Mcf 7

Manufactured by Keygen Biotech

Sourced in China

The MCF-7 is a well-established cell line derived from human breast adenocarcinoma. It is a commonly used model for the study of breast cancer biology and drug development.

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MCF-7/ADR (8 citations) MCF-7 cells (7 citations) MCF-7/DOX (4 citations) MCF-7/ADR cells (3 citations) MCF-7/DDP (2 citations)

37 protocols using mcf 7

Silencing UBE3C expression in breast cancer cells

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The MCF-10A, MCF-7, and MDA-MB-453 cell lines were obtained from KeyGEN BioTECH Inc. (Nanjing, China). MCF-7 and MDA-MB-453 cells were maintained in RPMI-1640 medium (KeyGEN BioTECH Inc.) supplemented with 10% (v/v) fetal bovine serum (FBS) at 37 °C with 5% CO₂. MCF-10A cells were cultured in DMEM/F12 (KeyGEN BioTECH Inc.) supplemented with 5% (v/v) horse serum, 20 ng/mL human EGF, 10 μg/mL insulin, 0.5 μg/mL hydrocortisone, penicillin, streptomycin and 100 ng/mL cholera toxin.
For subsequent assays, MCF-7 and MDA-MB-453 cells were transfected with UBE3C-siRNAs (The sequences of siRNAs for UBE3C were shown in Table 1), siRNA#NC, a UBE3C plasmid (Cloning vector: pcDNA3.1(+)-EGFP), or a control plasmid, which were synthesized by KeyGEN BioTECH Inc. (Nanjing, China), using Lipofectamine 2000 Reagent (Invitrogen) according to the manufacturer’s instructions.Table 1

The sequences of siRNAs for UBE3C

siRNAs	Sequences
siRNA#1	5′-UGAAGAAGCUGGACACAAATT-3’
siRNA#1	5′-UUUGUGUCCAGCUUCUUCATT-3’
siRNA#2	5′-GGAAGAAAGAAGAAAGAGATT-3’
siRNA#2	5′-UCUCUUUCUUCUUUCUUCCTT-3’
siRNA#3	5′-CCAUAGAAGUUGUAGGUCATT-3’
siRNA#3	5′-UGACCUACAACUUCUAUGGTT-3’
siRNA#NC	5′-UUCUCCGAACGUGUCACGUTT-3’
siRNA#NC	5′-ACGUGACACGUUCGGAGAATT-3’

Hang C., Zhao S., Wang T, & Zhang Y. (2021). Oncogenic UBE3C promotes breast cancer progression by activating Wnt/β-catenin signaling. Cancer Cell International, 21, 25.

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Establishing Doxorubicin-Resistant Breast Cancer Cell Line

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The human breast cancer cell line MCF-7 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The MCF-7/DOX-resistant cell line was established from parental MCF-7 and supplied by KeyGen Biotech. Co., Ltd (Nanjing, People’s Republic of China). The drug-resistant index of the MCF-7/DOX-resistant cell line was 54. Culture plates and dishes were purchased from Corning Incorporated (Corning, NY, USA). MCF-7 cells were cultured in Roswell Park Memorial Institute 1640 medium without folic acid, supplemented with 10% fetal bovine serum, 100 IU/mL penicillin and 100 µg/mL streptomycin sulfate. MCF-7/ADR cells were cultured in the same medium with the addition of 1,000 ng/mL of DOX. All the cells were cultured at 37°C in a humidifier with 5% CO₂ atmosphere. All the experiments were performed on the cells in the logarithmic phase of growth.

Hong W., Shi H., Qiao M., Gao X., Yang J., Tian C., Zhang D., Niu S, & Liu M. (2017). Rational design of multifunctional micelles against doxorubicin-sensitive and doxorubicin-resistant MCF-7 human breast cancer cells. International Journal of Nanomedicine, 12, 989-1007.

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Breast Cancer Cell Line Cultivation and Manipulation

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Human breast cancer cell lines (MDA-MB-231 and MCF-7) were obtained from the KeyGEN BioTECH (Nanjing, China). MDA-MB-231 and MCF-7 cells were cultured in RPMI-1640 and Leibovitz's L-15 medium (KeyGEN BioTECH, Nanjing, China) supplied with 5% CO₂, respectively. Media in all cases were supplemented with 10% fetal bovine serum (Hyclone, Logan, UT), 100 units/mL penicillin, and 100 μg/mL streptomycin. For subsequent functional assays, MCF-7 and MDA-MB-231 cells were transfected with GNG7-siRNA, GNG7 plasmid using Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, USA) according to the manufacturer's instructions or exposed to OST.

Mei J., Wang T., Zhao S, & Zhang Y. (2021). Osthole Inhibits Breast Cancer Progression through Upregulating Tumor Suppressor GNG7. Journal of Oncology, 2021, 6610511.

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BCa Cell Lines Transfection Assay

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Human BCa cell lines MCF-7 and MDA-MB-231 were purchased from KeyGEN BioTECH Inc. (Nanjing, China). MCF-7 cells were cultured in RPMI-1640 media (KeyGEN BioTECH Inc.) containing 10% fetal bovine serum (FBS). MDA-MB-231 cells were cultured in L15 media (KeyGEN BioTECH Inc.) containing 10% FBS. All cells were cultured in the condition of 37 °C with 5% CO₂. For subsequent assays, COL10A1-siRNA and siRNA control, synthesized in KeyGEN BioTECH Inc. (Nanjing, China), using Lipofectamine 3000 Reagent (Invitrogen) according to the manufacturer's instructions, was used to transfect incubated cells.

Zhou W., Li Y., Gu D., Xu J., Wang R., Wang H, & Liu C. (2022). High expression COL10A1 promotes breast cancer progression and predicts poor prognosis. Heliyon, 8(10), e11083.

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Cell Line Establishment Protocol

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Cell lines used in this study, which included lines derived from hepatocellular carcinoma cells (HpG2), lung cancer cells (NCI-A549, hereafter referred to as A549 cells; NCI-H1299, hereafter referred to as H1299 cells), breast cancer cells (MCF-7), colorectal cancer cells (HCT-8), and pancreatic cancer cells (PANC-1), were obtained from KeyGen Biotech (Nanjing, China). All cells were cultured in RPMI 1640 or DMEM media supplemented with 10% fetal bovine serum under conditions of 37°C, 95% humidity, and 5% CO₂ (hereafter referred to as standard cell culture conditions). No primary human tumor specimens were used in this study.

Yang L., Jin W.Q., Tang X.L., Zhang S., Ma R., Zhao D.Q, & Sun L.W. (2022). Ginseng-derived nanoparticles inhibit lung cancer cell epithelial mesenchymal transition by repressing pentose phosphate pathway activity. Frontiers in Oncology, 12, 942020.

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Extraction and Analysis of Total RNA from HEK-293 and MCF-7 Cells

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Human embryonic kidney cells HEK-293 and human breast cancer cells MCF-7 (KeyGEN Biotech, Nanjing, China) were grown in a clean incubator at 37 C with a CO 2 content of 5%.
The medium was a DMEM high-sugar medium containing 100 U ml À1 penicillin, 100 mg ml À1 streptomycin (KeyGEN Biotech, Nanjing, China) and 10% fetal bovine serum (FBS, Gibco, Scotland, UK). Total RNA was extracted by the Total RNA Extraction Kit (Omega, Norcross, USA). Before the total RNA extraction, the medium in the culture dish was aspirated, and the TRK lysis buffer containing mercaptoethanol was directly added to the culture dish to obtain a cell lysate. In the immediate aermath of the column of the Total RNA Extraction Kit was centrifuged and washed several times, the total RNA extracts of the two cells were obtained aer appropriate dilution in DEPC water. NanoDrop 2000 was used to detect the concentration, and the extras were immediately put in a À80 C refrigerator aer indicating the label.

Zhu Q., Li H., & Xu D. (2020). Sensitive and enzyme-free fluorescence polarization detection for miRNA-21 based on decahedral sliver nanoparticles and strand displacement reaction. RSC Advances, 10(29), 17037-17044.

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Ovarian and Breast Cancer Cell Lines

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The human ovarian cancer cell line A2780 and its related cDDP-resistant cell lines (ACRP) were obtained from NANJING KEYGEN BIOTECH CO., LTD, China. SKOV3, OVCAR3, OV2008 and C13* cells, a human breast cancer cell line (MCF-7) and the ADM-resistant cell line (MCF-7/ADM) were obtained from the Shanghai Institute of Cell Biology at the China Academy of Sciences. The culture conditions are described in the Supplementary Methods.

Zhu X., Shen H., Yin X., Long L., Chen X., Feng F., Liu Y., Zhao P., Xu Y., Li M., Xu W, & Li Y. (2017). IL-6R/STAT3/miR-204 feedback loop contributes to cisplatin resistance of epithelial ovarian cancer cells. Oncotarget, 8(24), 39154-39166.

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Establishing Paclitaxel-Resistant Breast Cancer Cells

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Breast cancer cells (MCF‐7 and MDA‐MB‐468) and normal breast epithelial cells (MCF‐10A) were acquired from Nanjing KeyGen Biotech (China). Breast cancer cells were treated with increasing concentrations of PTX to establish PTX‐resistant cells (MCF‐7/PTX and MDA‐MB‐468/PTX). In brief, MCF‐7 and MDA‐MB‐468 cells were seeded in 96‐well plates and the detection of the half‐maximal inhibitory concentration (IC₅₀ value) of PTX was performed. Cells in six‐well plates were exposed to PTX at a 1/50 concentration of IC₅₀ until their stable growth. Subsequently, the concentration of PTC was increased in multiples until the end of the fifth month, with 2‐week culture for each concentration. These cells were maintained in Dulbecco's modified Eagle medium (DMEM; HyClone) containing 10% (v/v) fetal bovine serum (FBS; Sigma) at 37°C with 5% CO₂.

Kong Z., Han Q., Zhu B., Wan L, & Feng E. (2023). Circ_0069094 regulates malignant phenotype and paclitaxel resistance in breast cancer cells via targeting the miR‐136‐5p/YWHAZ axis. Thoracic Cancer, 14(19), 1831-1842.

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Culturing Breast Cancer Cell Lines

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The human breast cancer cell lines MCF-7 and MCF-7/Dox cells were purchased from KeyGen Biotech Co., Ltd.(Nanjing China) and cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) and 100 units/mL penicillin, 100 mg/mL streptomycin at 37°C and 5% CO₂. MCF-7/Dox cells were cultured in medium with 2 mg/mL doxorubicin and grown in doxorubicin-free culture medium for more than 2 weeks before assay.

Xue C., Wang C., Sun Y., Meng Q., Liu Z., Huo X., Sun P., Sun H., Ma X., Ma X., Peng J, & Liu K. (2016). Targeting P-glycoprotein function, p53 and energy metabolism: Combination of metformin and 2-deoxyglucose reverses the multidrug resistance of MCF-7/Dox cells to doxorubicin. Oncotarget, 8(5), 8622-8632.

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Cell Culture Protocols for Cancer Research

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The human carcinoma cell lines A549, HEK-293, PLC-PRF-5, H446, LLC, MCF-7, MDA-MB-231, HCT-116 and BL16F10 were obtained from KeyGen Biotech (Nanjing, China). The cells were cultured in RPMI-1640 medium (Hyclone, USA) supplemented with 10% fetal bovine serum (Hyclone, USA) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin). All cultures were maintained at 37°C in a humidified atmosphere containing 5% CO₂.

Zhong W., Chen S., Zhang Q., Xiao T., Qin Y., Gu J., Sun B., Liu Y., Jing X., Hu X., Zhang P., Zhou H., Sun T, & Yang C. (2017). Doxycycline directly targets PAR1 to suppress tumor progression. Oncotarget, 8(10), 16829-16842.

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