Yu Tenghua,Luo Haojun,Yan Yuzhao,Wu Chengyi,Liu Manran,Tu Gang.Effects of estrogen on GPR30-mediated proliferation in triple-negative breast cancer cell line MDA-MB-468[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(14):1467-1471.
Yu Tenghua,Luo Haojun,Yan Yuzhao,Wu Chengyi,Liu Manran,Tu Gang.Effects of estrogen on GPR30-mediated proliferation in triple-negative breast cancer cell line MDA-MB-468[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(14):1467-1471.DOI:
Effects of estrogen on GPR30-mediated proliferation in triple-negative breast cancer cell line MDA-MB-468
Objective To explore the effects of estrogen on the proliferation of triple-negative breast cancer cell line MDA-MB-468 mediated by G protein-coupled estrogen receptor GPR30. Methods Fluorescence immunoassay and W
estern blotting were performed to examine the localization and expression of GPR30 in MDA-MB-468 cells. Cell cycle and cell proliferation were tested by flow cytometry and CCK-8 assay. The expression of phospho-extracellular regulate kinase (p-ERK) and Cyclin D1 was detected by Western blotting. Results GPR30 was detected with high expression level in the MDA-MB-468 cells and located mostly in the cytoplasm. After treatment with 17-β-estradiol (E
2
)
GPR30 specific agonist (G1) and tamoxifen (TAM)
the progression of cell cycle and cell proliferation was increased remarkably. The DNA contents in DNA synthesis (S) phase were 2.81±0.11
2.82±0.21
and 2.70±0.20 times higher and the relative cell numbers were 1.83±0.18
1.94±0.12
and 1.92±0.16 times higher than those of the control group
respectively. The above effects induced by E
2
G1 and TAM could be blocked by GPR30 specific antagonist G15 (
P
0.05). The relative protein level of p-ERK in the E
2
G1 and TAM treatment groups was 2.59±0.21
2.43±0.25
and 2.26±0.34 times higher and that of cyclin D1 was 1.67±0.06
1.51±0.08
1.90±0.07 times higher than those of the control group
respectively (
P
0.05). Interestingly
the changes induced by E
2
G1 and TAM were inhibited by G15 (
P
0.05). Conclusion Estrogen triggers downstream ERK signaling by activating GPR30 to up-regulate the expression of p-ERK and cyclin D1
which accelerates MDA-MB-468 cell cycle progression