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) and Beta (ER-ß) mRNAs in Normal Ovary, Ovarian Serous Cystadenocarcinoma and Ovarian Cancer Cell Lines: Down-Regulation of ER-ß in Neoplastic Tissues
Reproductive Endocrinology Center, University of California, San Francisco, San Francisco, CA 94143-0556
Abstract
The prognosis in ovarian carcinoma, the most lethal of the gynecologic
neoplasms, is poor and has changed little in the last three decades.
Only a small number respond to antiestrogen therapy, although the
classic estrogen receptor, ER-
, has been identified in ovarian
surface epithelium, from which approximately 90% of ovarian cancers
originate. We have previously shown that ER-ß mRNA is most abundant
in human fetal ovaries, suggesting that it might play an important role
in ovarian development. Therefore, we investigated the mRNA levels of
both ERs in normal ovaries, ovarian serous cystadenocarcinomas,
granulosa cells from patients undergoing in vitro fertilization (IVF),
the ovarian surface epithelium cell line IOSE-Van, and the ovarian
cancer cell lines SKOV3, HEY and OCC1.
Northern blots of normal and neoplastic ovaries were hybridized with an
ER-ß riboprobe that spans the A/B domain. We detected two major
hybridizing bands at approximately 8 and 10 kb. An RNase protection
assay using the same probe revealed a single band of the expected size.
Hybridizing the same blot with an ER- riboprobe showed a strong
hybridizing band at approximately 6.5 kb. In ovarian cancer samples,
ER-ß mRNA level was decreased when compared to normal ovaries.
Using 25 cycles of RT-PCR followed by Southern blotting, we found equal
amounts of ER- and -ß mRNAs in normal ovaries in all age groups
from 33 to 75 years; however, in ovarian cancer tissue, the level of
ER- mRNA was similar or slightly higher, comparable to
103 to 104 copies of plasmid DNA, but ER-ß
mRNA levels were markedly decreased. Granulosa cells from IVF patients
expressed high levels of ER-ß mRNA. The OSE cell line expressed low
level of ER-, detectable after 40 cycles of RT-PCR and no ER-ß
mRNA. SKOV3, showed low level of ER- and ß mRNAs, whereas OCC1
showed low level of ER-ß and relatively high level of ER-. HEY did
not contain detectable amounts of either ER after 40 cycles of RT-PCR.
We found no evidence of differential splicing or major deletions in
almost the entire coding region of ER-ß in either normal ovaries or
tumor samples.
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