PC-3, DU145 and LNCaP cell lines (PCa) and RWPE-1 (normal prostate) were purchased from American Type Culture Collection (ATCC). Cells were cultured in a humidified incubator at 37°C with 5% CO2 according to the protocol outlined in a previous study (11 (link)–13 (link)). Human premicroRNA Expression Construct Lenti-miR-505 [PMIRH505PA-1; System Biosciences (SBI), LLC] was used as the pMIRNA1 lentivectors to express miR-505 precursor (pre-505). Scramble control hairpin in pCDH-CMV-MCS-EF1α-copGFP (CD511B-1) was designed as a negative control (pre-NC; cat. no. PMIRH000PA-1; SBI). With pPACKH1 Packaging Plasmid Mix (cat. no. LV500A-1; SBI), pre-505/pre-NC were transfected into 293TN cells (SBI) in order to package the construct. By using the LentiConcentin Virus Precipitation Solution (cat. no. LV810A-1; SBI), the virus particles were collected according to the manufacturer's protocol. Following transfection, PC-3 and LNCaP cells were isolated and then seeded.
NRCAM expression plasmid (pCMV-NRCAM) was obtained from Biogot Technology Co., Ltd. The cells transfected with pCMV-NRCAM or pCMV (empty vector) were used as a corresponding control. The cells were isolated for functional analyses at 48 h following transfection.
NRCAM expression plasmid (pCMV-NRCAM) was obtained from Biogot Technology Co., Ltd. The cells transfected with pCMV-NRCAM or pCMV (empty vector) were used as a corresponding control. The cells were isolated for functional analyses at 48 h following transfection.
