Pc3 prostate cancer cell line

Manufactured by American Type Culture Collection

Sourced in United States

The PC3 prostate cancer cell line is a well-established in vitro model derived from a human prostate adenocarcinoma. It is widely used in cancer research to study the biology and behavior of prostate cancer cells.

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10 protocols using pc3 prostate cancer cell line

Culturing PC3 Prostate Cancer Cells

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PC3 prostate cancer cell line was purchased from ATCC and cultured as per ATCC recommendations.

Komaragiri S.K., Bostanthirige D.H., Morton D.J., Patel D., Joshi J., Upadhyay S, & Chaudhary J. (2016). ID4 promotes AR expression and blocks tumorigenicity of PC3 prostate cancer cells. Biochemical and biophysical research communications, 478(1), 60-66.

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Engineered Cell Lines for Cancer Research

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SKOV3 ovarian cancer cell line, PC3 prostate cancer cell line, and 293T were obtained from ATCC. NCI-H292 lung cancer cell line was kindly provided by Dr. Ite Laird-Offringa. The PC3 cell line was transduced with viral vector to express a high level of PD-L1 on the cell surface as PC3-PD-L1 and sorted to 99% purity. The SKOV3, NCI-H292, and PC3-PD-L1 cell lines were transduced with a lentiviral vector FUW-CD19. The transduced cells were then stained with anti-human CD19 antibody and sorted to 99% purity. The cell culture methods for 293T, engineered SKOV3-CD19, and H292-CD19 were described in our previous publications.¹³ (link)^,⁴⁷ (link) PC3 and PC3-CD19 were maintained in D10 medium consisting of DMEM supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin.

Chen X., Yang S., Li S., Qu Y., Wang H.Y., Liu J., Dunn Z.S., Cinay G.E., MacMullan M.A., Hu F., Zhang X, & Wang P. (2021). Secretion of bispecific protein of anti-PD-1 fused with TGF-β trap enhances antitumor efficacy of CAR-T cell therapy. Molecular Therapy Oncolytics, 21, 144-157.

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Cell Culture Protocols for Cancer Research

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Spodoptera frugiperda (Sf9) insect cells (Expression Systems) were grown in ESF 921 serum-free medium (Expression Systems) at 27 °C and 120 rpm. HEK293T cells were purchased from American Type Culture Collection (ATCC), cultured in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies), supplemented with 10% fetal bovine serum (FBS; Gibco), and maintained in a humidified chamber with 5% CO₂ at 37 °C. The PC3 prostate cancer cell line (ATCC) was cultured in F12 medium supplemented with 10% FBS and maintained in a humidified chamber with 5% CO₂ at 37 °C. 22Rv1 and LNCaP human prostate cancer cells (ATCC) were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium supplemented with 10% FBS and maintained in a humidified atmosphere containing 5% CO₂ at 37 °C.

Cong Z., Zhou F., Zhang C., Zou X., Zhang H., Wang Y., Zhou Q., Cai X., Liu Q., Li J., Shao L., Mao C., Wang X., Wu J., Xia T., Zhao L.H., Jiang H., Zhang Y., Xu H.E., Cheng X., Yang D, & Wang M.W. (2021). Constitutive signal bias mediated by the human GHRHR splice variant 1. Proceedings of the National Academy of Sciences of the United States of America, 118(40), e2106606118.

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Prostate Cancer Cell Line Characterization

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The PC-3 prostate cancer cell line was purchased from ATCC (Manassas, VA, USA). The passage numbers of cells used in experiments ranged from 5 to 20. Thapsigargin (10522) (TG), fura-2-acetoxymethyl ester (14591) (Fura-2), and trans-resveratrol (70675) (RES) were purchased from Cayman Chemical (Ann Arbor, MI, USA). U-73122 (J62898) and 2-Aminoethyl diphenylborinate (A16606) (2-APB) were purchased from Alfa Aesar (Ward Hill, MA, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (AC158990050) (MTT) was purchased from Acros Organics (Morris Plains, NJ, USA). 4′-Butyrate resveratrol (BuRV), 4′-isobutyrate resveratrol (IsoRV) and 4′-pivalate resveratrol (PIV, Fig. 1) were synthesized by the Andrus lab in the Department of Chemistry and Biochemistry at Brigham Young University [29 (link)]. The purity of the compounds was determined to be greater than 98% by NMR and mass spectrometry.Fig. 1

Structures.

Structures of (A) trans-resveratrol, (B) 4′-pivalate trans-resveratrol, (C) 4′-isobutyrate trans-resveratrol, and (D) 4′-butyrate trans-resveratrol.

Fig. 1

Peterson J.A., Crowther C.M., Andrus M.B, & Kenealey J.D. (2019). Resveratrol derivatives increase cytosolic calcium by inhibiting plasma membrane ATPase and inducing calcium release from the endoplasmic reticulum in prostate cancer cells. Biochemistry and Biophysics Reports, 19, 100667.

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Generating Prostate Cancer Cell Lines

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PC3 prostate cancer cell line was purchased from ATCC and cultured as per ATCC recommendations. ID4 was stably silenced in PC3 cells using gene specific shRNA retroviral vector as described earlier [28 (link)].
Mutagenesis of p53: The retroviral p53 vector previously described [28 (link)] was manipulated via mutation induced by site-directed mutagenesis mimicking DU145 cells that harbor mutant p53 (P223L and V274F) [51 (link)] via QuickChange II Site-Directed Mutagenesis kit (Agilent Technologies, USA). Hotspot mutations of p53 (R273H and R175H) [52 (link)] were purchased from Addgene, Inc: pCMV-Neo-Bam p53 R175H (Addgene plasmid #16436) and pCMV-Neo-Bam p53 R273H (Addgene plasmid # 16439) [53 (link)] was a gift from Bert Vogelstein. The following primers were used for generating the P223L and V274F mutants:

Morton D.J., Patel D., Joshi J., Hunt A., Knowell A.E, & Chaudhary J. (2016). ID4 regulates transcriptional activity of wild type and mutant p53 via K373 acetylation. Oncotarget, 8(2), 2536-2549.

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Culturing PC-3 Prostate Cancer Cells

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PC-3 prostate cancer cell line was purchased from American Type Culture Collection (ATCC, Manassas, VA). Cells were grown and maintained at 37 °C, 5% CO₂ in RPMI1640 containing 10% FBS (fetal bovine serum) or in a keratinocyte serum-free medium (K-SFM) supplemented with bovine pituitary extract (BPE) and recombinant epidermal growth factor (EGF) for prostasphere growth (Life Technologies, Gaithersburg, MD).

Zhou Y., Yang J., Zhang R, & Kopeček J. (2014). Combination therapy of prostate cancer with HPMA copolymer conjugates containing PI3K/mTOR inhibitor and docetaxel. European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V, 89, 107-115.

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Culturing PC3 Prostate Cancer Cells

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The PC3 prostate cancer cell line was purchased from American Type Culture Collection (ATCC, Rockville, MD, USA). The cells were maintained at 37°C in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) in a 5% CO₂ incubator, following the instructions provided by ATCC.

Sun H., Zhang L., Shi C., Hu P., Yan W., Wang Z., Duan Q., Lu F., Qin L., Lu T., Xiao J., Wang Y., Zhu F, & Shao C. (2015). TOPK is highly expressed in circulating tumor cells, enabling metastasis of prostate cancer. Oncotarget, 6(14), 12392-12404.

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Prostate Cancer Cell Line Culture

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The PC-3 prostate cancer cell line was purchased from American Type Culture Collection and the LNCaP prostate cancer cell line was purchased from UROCOR (Oklahoma City, OK). Cell lines were cultured in RPMI media plus 5% fetal calf serum (FCS, Life Technologies) and maintained at 37°C under atmospheric oxygen conditions with 5% CO₂ unless noted otherwise.

Wiese A.K., Prior S., Chambers T.C, & Higuchi M. (2017). Intracellular Oxygen Concentration Determined By Mitochondrial Respiration Regulates Production of Reactive Oxygen Species. Integrative cancer biology & research, 1(1), 006.

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Expansion and Characterization of GBM Stem Cells

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GSCs were derived from surgical specimens obtained from GBM patients at the Massachusetts General Hospital (MGH; provided by Dr. Hiroaki Wakimoto) or The Ohio State University James Comprehensive Cancer Center (provided by Dr. Ichiro Nakano), under the appropriate Institutional Review Board approval and were previously characterized^{24 (link),54 (link)}. Cells were expanded as neurospheres and maintained in DMEM/F12 supplemented with B27 minus vitamin A (Life Technologies), heparin (Sigma–Aldrich), 20 ng/mL human recombinant EGF, and bFGF-2 (Peprotech). PC3 prostate cancer cell line was obtained from American Type Culture Collection (ATCC) and grown in DMEM with 10% fetal bovine serum. Cells were maintained at 37 °C in humidified 5% CO₂ incubators.

da Hora C.C., Pinkham K., Carvalho L., Zinter M., Tabet E., Nakano I., Tannous B.A, & Badr C.E. (2019). Sustained NF-κB-STAT3 signaling promotes resistance to Smac mimetics in Glioma stem-like cells but creates a vulnerability to EZH2 inhibition. Cell Death Discovery, 5, 72.

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Prostate Cell Line Culture Techniques

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PC-3 prostate cancer cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), cultured at 37 °C and 5% CO₂ in Ham’s F-12 K (Kaighn’s) basal medium (Gibco Laboratories, Grand Island, NY, USA), supplemented with 10% FBS (Gibco Laboratories) and 1% of 100 U/mL penicillin/streptomycin (Gibco Laboratories). The PNT2 (European Collection of Authenticated Cell Cultures, ECACC, UK) human prostate cell line was purchased from Sigma-Aldrich. The cells were cultured at 37 °C and 5% CO₂ in RPMI 1640 (Gibco Laboratories), supplemented with 10% FBS (Gibco Laboratories), 1% of 100 U/mL penicillin/streptomycin (Gibco Laboratories), and 2% L-glutamine (Gibco Laboratories).

Fassi E.M., Garofalo M., Sgrignani J., Dei Cas M., Mori M., Roda G., Cavalli A, & Grazioso G. (2022). Focused Design of Novel Cyclic Peptides Endowed with GABARAP-Inhibiting Activity. International Journal of Molecular Sciences, 23(9), 5070.

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