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17ß-Hydroxysteroid Dehydrogenase Gene Expression in Human Breast Cancer Cells: Regulation of Expression by a Progestin¹

Biocenter and Department of Clinical Chemistry, University of Oulu, ³ SF-90220 Oulu, Finland

The expression of the 17ß-hydroxysteroid dehydrogenase (17-HSD) gene in a series of human breast cancer cell lines was studied by Northern blot hydridization with a cDNA probe and by a time-resolved immunofluorometric assay using polyclonal antibodies against the enzyme protein. The 17-HSD enzyme protein concentration was measured in the 800 x g cell extract. A high concentration was measured in the BT-20 cell line, corresponding to one-fourth of the average concentration in placental tissue. Western blot analysis indicated that the antigen corresponded to a single M_r 35,000 band. In 2 other cell lines (MDA-MB-361 and T-47D), the 17-HSD protein concentration was much lower, but still measurable, whereas in the remaining 5 cell lines (HBL-100, MCF-7, MDA-MB-231, MDA-MB-468, and ZR-75-1) it was below the detection limit of the assay. Treatment of the cells for 5 days with the synthetic progestin, ORG2058, resulted in an increase of the 17-HSD protein concentration only in the T-47D cell line. By Northern blot analysis, a low level of 2.3-kilobase mRNA transcripts was detected in all 8 cell lines. In addition, a 1.3-kilobase 17-HSD mRNA was present in the samples from the 3 cell lines containing measurable amounts of 17-HSD protein in the cell extract, and the band intensities were proportional to the amount of protein measured with the immunofluorometric assay. Only in the T-47D cell line did progestin treatment correspond to an increased amount of the 17-HSD 1.3-kilobase mRNA. These results suggest that the 1.3-kilobase mRNA for 17-HSD is the form most closely associated with protein expression and is also the only form responding to the progestin induction of the 17-HSD gene.

¹ This work was supported by the Research Council for Medicine of the Academy of Finland and by the Foundation of the Finnish Cancer Institute.

² To whom requests for reprints should be addressed, at: Biocenter and Department of Clinical Chemistry, University of Oulu, SF-90220 Oulu, Finland.

³ The Department of Clinical Chemistry is a WHO Collaborating Centre for Research in Human Reproduction supported by the Ministries of Education, of Health and Social Affairs, and of Foreign Affairs, Finland.

Received 7/10/91. Accepted 10/28/91.

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