Mda mb 231

HCT116, MDA-MB-231, A549, and AsPC-1 were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan). The human bone marrow stromal cell line HS-5 was kindly provided by Prof. Yu, Alice Lin-Tsing (Genomics Research Center, Academia Sinica, Taipei, Taiwan). The cells were cultured in Roswell Park Memorial Institute medium (RPMI)1640 (HCT116, MDA-MB-231 and AsPC-1) or Dulbecco's Modified Eagle's medium (DMEM) (A549 and HS-5) respectively supplemented with 10% (v/v) heat-inactivated fetal bovine serum (both from Invitrogen^TM Life Technologies, Carlsbad, CA, USA), 100 U/mL of penicillin, and 100 μg/mL of streptomycin (Biological Industries, Kibbutz Beit Haemek, Israel). All cells were maintained at 37°C in a humidified atmosphere of 5% CO₂ in air.

Women with Breast Imaging Reporting and Data System (BI-RADS) category 4 or 5 due to suspicious malignant MC on screening or diagnostic mammography were recommended for stereotactic breast biopsy. Biopsies of breast MC and adjacent normal tissue (non-MC) were obtained by the stereotactic 10-gauge vacuum-assisted breast biopsy system (Vacora, Bard Medical Systems, Tempe, AZ). In general, 15 to 20 specimens per lesion were obtained. Women were included if mammography between January 2010 and December 2011 showed that MC specimens were insufficient for clinical pathology diagnosis.
Breast cancer cell lines, MCF-7 (BCRC#60436) and MDA-MB-231 (BCRC#60549) from the Bioresource Collection and Research Center (BRCR, Hsinchu, Taiwan) were cultured in DMEM supplemented with 10% fetal bovine serum (Invitrogen) in a humidified atmosphere of 95% air and 5% CO₂ at 37°C.

The MDA-MB-231 and MCF7 breast cancer cells, purchased from Bioresource Collection and Research Center (BCRC, Taiwan), were cultured in Roswell Park Memorial Institute (RPMI)-1,640 medium (Biowest) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C and 5% CO_2. Before live-cell monitoring, the cells were transfected with an actin–green fluorescent protein (GFP) reporter using a baculovirus expression vector (CellLight^® Actin-GFP BacMan 2.0; Invitrogen) at 70% confluence with a particle per cell ratio of 30 and then incubated at 37°C and 5% CO₂ for 24 h.

The ts-v-Src RK3E cells were created and cultured as previously described [12 (link)]. H-1299, HT-29 and B16F10 cells (American Type Culture Collection; Manassas, USA) were obtained from Dr. Chao-Hsiung Lin, National Yang-Ming University, Taiwan). A2058, K562 and MDA-MB-231 cells were purchased from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). BJ-1 cells [13 (link)] were from Dr. Yeu Su at National Yang-Ming University, Taiwan. H-1299 and HT-29 cells were cultured in RPMI medium (Gibco, Grand Island, NY) containing 10% fetal bovine serum (FBS, Biological Industries, Kibbutz Beit Haemek, Israel), and 1% penicillin-streptomycin-glutamine (PSG). K562 cells were cultured in RPMI medium supplemented with 10% FBS, 1% PSG, and 1% sodium pyruvate (Gibco, Grand Island, NY). B16F10, A2058 and BJ-1 cells were cultured in DMEM medium (Gibco, Grand Island, NY) containing 10% FBS and 1% PSG. MDA-MB-231 cells were cultured in DMEM medium containing 10% FBS, 1% PSG, and 1% sodium pyruvate. All cells were incubated in a 5% humidified incubator at 35°C.

Two human breast carcinoma cell lines (MDA-MB-231 and MCF-7) were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan). The human mammary gland epithelial cell line (MCF-10A) and human umbilical vein vascular endothelial cell (HUVECs) line were purchased from the American Type Culture Collection (VA, USA). HUVECs were grown under standard conditions in endothelial cell growth medium supplemented with the EGM-2 Bullet Kit. HUVECs at passages 4-8 were used for all experiments. The cancer cells were grown in F-12K medium containing 10% fetal bovine serum. Cells were maintained at 37ºC in a humidified atmosphere with 5% CO₂. Artemisinin and vinorelbine were purchased from Sigma Chemical Co. (MO, USA).

The human breast cancer cell lines MDA-MB-231 and MCF-7 were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan), and cultured in its standard medium as recommended by the BCRC. Briefly, MDA-MB-231 was cultured in Leibovitz’s L-15 medium with 2 mM l-glutamine and 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere without CO₂. MCF-7 was cultured in a minimum essential medium Eagle with 0.1 mM non-essential amino acids, 1 mM sodium pyruvate and 10% FBS at 37 °C in a humidified atmosphere with 5% CO₂. Culture medium, FBS, and cultured supplements were all purchased from Invitrogen (Carlsbad, CA, USA). Cell lines were authenticated annually by short-tandem repeat analysis and routinely tested for mycoplasma contamination (BCRC).

MDA-MB-231 and MKN-45 cell lines were obtained from the Bioresource Collection and Research Center in Taiwan and Japanese Collection of Research Bioresources Cell Bank, respectively. HCT116 and Colo205 cell lines were obtained as a gift from Dr. Chiung-Tong Chen, National Health Research Institute, Taiwan. The cells were cultivated in DMEM, RPMI1640, and McCoy’s 5A supplemented with 10% fetal bovine serum (FBS) and 250 units/ml of penicillin/streptomycin solution. All cell culture media and supplements were purchased from Gibco (Carlsbad, CA, USA). The cells were maintained in the presence of 5% CO₂ at 37 °C.