Retinoids and Retinoid Metabolism in Breast Cancer.
Abstract
My studies aim to understand vitamin A metabolism in breast tissue, specifically the role of the human 9 cis-retinol dehydrogenase (9cRDH). The 9cRDH gene was cloned from a normalized mammary library by Blaner and colleagues. We studied its expression in two normal human mammary epithelial cell strains (HMECs) and a panel of breast cancer cell lines. mRNA levels were determined by a semi-quantitative, multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) strategy. 9cRDH mRNA levels were similar among the cells tested, with the exception of the breast cancer cell lines, Hs578T and T47D, which exhibited higher expression than the normal strains, and MCF-7, which showed the lowest levels. Individual 9cRDH activity levels in normal and tumor cells, as measured by normal phase HPLC, paralleled expression levels observed by RT-PCR. In order to study the biochemical characteristics of 9cRDH and to determine optimal activity test conditions, we developed a Hep G2 cell line that expresses high levels of 9cRDH protein (LRDHSN/Hep G2). The 9cRDH cDNA was transduced into the cells using the LXSN retroviral vector containing the full length 9cRDH gene. The activity of 9cRDH tested in microsomes of LRDHSN/Hep G2 cells was time- and protein concentration-dependent and followed Michaelis-Menten kinetics.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1998
- Accession Number
- ADA357727
Entities
People
- Jisun Paik
Organizations
- University of Washington