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Objective To investigate the ERα (Estrogen Receptorα) Anaplastic thyroid cancer FRO cell proliferation effect and mechanism of the G protein-coupled estrogen receptor (G protein-coupled estrogen receptor, GPER) estrogen to promote. Method with 17β-estradiol (17β-Estradiol, E2), ERK1 / 2 inhibitor PD98059 (PD), Akt inhibitor LY294002 (LY), estrogen receptor inhibitor ICI182780 (ICI), built GPER small interfering RNA (GPER-siRNA) processing anaplastic thyroid carcinoma FRO cells, MTT assay of cell proliferation rate, flow cytometry cell cycle phase distribution; Western blot was used to detect FRO cells of pAkt, pERK1 / 2, GPER, ERα, of ERβ, Bcl -2, Bax protein expression. Results of different concentrations of E2 (0 mol / L, 10 -10 mol / L, and 10 -9 mol / L, 10 -8 mol / L) for anaplastic thyroid carcinoma FRO cells 24 h, cell proliferation rate were 0, (6.5 ± 1.1)%, (10.7 ± 1.9)%, (16.5 ± 2.1)% (p lt; 0.05); 10-8 mol / L the E2 deal with thyroid undifferentiated carcinoma FRO cells 0 h, 12 h, 24 h, cell proliferation rate (8.2 ± 1.9)%, (16.5 ± 2.1)% (p lt; 0.05), S / G2 / M phase cell proportion were (21.51 ± 4.32)%, (37.22 ± 4.16)%, (48.56 ± 2.31)% (p lt; 0.05); namely E2 promotes anaplastic thyroid carcinoma FRO cell proliferation and The dose-time-dependent manner. 10-8 mol / L E2 role FRO cells 0h, 12h, 24h, ERα, Bcl-2 protein expression in a time-dependent increase in Bax expression in a time-dependent reduction PD98059 and LY294002 inhibit E2 on FRO cells above role; ICI182780 inhibition of Bcl-2 protein expression, had no effect on Bax; the GPER-siRNA transfection FRO cells after 48 h, compared with the control group, GPER protein expression levels; 10 -8 mol / L of E2 treatment FRO cells at different times (0 min, 5 min, 10 min, 15 min, 30min), ERK1 / 2 and Akt phosphorylation level at 15 min and 10 min maximum; E2 FRO cells were added PD98059 (30μmol / L), LY294002 (50 μmol / L), 15 min and 10 min, compared with E2 treatment group, ERK1 / 2 and Akt phosphorylation levels decreased E2 GPER-siRNA transfection plasmid FRO cells 15 min and 10 min compared with E2 treatment group, ERK1 / 2 and Akt phosphorylation levels decreased; E2 were added PD98059, LY294002, transfected with of GPER-siRNA plasmid and ICI182780 the FRO cells 24 h compared with E2 treatment group, the rate of cell proliferation (16.5 ± 2.1)%, respectively, reduced to (11.2 ± 1.3)%, (9.6 ± 1.5)%, (7.2 ± 1.3)% and (6.9 ± 1.6)%. Conclusion E2 both by binding to the nuclear estrogen receptor ERα, the role of Bcl-2 estrogen response element, and promote the proliferation of FRO cells; also by acting GPER, activation of Akt and ERK1 / 2 pathway, thereby promoting proliferation of FRO cells.
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