Atcc crl 1740

Normal prostate epithelial cell line RWPE-1(ATCC^® CRL-11609™), androgen-dependent (LNCaP) and androgen-independent (PC3) human prostate cancer cell lines (ATCC^® CRL-1740™) were obtained from ATCC (Manassas, VA, USA). The cell lines were cultured with modified Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA) and 100 U/mL penicillin and streptomycin at 37 °C with 5% CO₂. The cells were subcultured upon reaching 80–90% confluence. The cells exhibiting logarithmic growth were subsequently seeded into a 6-well plate at 4 × 10⁵ cells per well. LNCaP and PC3 cells were prepared for transfection as per the instruction of the Lipofectamin2000 kit (11668-019, Invitrogen, Carlsbad, CA, USA). Upon reaching 80% confluence, the LNCaP cells were transfected with miR-183 mimic, miR-183 inhibitor, small interfering RNA targeting TPM1 (si-TPM1), TPM1 overexpression plasmid (oe-TPM1) or their negative controls (mimic-NC, inhibitor-NC, si-NC and oe-NC). The PC3 cells were transfected with miR-183 mimic, miR-183 inhibitor, mimic-NC or inhibitor-NC. All aforementioned transfection plasmids were purchased from Shanghai GenePharma Co., Ltd (GenePharma, Shanghai, China).

To complete the evaluation of complexes C1, C3, and C4, in vitro biological behavior was assessed. Two different cell lines derived from prostatic adenocarcinoma were used, LNCaP, which express the AR, and PC3 cells, which do not express it.
The adherent cell line LNCaP (ATCC^® CRL-1740™, American Type Culture Collection, Manassas, VA, USA) was cultured in a conditioned RPMI 1640 medium (Capricorn Scientific, Ebsdorfergrund, Germany) supplemented with 10% fetal bovine serum (Capricorn Scientific), 100 U/mL of penicillin (Sigma-Aldrich, St. Louis, MI, USA), and 100 μg/mL of streptomycin (Sigma-Aldrich) in T75 flasks (Greiner Bio-one, Frickenhausen, Germany) at 37 °C and 5% CO₂. The studies were performed with monolayer cultures with less than 25 passages.
The adherent cell line PC3 (ATCC^® CRL-1435™, American Type Culture Collection, Manassas, VA, USA) was cultured in a DMEM medium (A1316, 9050 PanReac AppliChem, ITW Reagents, Darmstadt, Germany) supplemented with 10% fetal bovine serum, 100 U/mL of penicillin, and 100 μg/mL of streptomycin in T75 flasks (Greiner Bio-one, Kremsmünster, Austria) at 37 °C and 5% CO₂. The studies were performed with monolayer cultures with less than 20 passages.
Cellular uptake, efflux, and binding studies for each complex were performed.

PC3, DU-145 and LNCaP human prostate cancer cell lines were obtained from the American Type Culture Collection (ATCC CRL-1435, ATCC HTB-81 and ATCC CRL-1740, respectively) (American Type Culture Collection, Rockville, MD, USA). The cells were routinely grown in RPMI 1640 medium supplemented with 100 IU/mL penicillin G sodium, 100 µg/mL streptomycin sulfate, 0.25 µg/mL amphotericin B (Invitrogen, Paisley, UK) and 10% fetal bovine serum. All cell lines were incubated at 37 °C in 5% CO₂ and routinely tested for Mycoplasma infection. For treatment experiments, the cells were plated and grown for 24 h, the medium was then replaced with serum-free RPMI 1640 and then incubated with different treatments for the indicated times. The cells were used at passages 4–20.