MCF10A (Human breast epithelial cells, ATCC, Manassas, VA, USA), MCF-7, and MDA-MB-231 (BC cells, Cell Bank of Chinese Academy of Science, Shanghai, China) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS, 100 units/mL penicillin and 100 mg/mL streptomycin (Invitrogen, Carlsbad, CA, USA) under condition of 5% CO2 flow at 37°C. DOX‐resistant MCF‐7 (MCF‐7/DOX) and DOX‐resistant MDA-MB-231 (MDA-MB-231/DOX) cells were generated by continuous exposure of MCF‐7 or MDA-MB-231 cells to gradient concentrations of DOX (Sigma‐Aldrich, St Louis, MO, USA) [19 (link)]. In addition, in this study, 1.5 μg/mL DOX was applied for the treatment of BC cells, and 30 μg/mL DOX was used to treat DOX-resistant BC cells.
Mda mb 231
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Italy, Israel, Switzerland, Ireland
MDA-MB-231 is a well-characterized cell line derived from a human breast adenocarcinoma. It is commonly used in cancer research and drug development studies.
Automatically generated - may contain errors
Lab products found in correlation
Mda mb 231, by American Type Culture Collection (112 mentions)
Mcf 7, by Merck Group (74 mentions)
Mcf 7, by American Type Culture Collection (60 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (42 mentions)
Fetal bovine serum (fbs), by Merck Group (40 mentions)
Penicillin streptomycin, by Merck Group (32 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (31 mentions)
Rpmi 1640, by Merck Group (26 mentions)
Mda mb 468, by Merck Group (26 mentions)
Mcf 10a, by American Type Culture Collection (24 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (23 mentions)
Mda mb 468, by American Type Culture Collection (21 mentions)
L glutamine, by Merck Group (20 mentions)
Rpmi 1640 medium, by Merck Group (17 mentions)
Penicillin, by Merck Group (16 mentions)
Streptomycin, by Merck Group (15 mentions)
Skbr3, by American Type Culture Collection (15 mentions)
Bt549, by American Type Culture Collection (15 mentions)
Bt 549, by Merck Group (14 mentions)
Sk br 3, by Merck Group (13 mentions)
213 protocols using mda mb 231
1
Breast Cancer Cell Line Maintenance and Treatment
2
Cultivation and Characterization of Breast Cancer Cell Lines
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The cell lines were cultivated at 37 °C in 5% CO2 and 90% humidity under sterile conditions. The two human breast cancer cell lines MCF-7 (MCF-7; Merck KGaA, Darmstadt, Germany) and MDA-MB-231 (MDA-MB-231; Merck KGaA, Darmstadt, Germany) were grown in Dulbecco’s modified Eagle’s medium (DMEM; PAN-Biotech GmbH, Aidenbach, Germany) supplemented with 10% fetal bovine serum (FBS; Biochrome AG, Berlin, Germany), 1% sodium pyruvate, 2 mM glutamine, 100 U/mL of penicillin and 100 µg/mL. MCF-7 cells (ER positive; PgR positive; p53wt) are deficient of caspase-3, while MDA-MB-231 are triple negative, i.e., ER negative; PgR negtaive; p53 mutated, and caspase-3 intact [78 (link),79 (link),80 (link),81 (link)]. Both splitting and harvesting the cells was conducted by trypsination (10% trypsin in DPBS) on a heating plate.
3
Establishing Diverse Cell Line Panel
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The following tumour cell lines were purchased from ECACC directly from Sigma-Aldrich:breast: MDA-MB-231[16 (link)](92020424), MCF7[17 (link)] (86012803); colon: HT29[18 (link)] (91072201), SW480[18 (link)] (87092801), SW620[18 (link)] (87051203); lung: A549[19 (link)] (86012804) and prostate: PC-3[20 (link)] (90112714); as well as endothelial cells: HUVEC (S200-05N) and leucocytes: Jurkat cells[21 (link)] (88042803).The brest cancer tumour cell HCC1937 lines[22 (link)](ATCC-CCL-247)and HCC1954[23 (link)] (ATCC CRL-2338) were purchased from ATCC; the endometrial cell lines HEC1A and HEC1A stably expressing the ETV5 transcription factor (HEC1A-ETV5) were previously described [24 (link)–26 (link)]. Medium for cell culture were obtained from Gibco (DMEM: SW480, SW620, A549, MCF7, MDA-MB-231, PC-3; McCoyy᾿s: HT29, HEC1A, HEC1A-ETV5; RPMI 1640: HCC1937, HCC1954,Jurkat), and from Lonza (Basel, Switzerland) (EGM-2: Huvec). The medium was supplemented with 10% FBS and 1% streptavidin-penicillin (Life Technologies, Carlsbad Ca, USA). Cells were maintained at 37°C in a 5% CO2 incubator. For selected experiments, we transduced MDA-MB-231 and SW480 cells with lentiviral transduction particles (turboGFP, Sigma-Aldrich) to establish cells that expressed green fluorescent protein (GFP); cells were further selected with puromycin (5 μg/mL) and checked by flow cytometry (96%).
4
Establishing Paclitaxel-Resistant Breast Cancer Cells
5
TLR4 Activation in Human Tumor Cell Lines
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The human tumor cell lines SW480 (primary colon adenocarcinoma), SW620 (metastatic colon adenocarcinoma), MDA-MB-231 (metastatic breast adenocarcinoma) and U87-MG (glioblastoma) were purchased from Sigma-Aldrich. Cells were grown in two different Dulbecco’s modified essential media (DMEM high glucose for SW480 and SW620 and DMEM low glucose for MDA-MB-231 and U87-MG) (Sigma-Aldrich) supplemented with 10% FBS (Gibco) and 1% penicillin/streptomycin solution (Gibco) at 37 °C with 5% CO2 in humidified air.
To activate TLR4 cells were incubated with [1 µg/ml] LPS (E. coli 055:B5 LPS, Sigma-Aldrich) for 24 hours, washed three times with PBS and then culture medium was replaced with fresh medium supplemented with 10% certified exosomes-free serum (Gibco). After 24 hours, supernatants were collected and stored at −20 °C until use, while cells were harvested, and their proliferation/viability was determined by the trypan blue exclusion assay.
Cell homogenates were analyzed for TLR4 expression by immunoblotting and the human anti-TLR4 (1:500, Cell Signaling) and anti-β-actin (1:10,000, Sigma-Aldrich) were used as primary antibodies.
All methods of analysis were carried out in accordance with the relevant guidelines and regulation with appropriate quality control.
To activate TLR4 cells were incubated with [1 µg/ml] LPS (E. coli 055:B5 LPS, Sigma-Aldrich) for 24 hours, washed three times with PBS and then culture medium was replaced with fresh medium supplemented with 10% certified exosomes-free serum (Gibco). After 24 hours, supernatants were collected and stored at −20 °C until use, while cells were harvested, and their proliferation/viability was determined by the trypan blue exclusion assay.
Cell homogenates were analyzed for TLR4 expression by immunoblotting and the human anti-TLR4 (1:500, Cell Signaling) and anti-β-actin (1:10,000, Sigma-Aldrich) were used as primary antibodies.
All methods of analysis were carried out in accordance with the relevant guidelines and regulation with appropriate quality control.
6
Establishment of Cisplatin-Resistant Breast Cancer Cell Lines
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MCF-7 and MDA-MB-231 cells were purchased from ATCC. In brief, the parental BC cells (defined MCF-7/Par and MDA-MB-231/Par) were continuously exposed to an ascending series of DDP (0.1~10 μM, Sigma-Aldrich, MO, USA) to induce DDP-resistant cells (MCF-7/DDP and MDA-MB-231/DDP). All cells were maintained in 10% FBS-supplemented DMEM containing 1% penicillin-streptomycin for further use.
7
Breast Cancer Cell Line Culturing
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The breast cancer cell lines MDA-MB-231 and 4T1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were grown in IMDM (MDA-MB-231, Cytogen, GmbH Bienenweg, Berlin, Germany) or RPMI 1640 w/Gluta Max (4T1, Gibco, Grand Island, NY, USA). Mediums were supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA), 1% antibiotics solution (10,000 U/mL of penicillin and 10 mg/mL of streptomycin, 25 µg/mL of amphotericin B (only for MDA-MB-231 cells) (Sigma Aldrich, St. Louis, MO, USA)), and 1% nonessential amino acids (only for MDA-MB-231, Sigma Aldrich). To detect mycoplasma contamination, MycoAlert Assay Control Set, (Lonza, Verviers, Belgium) was used. 5-FU was obtained from Sigma Aldrich. ISC was synthesized by the Centre of Molecular and Macromolecular Studies Polish Academy of Sciences, Division of Organic Chemistry [13 (link)].
8
Cell Line Cultivation and Maintenance
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MCF-7, MDA-MB-231, PC-3, DU145 and LNCaP cells were obtained from ATCC (Rockville, MD, USA). MDA-MB-231 cells overexpressing ERα (MDA-MB-231-ERα) were kindly provided by Dr George Reid (EMBL-Heidelberg). MCF-7, MDA-MB-231, MDA-MB-231 ERα, PC-3 and DU145 were maintained in DMEM medium (Sigma Aldrich, St Louis, MO, USA) supplemented with 10% fetal bovine serum (Life Technologies/Invitrogen, Carlsbad, CA, USA), 100 U/ml penicillin and 100 μg/ml streptomycin (Sigma Aldrich). LNCaP cells were maintained in RPMI medium (Sigma Aldrich) supplemented with 10% fetal bovine serum (FBS) (Life Technologies/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin (Sigma Aldrich). Clonetics Primary Prostate Epithelial Cells (PrEC) were obtained from Lonza (Walkersville, MD, USA) and maintained in Prostate Epithelial Cell Medium BulletKit, Clonetics, PrEGM, BulletKit (CC-3166) containing the following growth supplements: bovine pituitary extract, Hydrocortisone, human epidermal growth factor, Epinephrine, Transferrin, Insulin, Retinoic Acid, Triiodothyronine, and GA-1000. All cells were cultured at 37 °C and 5% CO2.
9
Culturing and Steroid Hormone Depletion of Breast Cancer Cell Lines
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Human breast cancer MCF-7 cells were obtained from RIKEN BioResource Center (Tsukuba, Japan). SK-BR-3 and MDA-MB-231 cells were purchased from American Type Culture Collection (ATCC, Rockville, MD). The MCF-7, SK-BR-3 and MDA-MB-231 cells were grown in minimal essential medium (MEM, Sigma, St. Louis, MO), McCoy's 5A (Thermo Scientific, Waltham, MA) and Leibovitz's L-15 medium (ATCC) supplemented with 10% fetal bovine serum (FBS), respectively. MCF-7 and SK-BR-3 cells were maintained at 37°C with 95% air and 5% CO2. MDA-MB-231 cells were maintained at 37°C 100% air without CO2.
For removal of steroid hormones from FBS, a 5% charcoal and 0.5% dextran (Sigma) suspension in FBS was incubated at 37°C for 1 hr with shaking. The suspension mixture was then centrifuged at 2,500 rpm for 20 min, and the supernatant was filtered through a 0.2 μm filter.
For removal of steroid hormones from FBS, a 5% charcoal and 0.5% dextran (Sigma) suspension in FBS was incubated at 37°C for 1 hr with shaking. The suspension mixture was then centrifuged at 2,500 rpm for 20 min, and the supernatant was filtered through a 0.2 μm filter.
10
In Vitro Cytotoxicity Evaluation of Gold Nanoparticles
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The triple-negative human breast adenocarcinoma cell line MDA-MB-231 (American Type Culture Collection [ATCC], Manassas, VA, USA) was used for cellular PDT tests. MDA-MB-231 cells have been reported for their resistance to traditional chemotherapies, aggressive pathological features, and high rates of metastasis and recurrence, and stand as a suitable in vitro model for evaluating PDT efficiency.28 (link),29 (link) MDA-MB-231 cells were cultured in Leibovitz L-15 Medium (Sigma-Aldrich Co.) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin in a humidified incubator at 37°C without CO2. Media were refreshed every 2 days and the cells were passaged at a confluence of 70%–80%. For cytotoxicity evaluation, cells were incubated with Au NRs or Au NPs at the same gold concentration (40 µM, 80 µM, and 160 µM, respectively) for 24 h, and the cell viability was determined using the MTT (Sigma-Aldrich Co.) assay. Briefly, cells after different treatments were incubated with MTT (0.5 mg/mL in cell culture medium) for 2 h. The formazan crystals were dissolved with dimethyl sulfoxide (DMSO; Sigma-Aldrich Co.). Absorbance of the extracted solubilized formazan was measured at 570 nm by subtracting the background signal in the 96-well plate with a multimode microplate reader (Synergy HT; BioTek Instruments, Inc.).
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