Cell cycle phase dependence of estrogen and epidermal growth factor (EGF) receptor expression in MCF-7 cells: implications in antiestrogen and EGF cell responsiveness.
In this report the effect of epidermal growth factor (EGF) and the antiestrogen hydroxytamoxifen (OH-TAM) on the cell cycle of the breast cancer cell line MCF-7 was investigated as a function of the presence of their respective receptors. For this study synchronized cells were obtained by cell incubation in the presence of 2 mM thymidine for 24 h at 37 C. The treatment led to a partial synchronization, since at the end of thymidine treatment, 80% of cells were accumulated in the G1 phase. The removal of thymidine allowed the cells to progress through the cell cycle, since between 6-9 h after the arrest of the treatment, about 50% of cells were found in the S phase. By 9-12 h, most of the cells entered the G2 phase, and by 24 h, the cells returned to the G1 phase. When MCF-7 cells were incubated in the presence of OH-TAM for various periods of time before thymidine exposure, the progression of the cells through the cell cycle was dramatically inhibited. Also, a short term antiestrogen treatment (2 h) before or immediately after the addition of thymidine led to an accumulation of MCF-7 cells in the G1 phase. However, when the cells were treated for 2 h with OH-TAM 22 h after thymidine addition or shortly after its removal from the cell culture, no effect of the antiestrogen on the cell cycle could be observed. In parallel, the effect of thymidine on the level of estrogen receptor was studied. Although low affinity estrogen-binding sites were maintained, high affinity ER were found to be dramatically reduced during the thymidine treatment. The comparison between the effect of OH-TAM on the cell cycle and the expression of ER revealed that the antiestrogen OH-TAM was effective only in the presence of ER. EGF was found to have no effect on the cell cycle of thymidine-synchronized cells, although it did partially reverse the G1 phase block induced by OH-TAM when added simultaneously to cell culture 24 h before thymidine exposure. The parallel analysis of EGF receptor level demonstrated that thymidine treatment also reduced EGF receptors that were found to reappear after the synchronization, during the S and G2 phases of the cell cycle.(ABSTRACT TRUNCATED AT 400 WORDS)
Laboratoire Cancérologie Expérimentale, CNRS SDI 6194, Faculté Médecine Secteur Nord, Marseille, France.
X F
XF
Dong
Y
Y
Berthois
E
E
Colomb
P M
PM
Martin
D001665:Binding Sites; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002453:Cell Cycle / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D066246:ErbB Receptors / Q000378:metabolism*; D004965:Estrogen Antagonists / Q000494:pharmacology*; D004967:Estrogens / Q000378:metabolism; D011960:Receptors, Estrogen / Q000378:metabolism*; D013936:Thymidine / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D004965:Estrogen Antagonists; D004967:Estrogens; D011960:Receptors, Estrogen; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D013936:Thymidine