Bph 1

The 1E8 and 2B4 cells were derived from the PC-3M human prostate carcinoma cell line. 1E8 was highly metastatic, whereas 2B4 was non-metastatic [26] (link). 22RV1 prostate cancer cell line and BPH1 were purchased from American Type Culture Collection. All cells were cultured in RPMI 1640 (GIBCO, Grand Island, NY, USA) supplemented with 10% fetal bovine serum in a humidified atmosphere containing 5% CO₂ at 37°C.

Human PCa cell lines (LNCaP, DU145, PC3), human prostatic hyperplasia cells (BPH-1), transgenic adenocarcinoma mouse prostate cell (TRAMP-C2), and prostatic epithelial cell line (RWPE-1) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The above cells were supplemented with RPMI 1640 medium containing 10% fetal bovine serum (Gibco, USA) at 37°C, and the medium was changed once a day. Has-miR-145-5p mimics, mimic-NC, si-WIP1, si-NC, vector-NC, and vector WIP1 were purchased from Guangzhou Ruibo Biology Co., Ltd. When the cells were cultured to 60% fusion degree, the Lipofectamine^TM 2000 (Life Technologies, USA) specification was used for transfection of siRNA (50 nmol/L) with liposome Lipofectamine^TM2000. After 24 h of transfection, the cells were collected for cell proliferation activity detection.

Human normal prostatic epithelial (RWPE-1) and human BPH (BPH-1) cell lines were obtained from American Type Culture Collection (Manassas, VA, USA). Cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (penicillin (50 U/mL) and streptomycin (50 μg/mL)) in an atmosphere of 5% CO₂ at 37 °C.

Human prostate cancer cell lines LNCaP, C4-2, C4-2B, CWR22Rv1, PC-3, DU145, and human benign prostatic hyperplasia cell (BPH1) and normal prostatic epithelial cell (RWPE1) were purchased from American Type Culture Collection (Manassas, VA, USA) and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Gibco, NY, USA) at 37°C, in humidified air containing 5% of CO₂. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was purchased from Sigma-Aldrich Co. (MO, USA). Rabbit anti-CD82 (Cat. No. ab109529) and rabbit anti-ADAM17 (Cat. No. ab173579) monoclonal antibodies were purchased from Abcam (MA, USA), and mouse anticytoplasmic fragments of E-cadherin monoclonal antibody were from BD Biosciences (Cat. No. 610181, CA, USA), and rabbit anti-HA (Cat. No. 3724S) and rabbit anti-Flag (Cat. No. 14793S) monoclonal antibodies were from Cell Signaling Technology, Inc. (MA, USA). Mouse anti-Flag monoclonal antibody (Cat. No. F1804) was from Sigma-Aldrich Co. (MO, USA). Mouse anti-HA monoclonal antibody (Cat. No. 26183) was from Thermo Fisher Scientific Inc. (MA, USA). Mouse anti-GAPDH monoclonal antibody was purchased from Kangchen Bio-tech Inc. (Cat. No. KC-5G4, Shanghai, China).

All human cell lines were purchased from the American Type Culture Collection (ATCC, USA), including DU145, PC3, and BPH-1. All cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, USA; catalog number: C11875500BT) supplemented with 10% fetal bovine serum (FBS; Gibco, USA; Cat.10270–106), 0.1 mg/mL streptomycin, and 100 U/mL penicillin (Gibco, USA; catalog number: 15,140–122) and were maintained in a humidified incubator at 37°C containing 5% CO₂. Total RNA was obtained with the RNeasy mini kit (QIAGEN, Germany, Cat. No. 74,104) and reverse transcribed with the RT kit (TaKaRa, Japan, Cat. No. NR037A). The cDNA products were then subjected to real-time PCR using Fast SYBR® Green Master Mix (Life technology, USA; Cat. No: 4,385,610). The sequences of all primers used for PCR were documented in the supplementary materials.

Three prostate cell lines (LNCaP, PC-3 and BPH-1) were obtained from American Type Culture Collection (Rockville, MD, USA). Cells were cultured in RPMI-1640 media (Invitrogen, Carlsbad, CA, USA) supplemented with 100 IU/mL penicillin G sodium, 100 mg/mL streptomycin sulfate and 10 % (v/v) fetal bovine serum (Hyclone, Logan, UT, USA). Cultures were maintainedat 37 °C in a 5 % CO₂ environment.