatin M expression does not correlate with a cytotoxic response to 5-aza-dC; rather, various molecular approaches indicated that the CST6 gene was aberrantly methylated in these tumor lines as well as in primary breast tumors. Moreover, 60{\%} (12 of 20) of primary tumors analyzed displayed CST6 hypermethylation, indicating that this aberrant characteristic is common in breast malignancies. Finally, preinvasive and invasive breast tumor cells were microdissected from nine archival breast cancer specimens. Of the five tumors displaying CST6 gene methylation, four tumors displayed methylation in both ductal carcinoma in situ and invasive breast carcinoma lesions and reduced expression of cystatin M in these tumors was confirmed by immunohistochemistry. In summary, this study establishes that the tumor suppressor cystatin M is a novel target for epigenetic silencing during mammary tumorigenesis and that this aberrant event can occur before development of invasive breast cancer.",
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