Relationship between changes of the steroid receptor and synchronization in human endometrial adenocarcinoma cells in vitro.
It has been reported that the response of target cells to steroid hormone (SH) stimulation may depend on their position in the cell cycle. The DNA and RNA contents of malignant cells of the endometrium cultured in vitro were measured using flow cytometry (FCM). We also measured estrogen receptor (ER) and progesterone receptor (PR) levels of cells at different positions in the cell cycle. The G1 and S phases of the cell cycle were investigated using cells synchronized by sodium n-butyrate (G1 block), methotrexate (S block), and excess thymidine (S block). For DNA measurements, the cells were stained with propidium iodide following RNase treatment. For RNA measurements (double-stranded RNA) the cells were treated with DNase. We found that S phase synchronization by methotrexate was 136.2% of control (100%). Using the excess thymidine block and release procedure, the S phase fraction was 185.1% of control. G1 phase synchronization by sodium n-butyrate was 134% of control. The estrogen receptor level in G1 phase synchronized cells increased to 5.94 fmol/micrograms DNA in the cytosol and 12.35 fmol/micrograms DNA in the nuclear fraction. These levels represent a sevenfold total increase over that of the control estrogen receptor level. Cells in S phase showed no significant increase in estrogen receptor levels over control cells. Based on this study, the functional increase of the steroid receptor was most significant in the G1 phase.
Nishiya
I
I
Department of Obstetrics and Gynecology, Iwate Medical University, School of Medicine, Morioka, Japan.
Kagabu
T
T
Saito
S
S
Fujimoto
J
J
Okada
H
H
D000230:Adenocarcinoma / Q000032:analysis* / Q000648:ultrastructure; D002453:Cell Cycle; D002460:Cell Line; D004958:Estradiol / Q000494:pharmacology; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D011960:Receptors, Estrogen / Q000032:analysis / Q000493:pharmacokinetics; D011987:Receptors, Steroid / Q000032:analysis* / Q000187:drug effects / Q000493:pharmacokinetics; D013256:Steroids / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000032:analysis* / Q000648:ultrastructure
D004967:Estrogens; D011960:Receptors, Estrogen; D011987:Receptors, Steroid; D013256:Steroids; D004958:Estradiol