摘要
Objective To study the molecular mechanism and biological significance of GPR30 activating HER2 in MCF-7 breast cancer cells with low expresses HER2.Methods Western blot was adopted to examine the phosphorylation of HER2 and the downstream signaling molecular ERK1/2 after 17-β-estradiol(E2),4-OHT(the active metabolite of tamoxifen) or G-1 (the GPR30 agonist) treatment in MCF-7 cells.After different inhibitors such as G-15 (the GPR30 antagonist),AG1478(EGFR tyrosine inhibitor),AG825 (HER2 tyrosine inhibitor),PP2 (Src family kinase inhibitor)or GM6001 (MMP inhibitor) pretreated for 2 h,the phosphorylation of HER2 and ERK1/2 were further analyzed.Finally,the altered migration and invasive capability of MCF-7 cells were detected by Transwell method.Results HER2 and ERK1/2 were activated in MCF-7 cells after E2,4-OHT or G-1 treatment and these changes could be inhibited by G-15,AG1478,AG825,PP2 or GM6001 pretreatment.The enhancement of G-1-induced migration and invasion ability in MCF-7 cells could also be inhibited by those inhibitors too.Conclusion GPR30 promotes the migration and invasion of MCF-7 cells through activating HER2-ERK1/2 signal transduction pathway.
摘要
Objective To evaluate the expression of GPR30 and Ki-67 in Non-small cell lung cancer(NSCLC) and the relationship between them.The clinicopathological features of GPR30 in NSCLC were also analyzed.The molecular mechanism that estrogen mediated the proliferation of H1299 by activating GPR30 was further studied.Methods The expression of GPR30 and Ki-67 in 80 cases of specimens of NSCLC after surgery was examined using immunohistochemistry method.After 17-β-estradiol(E2) or G-1 added,H1299 cells were counted and the cell cycle distribution was analyzed by flow cytometry.Finally,the activated ERK1/2 and the expression of cyclin D1 and p16 after G-1 treatment in H1299 cells were examined through western blotting.Results Expressions of GPR30 was more in stage Ⅲ or low differentiation tissues or adenocarcinoma (P＜0.05).A positive correlation between GPR30 and Ki-67 was further disclosed (r=0.502,P=0.000).The proliferation of H1299 cells was promoted and more cells entered S-p hase after E2 or G-1 treatment for 3 days,which could be inhibited after G-15 or U0126 pre-treatment for 2 hours.We further discovered that the activated ERK1/2 and cyclin D1 expression increased after G-1 treatment,which was blocked after G-15 or U0126 pre-treatment for 2 hours.The change of p16 was on the opposite.Conclusion A positive correlation existed between GPR30 and Ki-67.GPR30-EGFR-MAPKs signaling transduction pathway was involved in the estrogen-induced proliferation of NSCLC cells.Blocking GPR30 signaling pathway may be a promising new strategy for NSCLC treatments.
摘要
Estrogen or estrogenic substances can act via not only typical estrogen receptor α and ? but also G protein-coupled es-trogen receptor ( GPER ) . GPER is a crucial factor which can play an essential part in the non-genomic transduction pathway through which estrogen realizes its cellular functions, especially in the occurrence and treatment of estrogen related cancer. Here is a brief review concerning the current advancements on the role of GPER in estrogen related cancer.
摘要
Objective: To investigate the expression and subcellular localization of G protein-coupled receptor 30 (GPR30) in Ishikawa endometrial adenocarcinoma cells. Methods: The expressions of GPR30 mRNA and protein in Ishikawa cells were detected by RT-PCR, Western-blotting and immunofluorescence assay. The positive expression rate of GPR30 in the cell membrane and cytoplasm was determined by flow cytometry (FCM). The subcellular localization of GPR30 labeled with colloidal gold was observed under a transmission electron microscope. Results: The expressions of GPR30 mRNA and protein were both detectable and predominately localized in the cell membrane and cytoplasm of Ishikawa cells. The positive expression rate of GPR30 protein in the cytoplasm [(20.10±0.13)%] was significantly higher than that in the cell membrane [(8.54±0.17)%] (P < 0.01). The subcellular localization of GPR30 protein was mainly in the mesh-like network of cytoplasm, which may refer to rough endoplasmic reticulum. Conclusion: The expression level of GPR30 is higher in cytoplasm than in cell membrane of Ishikawa cells. GPR30 is predominantely localized in the rough endoplasmic reticulum of Ishikawa cells, which suggests an important role of GPR30 in endometrial adenocarcinoma. Copyright© 2011 by TUMOR.