Sk br 3

Human breast cancer cell lines, MDA-MB231, MCF-7, SK-BR-3, and BT-474, were obtained from Korean Cell Line Bank (Seoul, Korea). The normal MCF-10A cells were obtained from American Type Culture Collection (Manassas, VA, USA). The human breast cancer cells were propagated in RPMI-1640 medium supplemented with 10% FBS and 1% antibiotic solution. MCF-10A cells were cultured in DMEM/F-12 medium supplemented with 10% FBS and 1% antibiotic solution. All the cells were maintained at 37 °C in 5% CO₂ conditions.

SK-BR-3 and MCF-7 human breast cancer cell lines were purchased from the Korean Cell Line Bank (Seoul, Korea) and maintained in DMEM containing 10% fetal bovine serum and 1% antibiotic/antimycotic solution. Media for MCF-7 cells was also supplemented with insulin at 10 μg/mL.

CAFs were isolated as previously described.^{45 (link)} Briefly, tissue from early-stage IDC (stage 1) that was less than 10 mm in diameter was sliced and then digested overnight with a collagenase preparation (ISU ABXIS; Seoul, South Korea). Digested tissue was filtered through a 70 μm cell strainer (SPL Life Science; Pocheon-si, South Korea). Cells were separated by Ficoll gradients, washed with PBS, resuspended with DMEM/F12 cell culture medium containing 20% (v/v) fetal bovine serum (FBS), 100 IU/ml penicillin, and 100 μg/ml streptomycin (Gibco BRL; Grand Island, NY, USA) and cultured at 37 °C in a humidified incubator containing 5% CO₂. The fibrotic nature of the isolated cells was confirmed by microscopic determination of morphology and immunofluorescence characterization using the antibodies against vimentin (Abcam; Cambridge, UK), cytokeratin (Dako; Glostrup, Denmark) and cytokeratin 5 (Novocastra; Newcastle upon Tyne, UK). Breast cancer cell lines (BT-474, MCF7, SK-BR-3, MDA-MB-231) were purchased from Korean Cell Line Bank (Seoul, South Korea) (authenticated using morphology and STR profiling) and cultured with DMEM cell culture medium containing 10% (v/v) fetal bovine serum (FBS), 100 IU/ml penicillin, and 100 μg/ml streptomycin at 37 °C in a humidified incubator containing 5% CO₂.

The human BRC cell lines BT549, BT20, T47D, ZR75-1, SKBR3, BT474, and MDA-MB453 were purchased from the Korean Cell Line Bank. Other cell lines (MDA-MB231 and MCF7) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells from the ATCC were certified by the results of a short tandem repeat (STR) DNA profiling assay, a cytochrome C oxidase I assay, and a mycoplasma contamination assay. The nine BRC cell lines were characterized by the short tandem repeat (STR)-polymerase chain reaction (PCR) method and screened for mycoplasma contamination.
The BRC cells used in this study were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS, HyClone) and 1% penicillin/streptomycin (HyClone) (hereafter referred to as complete RPMI1640 medium) at 37 °C in 5% CO₂.

MCF10A, ZR75-30, MDA-MB-453, BT20, JIMT-1, MDA-MB-231, and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). MCF7, T47D, ZR75-1, SKBR3, BT474, HCC-1954, HCC1419, and HS578T cells were purchased from the Korean Cell Line Bank. MCF7, T47D, MDA-MB-231, and HEK293T cells were cultured in DMEM (Welgene, Daegu, Republic of Korea) supplemented with 10% FBS, and SKBR3, BT474, HCC-1954, HCC-1419, JIMT-1, and HS5788T cells were cultured in RPMI (Welgene) supplemented with 10% FBS at 37 °C in a 5% CO2 atmosphere. Trastuzumab was obtained from Roche (Basel, Switzerland). The β-Catenin/TCF inhibitor FH-535 was purchased from Sigma-Aldrich (St. Louis, MO, USA).

Cell lines were obtained from ATCC (USA) (MKN-28, AGS, KATOIII, HCT-116, MDA-231, SK-BR-3, JIMT-1, U87MG, A549, CEM-119, HeLa, Jurkat and HDF) and from Korean Cell Line Bank (Seoul National University, Korea) (SNU-216, SNU-638, SNU-719, and SNU-C5) were cultured with designated media supplemented with 10% fetal bovine serum and 1x penicillin-streptomycin at 37 °C 5% CO₂ incubator. Human stomach cancer tissues and peri-tumoral normal counter parts were excised within 10 min from the gastrectomy, and preserved in RNAlater solution until RNA purification at 4 °C. All patients provided informed consent prior to collection of tissues, and all methods were performed in accordance with the relevant guidelines and regulations approved by the Institutional Review Board of National Cancer Center of Korea (NCCNCS13732).

Lung (NCI-H23, A549, NCI-H358, Calu-1, NCI-H460, and NCI-H1299), breast (SK-BR-3, MCF-7, and MDA-MB-231), colorectal (HCT116), hepatocellular (SK-HEP-1), and cervical (HeLa) cancer cells were obtained from the Korean Cell Line Bank (Seoul, Korea). Melanoma cell lines (A375 and A2058) were obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were cultured in 10% fetal bovine serum (FBS) as well as penicillin/streptomycin-contained Dulbecco’s modified Eagle’s medium (DMEM) and Roswell Park Memorial Institute (RPMI) 1640 at both 20% O2 and 5% CO2. Cell culture medium, FBS, and antibiotics were purchased from HyClone Thermo Scientific (Waltham, MA, USA). Simvastatin (S6196), Lovastatin (438185), Lonafarnib (SML1457), and GGTI-2133 (G5294) were purchased from Sigma Aldrich (St. Louis, MO, USA). Atorvastatin (S5715), Fluvastatin (S2061), and MK-2206 (S1078) were obtained from Selleckchem (Houston, TX, USA). All chemicals stock solution was dissolved in dimethyl sulfoxide (DMSO).