Inactivation of the cystatin E/M tumor suppressor gene in cervical cancer

Mysore S. Veena, Grant Lee, Daniel Keppler, Marc Mendonca, J. Leslie Redpath, Eric J. Stanbridge, Sharon P. Wilczynski, Eri S. Srivatsan

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We have previously localized a cervical cancer tumor suppressor gene to a 300 kb interval of 11q13. Analysis of candidate genes revealed loss of expression of cystatin E/M, a lysosomal cysteine protease inhibitor, in 6 cervical cancer cell lines and 9 of 11 primary cervical tumors. Examination of the three exons in four cervical cancer cell lines, 19 primary tumors, and 21 normal controls revealed homozygous deletion of exon 1 sequences in one tumor. Point mutations were observed in six other tumors. Two tumors contained mutations at the consensus binding sites for cathepsin L, a lysosomal protease overexpressed in cervical cancer. Introduction of these two point mutations using site directed mutagenesis resulted in reduced binding of mutated cystatin E/M to cathepsin L. Although mutations were not observed in any cell lines, four cell lines and 12 of 18 tumors contained promoter hypermethylation. Reexpression of cystatin E/M was observed after 5′aza 2-deoxycytidiene and/or Trichostatin A treatment of cervical cancer cell lines, HeLa and SiHa, confirming promoter hypermethylation. Ectopic expression of cystatin E/M in these two cell lines resulted in growth suppression. There was also suppression of soft agar colony formation by HeLa cells expressing the cystatin E/M gene. Reexpression of cystatin E/M resulted in decreased intracellular and extracellular expression of cathepsin L. Overexpression of cathepsin L resulted in increased cell growth which was inhibited by the reintroduction of cystatin E/M. We conclude, therefore, that cystatin E/M is a cervical cancer suppressor gene and that the gene is inactivated by somatic mutations and promoter hypermethylation.

Original languageEnglish
Pages (from-to)740-754
Number of pages15
JournalGenes Chromosomes and Cancer
Volume47
Issue number9
DOIs
StatePublished - Sep 2008

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Cystatin M
Tumor Suppressor Genes
Uterine Cervical Neoplasms
Cathepsin L
Cell Line
Neoplasms
Point Mutation
Mutation
Exons
trichostatin A

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

Cite this

Veena, M. S., Lee, G., Keppler, D., Mendonca, M., Redpath, J. L., Stanbridge, E. J., ... Srivatsan, E. S. (2008). Inactivation of the cystatin E/M tumor suppressor gene in cervical cancer. Genes Chromosomes and Cancer, 47(9), 740-754. https://doi.org/10.1002/gcc.20576

Inactivation of the cystatin E/M tumor suppressor gene in cervical cancer. / Veena, Mysore S.; Lee, Grant; Keppler, Daniel; Mendonca, Marc; Redpath, J. Leslie; Stanbridge, Eric J.; Wilczynski, Sharon P.; Srivatsan, Eri S.

In: Genes Chromosomes and Cancer, Vol. 47, No. 9, 09.2008, p. 740-754.

Research output: Contribution to journalArticle

Veena, MS, Lee, G, Keppler, D, Mendonca, M, Redpath, JL, Stanbridge, EJ, Wilczynski, SP & Srivatsan, ES 2008, 'Inactivation of the cystatin E/M tumor suppressor gene in cervical cancer', Genes Chromosomes and Cancer, vol. 47, no. 9, pp. 740-754. https://doi.org/10.1002/gcc.20576
Veena, Mysore S. ; Lee, Grant ; Keppler, Daniel ; Mendonca, Marc ; Redpath, J. Leslie ; Stanbridge, Eric J. ; Wilczynski, Sharon P. ; Srivatsan, Eri S. / Inactivation of the cystatin E/M tumor suppressor gene in cervical cancer. In: Genes Chromosomes and Cancer. 2008 ; Vol. 47, No. 9. pp. 740-754.
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