Breast tumor tissues and matched adjacent normal tissues were obtained from patients who were diagnosed with breast cancer and who had undergone surgery at The Second Affiliated Hospital of Shantou University Medical College. Tissue samples were freshly frozen in dry ice and stored at -80 °C until RNA extraction. The study was approved by the Institutional Ethics Committee of the Second Affiliated Hospital of Shantou University Medical College. All research was performed in compliance with government policies and the Helsinki Declaration. Experiments were undertaken with the understanding and written consent of each subject.

Human breast cancer cell lines and human embryonic kidney cell line HEK293T were cultured in Dulbecco's modified Eagle medium (DMEM) (Biological Industries) or Roswell Park Memorial Institute 1640 (RPMI 1640) (Biological Industries). All medium was supplied with 10% fetal bovine serum (FBS) (Biological Industries) and 1% penicillin/streptomycin (Biological Industries) for cell culture. When treating cells with estrogen, phenol-free medium supplemented with 5% charcoal-treated FBS was used.

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