Identification of underexpressed genes in early- and late-stage primary ovarian tumors by suppression subtraction hybridization

Vijayalakshmi Shridhar, Ami Sen, Jeremy Chien, Julie Staub, David I Smith, Steve Kovats, John Lee, Jim Lillie, David I. Smith

Research output: Contribution to journalArticle

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Abstract

To identify novel tumor suppressor genes involved in ovarian carcinogenesis, we generated four down-regulated suppression subtraction cDNA libraries from two early-stage (stage I/II) and two late-stage (stage III) primary ovarian tumors, each subtracted against cDNAs derived from normal ovarian epithelial cell brushings. Approximately 600-700 distinct clones were sequenced from each library. Comparison of down-regulated clones obtained from early- and late-stage tumors revealed genes that were unique to each library which suggested tumor-specific differences. We found 45 down-regulated genes that were common in all four libraries. We also identified several genes, the role of which in tumor development has yet to be elucidated, in addition to several under expressed genes, the potential role of which in carcinogenesis has been described previously (Bagnoli et al., Oncogene, 19: 4754-4763, 2000; Yu et al., Proc. Natl. Acad. Sci. USA, 96: 214-219, 1999; Mok et al., Oncogene, 12: 1895-1901, 1996). The differential expression of a subset of these genes was confirmed by semiquantitative reverse transcription-PCR using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as control in a panel of 15 stage I and 15 stage III tumors of mixed histological subtypes. Chromosomal sorting of library sequences revealed that several of the genes mapped to known regions of deletion in ovarian cancer. Loss of heterozygosity (LOH) analysis revealed multiple genomic regions with a high frequency of loss in both early- and late-stage tumors. To determine whether loss of expression of some of the genes corresponds to loss of an allele by LOH, we used a microsatellite marker for one of the novel genes on 8q and have shown that loss of expression of this novel gene correlates with loss of an allele by LOH. In conclusion, our analysis has identified down-regulated genes, which map to known as well as novel regions of deletions and may represent potential candidate tumor suppressor genes involved in ovarian cancer.

Original languageEnglish (US)
Pages (from-to)262-270
Number of pages9
JournalCancer Research
Volume62
Issue number1
StatePublished - Jan 1 2002

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Genes
Libraries
Loss of Heterozygosity
Neoplasms
Tumor Suppressor Genes
Oncogenes
Ovarian Neoplasms
Carcinogenesis
Clone Cells
Alleles
Gene Expression
Glyceraldehyde-3-Phosphate Dehydrogenases
Gene Library
Microsatellite Repeats
Reverse Transcription
Complementary DNA
Epithelial Cells
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Identification of underexpressed genes in early- and late-stage primary ovarian tumors by suppression subtraction hybridization. / Shridhar, Vijayalakshmi; Sen, Ami; Chien, Jeremy; Staub, Julie; Smith, David I; Kovats, Steve; Lee, John; Lillie, Jim; Smith, David I.

In: Cancer Research, Vol. 62, No. 1, 01.01.2002, p. 262-270.

Research output: Contribution to journalArticle

Shridhar, V, Sen, A, Chien, J, Staub, J, Smith, DI, Kovats, S, Lee, J, Lillie, J & Smith, DI 2002, 'Identification of underexpressed genes in early- and late-stage primary ovarian tumors by suppression subtraction hybridization', Cancer Research, vol. 62, no. 1, pp. 262-270.
Shridhar, Vijayalakshmi ; Sen, Ami ; Chien, Jeremy ; Staub, Julie ; Smith, David I ; Kovats, Steve ; Lee, John ; Lillie, Jim ; Smith, David I. / Identification of underexpressed genes in early- and late-stage primary ovarian tumors by suppression subtraction hybridization. In: Cancer Research. 2002 ; Vol. 62, No. 1. pp. 262-270.
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