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Messenger RNA differential display reverse‐transcriptase‐polymerase‐chain‐reaction analysis of a progestogen‐suppressive gene in a human endometrial‐cancer cell line
Author(s) -
Sakata Masahiro,
Kurachi Hirohisa,
Morishige Kenichirou,
Ogura Kazuhiro,
Yamaguchi Masaaki,
Nishio Yukihiro,
Ikegami Hiromasa,
Miyake Akira,
Murata Yuji
Publication year - 1998
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19980925)78:1<125::aid-ijc20>3.0.co;2-9
Subject(s) - differential display , biology , northern blot , microbiology and biotechnology , reverse transcription polymerase chain reaction , progestogen , endometrium , reverse transcriptase , messenger rna , gene expression , endometrial cancer , endocrinology , gene , estrogen , cancer , polymerase chain reaction , genetics
Progestogen suppresses the progression of endometrial cancer and has an important effect on the secretory change of human endometrium. We characterized the progestogen‐induced alterations of gene expression in a human endometrial‐cancer cell line using a mRNA differential‐display reverse‐transcriptase‐polymerase‐chain‐reaction (DDRT‐PCR) method. After 5‐day incubation of Ishikawa endometrial‐cancer cells, with or without 100 nM medroxyprogesterone acetate (MPA), total RNA was isolated from confluent cells. We identified 8 candidate genes by mRNA differential display by screening up to approximately 3,000 mRNA species. Among these, 2 genes named T21A and T21B showed a decrease in mRNA by MPA treatment when analyzed by Northern blot. Nucleotide sequence showed that clone T21A was part of human mitochondrial short‐chain enoyl‐CoA hydratase cDNA. The other clone, T21B, showed no homology with any known nucleotide sequences. Northern‐blot analysis using T21A and T21B clones as probes showed a decrease in mRNA in human endometrium from the luteal stage, with high serum estradiol and progesterone levels, as compared with that from the early follicular stage, with low serum estradiol and progesterone levels, and that from the pre‐ovulatory stage with high serum estradiol and low progesterone levels. These findings suggest that mRNA DDRT‐PCR could be used to identify the candidate genes regulated by progestogen in human endometrial cancer and in normal human endometrium. Int. J. Cancer 78:125–129, 1998.© 1998 Wiley‐Liss, Inc.