title: [GPER silence inhibits the stimulation of growth and inhibition of apoptosis induced by tamoxifen in breast cancer-associated fibroblasts].
abstract: OBJECTIVE
  To construct a lentiviral vector (Lenti-GPER-shRNA) targeting G-protein coupled estrogen receptor (GPER) and explore the role of GPER in the effect of tamoxifen on cell proliferation and apoptosis in breast cancer associated fibroblasts (BCAFs).
  
  
  METHODS
  The target sequence of GPER gene and negative control were cloned into lentiviral vectors. The recombinant lentivirus and control were extracted after HEK293T cells were transfected with the recombinant vector and helper vectors. After infection of BCAFs with the GPER lentiviral vector under the best interfering condition, GPER expression was detected by real-time quantitative PCR and Western blotting. BCAFs were divided into negative control group, GPER-RNAi group, negative control combined with tamoxifen (10(-8) mmol/L) group and GPER-RNAi combined with tamoxifen (10(-8) mmol/L) group. CCK-8 assay was used to detect the proliferation and annexin V-fluorescein isothiocyanate/propidium iodide (annexin V-FITC/PI) combined with flow cytometry was used to detect the apoptosis of BCAFs after the treatment of tamoxifen.
  
  
  RESULTS
  Lenti-GPER-shRNA significantly interfered the expression of GPER in BCAFs. Tamoxifen promoted the growth of BCAFs, which could be attenuated by knockdown of GPER. Moreover, the apoptosis of BCAFs was reduced by tamoxifen, which was also reversed by knockdown of GPER.
  
  
  CONCLUSIONS
  Lenti-GPER-shRNA could effectively silence the GPER expression in BCAFs. The ability of tamoxifen to accelerate cell proliferation and decrease cell apoptosis could be weakened by knockdown of GPER.
authors.affiliation: Department of Endocrine and Breast Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Yuzhao
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Endocrine and Breast Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Tenghua
authors.initials: T
authors.lastname: Yu
authors.affiliation: Department of Endocrine and Breast Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Tu
authors.affiliation: Ministry of Education Key Laboratory of Clinical Diagnosis, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Manran
authors.initials: M
authors.lastname: Liu
authors.affiliation: Department of Endocrine and Breast Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Department of Endocrine and Breast Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Guanglun
authors.initials: G
authors.lastname: Yang
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects; D004965:Estrogen Antagonists / Q000494:pharmacology*; D005260:Female; D005347:Fibroblasts / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D011960:Receptors, Estrogen / Q000502:physiology*; D043562:Receptors, G-Protein-Coupled / Q000502:physiology*; D013629:Tamoxifen / Q000494:pharmacology*; D014162:Transfection
chemical_list: D004965:Estrogen Antagonists; C109584:GPER1 protein, human; D011960:Receptors, Estrogen; D043562:Receptors, G-Protein-Coupled; D013629:Tamoxifen