ABSTRACT
Purpose To explore the effects of estrogen receptor antagonist on the expression of estrogen receptor subtype (ERα, ERβ), and p57kip2 protein in human endometrioid carcinoma cells named JEC. Methods The JEC cells (moderately differentiated EC cells) cultured in vitro were treated with β-Estradiol (E2) (10～6 mol/L) and two types of estrogen receptor antagonists, tamoxifen (TAM) and fulvestrant (ICI182780) (10-6 mol/L). After 24, 48, 72 h, MTT was used to detect the growth condition of JEC cells, and the light microscopy and electron microscopy were used to observe the growth condition and morphological changes of cells, Western blot was used to detect the expression of ERα, ERβ, PR-A, PR-B and P57kip2 protein in JEC cells. Results MTT results: Compared with the control group, E2 could promote the proliferation of JEC cells significantly (P＜0.05), and ICI182780 could inhibit the proliferation of JEC cells obviously (P＜0.05). Compared with the E2 group, the proliferation ability of JEC cells in E2 + ICI182780 group were lower(P＜0.05). Morphological change: Compared with the control group, the cells density of E2 group increased obviously, and the pathologic mitosis was easy to seen in some cells. The cells density decreased obviously in ICI182780 group. Compared with E2 group, the cells density of E2 + TAM group and E2 + ICI182780 group were decreased, and pathological mitotic figures were difficult to seen. Western blot results: Compared with the control group, the expression of ERβ protein increased, and the expression of p57kip2 protein decreased in E2 group (P＜0.05). The expression of ERβ protein decreased, and the expression of p57kip2 protein increased in ICI182780 group and TAM group, and the difference was statistically significant between ICI182780 group and control group (P＜0.05). Compared with the E2 group, the expression of ERβ protein decreased, and the expression of p57kip2 protein increased in E2 + ICI182780 group and E2 + TAM group, and the difference was statistically significant between E2 + ICI182780 group and E2 group (P＜0.05). ERa protein of JEC cells did not expressed in experimental group or control group. Conclusion ERa protein are not expressed in JEC cells. ICI182780 have a stronger role in antagonizing estrogen, and may induce the expression of p57kip2 protein by down-regulating the expression of ERβ protein in JEC cells, block the cell cycle progression and inhibit the growth of tumor cells. TAM has a weaker estrogen like effect on the growth of JEC cells. It is possible that combined detection of the expression of ERa and p57kip2 protein in EC has an important reference value for individualized selection of endocrine therapy for EC patients.
ABSTRACT
Objective To explore the biological effects of estrogen (17β-E2) on the proliferation and migration of human skin fibroblast (HSFB).Methods HSFBs were isolated and cultured by enzyme digestion.The fourth generation of HSFBs were adopted; (1) the proliferating effect of diverse concentrations of 17β-E2 and 17β-E2+ ICI-182780 on HSFBs was determined with MTT method at 24,48,72,96 h; (2) the influence of 17β-E2 and ICI-182780 to HSFBs cycle distribution were determined with flow cytometry; (3) the migration effect of diverse concentrations of 17β-E2 and 17β-E2+ICI-182780 on HSFBs was determined at 24,48,and 72 hours after the creation of the scratch-wound in vitro.Results (1) The proliferating speed of HSFBs in 10-10mol/L 17β-E2 group (group A)was the highest of all at 48,72,96 h,which was higher than that in ICI-182780+10-10mol/L 17β-E2 group (group B) and control group (group C) (P＜0.01) ;(2) the HSFBs during the S phase in group A was more than that in groups B and C (P＜0.01),while the HSFBs during the G0/G1 phase was less than that in groups B and C (P＜0.01); (3) the migrating effect of HSFBs in 10-8mol/L 17β-E2 group (group D) was the highest of all at 48 h,which was higher than that in ICI-182780+10-10mol/L control group (group E)and control group (group F) (P＜0.01).Conclusions The concentration of 10-10mol/L estrogen has the strongest effect of promoting proliferation and that of 10-8mol/L has the strongest chemotaxis; ICI-182780 can abate the above effect effectively.