Nci h1573

Human breast cancer cell lines MCF7 and T47D, human lung adenocarcinoma cell lines NCI-H1573 and NCI-H1437, prostate cancer cell line 22Rv1, and lentivirus packaging HEK293T cell were obtained from the American Type Culture Collection. MCF7 and HEK293T cell lines were maintained in Dulbecco's Modified Eagle Medium (DMEM, Gibco, #11995065) containing 10% fetal bovine serum (Gibco, #10099141) and 1% penicillin-streptomycin (Beyotime, #C0222). T47D and 22Rv1 cell lines were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, #11875500) supplied with 10% fetal bovine serum and 1% penicillin-streptomycin. NCI-H1437 and NCI-H1573 cell lines were propagated in RPMI-1640 supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, 1% GlutaMAX (Gibco, #35050061), 1% Non-Essential Amino Acids (Gibco, #11140050) and 1% Sodium Pyruvate (Gibco, #11360070). All cell lines were grown at 37°C in a humidified atmosphere of 5% CO₂. MCF7 cells were treated with a series concentration of JQ-1(+) for 8 hours followed by RNA extraction and RT-qPCR to determine the expression of DSCAM-AS1.

Human lung cancer NCI-H1573, A549, NCI-H1299, and NCI-H1975 cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). A549 cells were routinely cultured in F12 Kaighn’s medium (Invitrogen, Carlsbad, CA, USA), and NCI-H1573, NCI-H1299, and NCI-H1975 were cultured in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA). The media used for the routine cell culture were supplemented with 10% (v/v) fetal bovine serum (FBS) and a 1% (w/v) combination of penicillin, streptomycin, and neomycin (Invitrogen, Carlsbad, CA, USA). The cultures were incubated in 75 cm² vented culture flasks at 37 °C in 5% CO₂/95% humidified air. The cells were trypsinized after they were 80–90% confluence and seeded onto the appropriate well plates.

Primary CAFs used in the present study were isolated from NSCLC specimens, as previously reported [1 (link)]. The study was conducted in accordance with a protocol approved by the University Health Network (UHN) Research Ethics Board. NSCLC tissues were collected with informed consent from all patients. An overview of CAFs used in this study is presented in Table 1. CAF cells were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Thermo Scientific, Burlington, ON, Canada). All CAF primary cultured cells were maintained and used at early passage (passage 2–5). Murine C2C12 myoblasts stably expressing either human integrin α1 (C2C12-α1), human integrin α2 (C2C12-α2) or human integrin α11 (C2C12-α11) were obtained from Prof. Donald Gullberg (University of Bergen, Norway) and were cultured in DMEM supplemented with 10% FBS. The NCI-H1573, H2073, H358, H1975 and H2009 NSCLC cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA) and were cultured in RPMI 1640 media supplemented with 10% FBS.

Human NSCLC cell lines NCI-H1975, NCI-H1299, NCI-H1437, and NCI-H1573 were purchased from ATCC (American Type Culture Collection, Manassas, Virginia). PC9 and HCC827 cell lines were obtained from Cell Biology and Biotherapy Unit, Istituto Nazionale Tumori di Napoli, IRCCS “G. Pascale.” The EGFR TKI-resistant cell lines, PC9GR and HCC827GR, were generated by culturing the respective parental cell lines in the presence of increasing concentrations of TKI Gefitinib (from 0.05 to 0.5 μM and from 0.1 to 1 μM, respectively) for 8 weeks, to reach a concentration 10 times higher than the initial IC₅₀. Cell lines were cultured in RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum (Gibco), 1% L-glutamine (2 mM, Lonza), 1% streptomycin-penicillin (EuroClone) at 37°C in a 5% CO₂ humidified atmosphere. Short tandem repeat (STR) loci for cell lines authentication were evaluated for all cell lines, by using the GenePrint10-System (Promega).