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PMID: 10733592 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

17beta-estradiol inhibits apoptosis in MCF-7 cells, inducing bcl-2 expression via two estrogen-responsive elements present in the coding sequence.

Molecular and cellular biology ·Vol. 20 ·No. 8 ·2000-04-00 ·Pages 2890-901

Perillo B, Sasso A, Abbondanza C, Palumbo G

Abstract

We have found that 17beta-estradiol induces bcl-2 transcription in human breast cancer MCF-7 cells. To identify cis-acting elements involved in this regulation, we have analyzed hormone responsiveness of transiently transfected reporter constructs containing the bcl-2 major promoter (P(1)). Hormone inducibility was observed only when either of two sequences, located within the bcl-2 coding region and showing one and two mutations with respect to the consensus estrogen-responsive element, were inserted downstream from the P(1) promoter. Both sequences behaved as enhancers exclusively in cells expressing the estrogen receptor and were able to bind this receptor in in vitro assays. Transfections into MCF-7 cells of plasmids carrying a bcl-2 cDNA fragment which included these two elements revealed that their simultaneous presence resulted in an additive effect on reporter gene activity, whose size resembled the increase of endogenous bcl-2 mRNA level observed in untransfected cells after hormone treatment. Moreover, the identified elements were able to mediate up-regulation of bcl-2 expression by 17beta-estradiol, since exogenous bcl-2 mRNA was induced by hormone challenge of MCF-7 cells transiently transfected with a vector containing the bcl-2 coding sequence cloned under the control of a non-estrogen-responsive promoter. Finally, we show that hormone prevention of apoptosis, induced by incubating MCF-7 cells with hydrogen peroxide, was strictly related to bcl-2 up-regulation. Our results indicate that the bcl-2 major promoter does not contain cis-acting elements directly involved in transcriptional control by 17beta-estradiol and that hormone treatment inhibits programmed cell death in MCF-7 cells, inducing bcl-2 expression via two estrogen-responsive elements located within its coding region.

MeSH Terms
Apoptosis/drug effects,genetics Binding Sites/genetics Breast Neoplasms/genetics,pathology Estradiol/pharmacology Female Gene Expression Regulation, Neoplastic/drug effects Genes, bcl-2 Humans Tumor Cells, Cultured
Chemicals
Estradiol
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Perillo B
Centro di Endocrinologia ed Oncologia Sperimentale del C.N.R., Dipartimento di Biologia e Patologia Cellulare e Molecolare "L. Califano", Facoltà di Medicina e Chirurgia, Università "Federico II", 80131 Naples, Italy. perillo@unina.it
Sasso A
Abbondanza C
Palumbo G
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.20.8.2890-2901.2000
Email
perillo@unina.it
Published
2000-04-00
Pages
2890-901
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC85519
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.20.8.2890-2901.2000 PMC Full Text View journal details
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