Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1997-03-17
1999-11-02
Ketter, James
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 243, C12Q 168, C07H 2104
Patent
active
059767995
ABSTRACT:
Increased expression of the p16 gene occurs early in the development of ovarian carcinomas. This invention detects change ovarian epithelium by measuring increases in p16 gene expression by a quantitative method that compares the levels of p16 mRNA and a control mRNA (.beta.-tubulin) in a subject to be tested against the levels of substrates in normal subjects. A biological sample such as peritoneal fluid containing mRNA derived from a subject's ovarian epithelium is taken from the subject to be tested. The mRNA is isolated from the sample, and complementary cDNA is prepared from the isolated mRNA. Using primers to p16 target sequences and to .beta.-tubulin control sequences, the cDNA is amplified. The resultant amplification products are quantitated as to p16 and .beta.-tubulin gene sequences. The level p16 gene expression is assessed relative to expression levels in normal subjects. An increased level of p16 gene expression in a subject determined by this method is an indication of change in the subject's ovarian epithelium indicative of presence of a carcinoma.
REFERENCES:
Rodabaugh et al. Detailed deletion mapping of chromosome 9p and p16 gene alterations in human borderline and invasive epithelial ovarian tumors. Oncogene vol. 11:1249-1254, Nov. 2, 1995.
Ragione et al. 5'-deoxy-5'-methyladenosine phosphorylase and p 16(ink4) deficiency in multiple tumor cell lines. Oncogene vol. 10:827-833, Mar. 16, 1995.
Okamoto et al. p16(ink4) mutations and altered expression in human tumors and cell lines. Cold Spring Harbor Symposia on Quantitative Biology. vol. 59:49-57, 1994.
Schultz et al. Characterization of chromosome 9 in human ovarian neoplasia identifies frequent genetic imbalance on 9q and rare alterations involving 9p, including CDKN2. Cancer Res. vol. 55:2150-2157, May 15, 1995.
O'Brien Timothy J.
Shiqemasa Kazushi
Ketter James
McGregor Martin L.
Sandals William
The Board of Trustees of the University of Arkansas
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