LNCaP and C4–2 were purchased from ATCC (ATCC.com). LAPC4 and LREX were provided by Dr. Sawyers, Memorial Sloan Kettering Cancer Center.
LNCaP, C4–2, 22Rv1 and LREX cells were cultured in
RPMI1640+GlutaMAX (Gibco, Life Technologies), 10% FBS (Gibco, Life Technologies), 1%
Penicillin/Streptomycin (Corning). LAPC4 cells were cultured in IMDM supplemented with 10% FBS and 1%
Penicillin/Streptomycin. For androgen deprivation
in vitro studies, 10%
charcoal stripped serum (CSS) (Gibco, Life Technologies) was used in place of 10% FBS. All cell lines were tested for mycoplasma contamination (PCR Mycoplasma Detection; primer sequence in
Supplementary Table 1) every 6 months. LuCaP PDX models (provided by Dr. E. Corey and Dr. R. Vessella, Washington University) were grown in organoid culture in advanced DMEM/F12 media with supplements, as previously published (18 (
link)). For
in vitro studies, ipatasertib (Chemietek) and
enzalutamide (Selleckchem) were dissolved in DMSO.
CellTitre Glo Assay (Promega) was used to assess cell viability. For
in vivo studies, ipatasertib (Division of Cancer Treatment and Diagnosis, NCI Developmental Therapeutics Program) and
enzalutamide (DCTD, NCI DTP) were dissolved in 1:1 of
labrasol (Gattefosse) to
PEG400 (Sigma).
Adelaiye-Ogala R., Gryder B.E., Nguyen Y.T., Alilin A.N., Grayson A.R., Bajwa W., Jansson K.H., Beshiri M.L., Agarwal S., Rodriguez-Nieves J.A., Capaldo B., Kelly K, & VanderWeele D.J. (2020). Targeting the PI3K/AKT pathway overcomes enzalutamide resistance by inhibiting induction of the glucocorticoid receptor. Molecular cancer therapeutics, 19(7), 1436-1447.