Murine colon carcinoma CT26, human bladder carcinoma J82, T24, SW1710, and UM-UC-3, breast cancer MCF-7 and T47D, colorectal cancer DLD-1, HCT15, HT29, Lovo, and HCT116, hepatoma HepG2, non-small cell lung cancer NCI-H460, ovary cancer Caov-3, prostate cancer LNCap clone FGC, and renal carcinoma Caki-1, as well as normal HUVEC, intestinal epithelial FHs74Int, and lung fibroblast MRC-5 cell lines were obtained from the National Collection of Authenticated Cell Cultures. Murine colorectal adenocarcinoma MC38 cell lines were purchased from MingzhouBio, Ningbo, Zhejiang, China. The cells were maintained in the logarithmic phase of growth in DMEM, MEM, McCoy’5a, or RPMI medium supplemented with 10% heat-inactivated FBS, 100 IU/mL penicillin, and 100 μg/mL streptomycin in an incubator at 37 °C with a humidified atmosphere of 5% CO2. Cell line identity was validated through short tandem repeat profiling, and routine mycoplasma testing was negative for contamination.
Nci h460
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China
The NCI-H460 is a cell line derived from a human large cell lung carcinoma. It is a commonly used model for studying lung cancer biology and evaluating potential therapeutic agents.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (18 mentions)
Nci h1299, by National Collection of Authenticated Cell Cultures (13 mentions)
Nci h1975, by National Collection of Authenticated Cell Cultures (6 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Hepg2, by National Collection of Authenticated Cell Cultures (4 mentions)
Hcc827, by National Collection of Authenticated Cell Cultures (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (4 mentions)
Beas 2b, by National Collection of Authenticated Cell Cultures (4 mentions)
Dld 1, by National Collection of Authenticated Cell Cultures (3 mentions)
Hct116, by National Collection of Authenticated Cell Cultures (3 mentions)
Mrc 5, by National Collection of Authenticated Cell Cultures (3 mentions)
Nci h292, by National Collection of Authenticated Cell Cultures (3 mentions)
Nci h1650, by National Collection of Authenticated Cell Cultures (3 mentions)
Spc a1, by National Collection of Authenticated Cell Cultures (3 mentions)
Nci h460, by Thermo Fisher Scientific (3 mentions)
Packaging kit, by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Sw1710, by National Collection of Authenticated Cell Cultures (2 mentions)
41 protocols using nci h460
1
Cell Line Maintenance and Characterization
2
Cell Line Maintenance and Characterization
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Murine colon carcinoma CT26, human bladder carcinoma J82, T24, SW1710, and UM-UC-3, breast cancer MCF-7 and T47D, colorectal cancer DLD-1, HCT15, HT29, Lovo, and HCT116, hepatoma HepG2, non-small cell lung cancer NCI-H460, ovary cancer Caov-3, prostate cancer LNCap clone FGC, and renal carcinoma Caki-1, as well as normal HUVEC, intestinal epithelial FHs74Int, and lung fibroblast MRC-5 cell lines were obtained from the National Collection of Authenticated Cell Cultures. Murine colorectal adenocarcinoma MC38 cell lines were purchased from MingzhouBio, Ningbo, Zhejiang, China. The cells were maintained in the logarithmic phase of growth in DMEM, MEM, McCoy’5a, or RPMI medium supplemented with 10% heat-inactivated FBS, 100 IU/mL penicillin, and 100 μg/mL streptomycin in an incubator at 37 °C with a humidified atmosphere of 5% CO2. Cell line identity was validated through short tandem repeat profiling, and routine mycoplasma testing was negative for contamination.
3
Cell Growth Inhibition Assay of Peptides
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The HepG2, A549, and NCI-H460 cell lines were purchased from the National Collection of Authenticated Cell Cultures (NCACC). HepG2 (Serial No. SCSP-510) and A549 (Serial No. SCSP-503) were maintained in DMEM media containing 10% FBS, while NCI-H460 (Serial No. SCSP-584) was cultured in RPMI-1640 media plus 10% FBS at 37 °C with 5% CO2.
For the cell growth inhibition assay, 4 × 103 cells in 100 µL of medium were seeded per well in 96-well plates. Twelve hours later, wells in the control group were replaced with fresh medium, while those in the treatment group were supplemented with fresh medium containing peptides at the indicated concentration. After another incubation for 48 h, the cell growth inhibition rate was measured using the CCK-8 assay, following the instructions from the user’s manual. The inhibition rate was calculated as follows:
Inhibition rate = [1 − (At − Ab)/(Ac − Ab)] × 100%, where At = absorbance value of the treatment group, Ac = absorbance value of the control group, and Ab = absorbance value of the blank respectively. The IC50 value was calculated by the modified Karber method.
For the cell growth inhibition assay, 4 × 103 cells in 100 µL of medium were seeded per well in 96-well plates. Twelve hours later, wells in the control group were replaced with fresh medium, while those in the treatment group were supplemented with fresh medium containing peptides at the indicated concentration. After another incubation for 48 h, the cell growth inhibition rate was measured using the CCK-8 assay, following the instructions from the user’s manual. The inhibition rate was calculated as follows:
Inhibition rate = [1 − (At − Ab)/(Ac − Ab)] × 100%, where At = absorbance value of the treatment group, Ac = absorbance value of the control group, and Ab = absorbance value of the blank respectively. The IC50 value was calculated by the modified Karber method.
4
MTT Cytotoxicity Assay of Cancer Cell Lines
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Human lung cancer NCI-H460, human liver cancer HePG-2, human breast carcinoma MCF-7 and MDA-MB-231 cell lines were obtained from the National Collection of Authenticated Cell Cultures (Shanghai, China). All cells were incubated at 37°C in a humidified incubator with a 5% CO2 incubator. Cells after the third generation were used for further experiment.
The assay of 3-(4,5-dimethylthiazole-2-yl)-2,5-dip-henyltetrazolium bromide (MTT) was carried out against the MDA-MB-231, HePG-2, NCI-H460, and MCF7 cell lines. Test samples were dissolved in dimethyl sulfoxide (DMSO) to final concentrations of 200 μM in 96-well plates and each well was placed with 200 μL of cells (3 × 103 per well for cancer cell lines). All the treatments were replicated in triplicate and an equivalent volume of DMSO was used as blank control.
The assay of 3-(4,5-dimethylthiazole-2-yl)-2,5-dip-henyltetrazolium bromide (MTT) was carried out against the MDA-MB-231, HePG-2, NCI-H460, and MCF7 cell lines. Test samples were dissolved in dimethyl sulfoxide (DMSO) to final concentrations of 200 μM in 96-well plates and each well was placed with 200 μL of cells (3 × 103 per well for cancer cell lines). All the treatments were replicated in triplicate and an equivalent volume of DMSO was used as blank control.
5
Cultivation and Maintenance of Common Cell Lines
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Human lung cancer NCI-H460, human breast carcinoma MCF-7 and MDA-MB-231 cell lines were obtained from the National Collection of Authenticated Cell Cultures (Shanghai, China). All cells were incubated at 37°C in a humidified incubator with 5% CO2 incubator. Cells after the third generation were used for experiment.
6
Human NSCLC Cell Line Culture
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Human NSCLC cell lines A549, 95C, 95D, HBE, NCI-H1299, and NCI-H460 were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). These cells were routinely cultured in DMEM or RPMI-1640 medium (HyClone, USA) (HyClone, Logan City, USA) supplemented with 10% fetal bovine serum (Gibco, Carlsbad, USA) and 1% penicillin/streptomycin (100 IU/ml) at 37°C in a humidified incubator with 5% CO2.
7
Quantitative Analysis of ARTN Expression in NSCLC
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Human NSCLC cell lines (NCIH1299, PC9, NCIH460, NCIH292) and human bronchial epithelial cells (HBE) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). RNA was extracted from cell samples and patient samples using TRIzol RNA Extraction Reagent (Invitrogen™, 15596026) according to the instructions. After removal of residual gDNA, reverse transcription reactions were performed using 1ug of total RNA as template. This was followed by real-time quantitative PCR(qPCR) using 2 × qPCR MasterMix(KAPA, KM4103) on a LightCycler 480 instrument (Roche). Relative gene expression was analyzed by the 2-ΔΔCt method, using Gapdh as an internal control. qPCR and in vitro cancer cell function assays were also performed as described previously [33 (link), 34 (link)]. qPCR primer sequences are shown in Additional file 1 : Table S3. siRNA sequences of ARTN were purchased from GenePharma Co. The sequences are listed in Additional file 1 : Table S4. In addition, immunohistochemical assays were used to verify the difference in protein expression of ARTN in normal lung tissue and lung cancer tissue.
8
Establishing and Culturing Human Cancer Cell Lines
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Human ovarian cancer cell line HO-8910 and its derivative HO-8910PM with high metastatic capability were established in our previous studies [23 , 24 (link)]; human NSCLC cell lines A549, NCI-H460, NCI-H1975, NCI-H1395, U1752 and SK-MES-1 were purchased from the Cell Bank of the Chinese Academy of Sciences, Shanghai, China. SK-MES-1 cells were cultured in Minimum Essential Medium (MEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco); other cells were cultured in RPMI 1640 medium (Gibco) supplemented with 10% FBS. All cells were maintained in a 37°C, 5 % CO2 incubator.
9
Cell Culture Protocol for NSCLC and HEK-293T
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Human NSCLC cell lines (A549, HCC827, NCI-H1299, NCI-H1650, NCI-H226, and NCI-H460) and HEK-293T cells were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). These cells were routinely cultured in DMEM medium (HyClone, Logan City, USA) supplemented with 10% fetal bovine serum (Gibco, Carlsbad, USA) and 1% penicillin/streptomycin (100 IU/mL) at 37 °C in a humidified incubator with 5% CO2.
10
Lung Cancer Cell Lines: Cultivation and Demethylation
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Human lung carcinoma cell lines (A549, SPC-A-1, NCI-H460, and SK-MES-1) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). A549, SPC-A-1, and NCI-H460 cells were seeded and grown in RPMI 1640 medium (HyClone, South Logan, UT, USA) with 10% heat-inactivated fetal bovine serum (Gibco, Carlsbad, CA, USA). SK-MES-1 cells were cultured in MEM (Gibco) with 10% fetal bovine serum (Gibco), L-glutamine, and antibiotics (Invitrogen, Carlsbad, CA, USA). All cells were cultured in a humidified incubator containing 5% CO2 at a temperature of 37°C. 5-aza-2’-deoxycytidine (Sigma–Aldrich, St. Louis, MO, USA) was used as a demethylating agent to treat the cells. The drug treatment protocol was described previously by us [29 (link)].
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