Estrogen and Progesterone Up-Regulate Glucose Transporter Expression in ZR-75-1 Human Breast Cancer Cells

Abstract

Breast cancer incidence increases in women receiving combined estrogen and progesterone therapy. Breast tumors show increased expression of the glucose transporter GLUT1. We determined the effect of these hormones on GLUT1-4 expression and deoxyglucose transport in ZR-75-1 breast cancer cells. Immunoblotting, immunocytochemistry, flow cytometry, and RT-PCR showed that GLUT1 expression is up-regulated by progesterone and, to a greater degree, combined therapy. GLUT2 expression is unaffected by hormonal treatment. GLUT3 protein and RNA is up-regulated by progesterone and combined therapy, and GLUT4 protein expression is up-regulated by all hormonal treatments. Deoxyglucose transport studies revealed the presence of three transport components with characteristics corresponding to GLUT1/4, GLUT2, and GLUT3. 17beta-Estradiol produced a slight increase in transport at the Michaelis constant (Km) corresponding to GLUT3. Progesterone produced a small increase in transport at the Km corresponding to GLUT1/4, and combined 17beta-estradiol and progesterone produced a small increase in transport at the Km corresponding to GLUT3 and a large increase in transport at the Km corresponding to GLUT1/4. This indicates that 17beta-estradiol and progesterone differentially regulate GLUT1-4 expression and that these changes correlate to changes in glucose uptake. We postulate that combined hormone replacement therapy provides a survival advantage to developing ZR-75 breast cancer cells.

S14 protein in breast cancer cells: Direct evidence of regulation by SREBP-1c, superinduction with progestin, and effects on cell growth

“…between sex steroids and glucose in lipogenic gene regulation in breast cancer cells [41]. We found CHREBP mRNA to be readily detectable in T47D breast cancer cells, but found no major induction of S14 or FAS mRNAs in response to increasing the glucose concentration in culture media from 5.5 to 27 mM, a stimulus that is sufficient for maximal induction of lipogenesis-related genes in hepatocytes [16].…”

Differential regulation of glucose transporter expression by estrogen and progesterone in Ishikawa endometrial cancer cells

“…This theory is in line with clinical observations where estrogen therapy increased the incidence of endometrial cancer and that this increase in incidence was reduced when progesterone was added to the hormonal preparations (Smith et al 1975, Berger & Fowler 1997 and is also supported by the observation that GLUT1 is overexpressed in biopsies from cancers from the majority of estrogen and progesterone target tissues, such as breast, endometrium, cervix and ovary (Kang et al 2002, Medina & Owen 2002. Similar results are available which correlate hormonal effects on GLUT expression and glucose transport in ZR-75 breast cancer cells to breast cancer incidence (Medina et al 2003). Furthermore, overexpression of GLUT2 has not been reported in cancers originating from steroid hormone target tissues (Medina & Owen 2002), nor was an alteration in GLUT2 observed in Ishikawa cells in response to hormonal administration.…”

“…Cells were grown in DMEM/F12 media supplemented with 10% fetal bovine serum as previously published (Medina et al 2003). Briefly, depending on the experiment, cells were plated in tissue culture Petri dishes or 12-well plates (Nunc, Rochester, NY, USA), until 80% confluence, and then the medium was changed to DMEM/F12 medium containing 5% charcoal-treated serum for 24 h. Cells were divided into four groups: control, estrogen, progesterone and combined estrogen and progesterone.…”

“…3C), affected GLUT1-4 RNA expression. As a positive control of this technique for measuring hormonal regulation we used the ZR-75 breast cancer cell line (Medina et al 2003). In the ZR-75 cell line, but not in Ishikawa, treatment with progesterone, but not 17 -estradiol, regulated GLUT3 mRNA expression (Fig.…”

Upregulation of rhoA mRNA in Bronchial Smooth Muscle of Antigen-induced Airway Hyperresponsive Rats.

“…The level of rhoA mRNA was examined by RT-PCR, a simple method that is widely used for semi-quantitative measurement of mRNA levels (e.g., Niiro et al, 1997;Ma et al, 2002;Murray et al, 2002;Chen et al, 2003;Medina et al, 2003;Zhu et al, 2003). Briefly, cDNAs were prepared from the total RNA (0.5 µg) by using a Takara RNA PCR Kit (Ver.…”