The human SCLC cell line NCI-H69 and the drug-resistant sublines NCI-H69AR, NCI-H446, NCI-H146, and NCI-H1688 were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA) and maintained in RPMI 1640 medium (HyClone, Logan, UT, USA) with 10 % fetal calf serum (HyClone) in an incubator at 37 °C with 5 % CO2. The H69AR subline was maintained in a 5 μg/L final concentration of doxorubicin (Jiangshu, China) and transferred to drug-free media for at least 2 weeks before any experiment.
Nci h69ar
Manufactured by American Type Culture Collection
Sourced in United States
The NCI-H69AR is a cell line derived from a human small cell lung carcinoma. It is resistant to the chemotherapeutic agent adriamycin.
Automatically generated - may contain errors
Lab products found in correlation
Nci h69, by American Type Culture Collection (10 mentions)
Nci h446, by American Type Culture Collection (9 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Nci h1688, by American Type Culture Collection (1 mentions)
Nci h146, by American Type Culture Collection (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Tapestation 4200, by Agilent Technologies (1 mentions)
Novaseq 6000 platform, by Illumina (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Penicillin, by Beyotime (1 mentions)
Streptomycin, by Beyotime (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Rpmi 1640 medium, by Cytiva (1 mentions)
Opti mem 1, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
10 protocols using nci h69ar
1
Culturing Small Cell Lung Cancer Cell Lines
2
Establishing Chemoresistant SCLC Cell Lines
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Human SCLC cell lines (NCI‐H69, NCI‐H69AR, and NCI‐H446) were obtained from the American Type Culture Collection (Manassas, VA, USA). The chemoresistant subline H446CDDP was induced by culturing H446 cells in progressively increasing concentrations of cisplatin (up to 0.5 μg/ml) over 12 months. H446CDDP was cultured in RPMI 1640 medium (Gibco, Waltham, WA, USA) supplemented with 10% FBS (Gibco, Waltham, WA, USA) and cisplatin (0.5 μg/ml). H69 and H446 cells were cultured in RPMI 1640 medium supplemented with 10% FBS, while H69AR was cultured in RPMI 1640 medium supplemented with 20% FBS. The chemoresistant sublines H69AR and H446CDDP were transferred to a drug‐free medium for at least 2 weeks before any experiment. All cell lines were maintained in a humidified incubator at 37°C with the presence of 5% CO2. All cell lines were not contaminated with mycoplasma.
3
Establishing SCLC Cell Line Cultures
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The human SCLC cell lines NCI-H69 and NCI-H446, and the chemoresistant cell line NCI-H69AR were purchased from the American Type Culture Collection (USA). The CDDP-resistant NCI-H446DDP cell line was constructed in our laboratory (14 (link)). These cell lines were all cultured in RPMI-1640 (Invitrogen) supplemented with 10% fetal bovine serum (HyClone) at 37 ℃ with 5% carbon dioxide.
4
Small Cell Lung Cancer Cell Lines
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The human SCLC cell lines NCI-H69, NCI-H69AR, NCI-H446, and 16-HBE cells were all from the American Type Culture Collection (ATCC; Manassas, VA, USA), maintained at 37°C in a 5% CO2 humidified incubator and grown in Roswell Park Memorial Institute (RPMI)-1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS (HyClone, Logan, UT, USA). The H69AR cell line is a multidrug-resistant cell line that is primarily used in microarrays to screen for chemoresistance-related miRNAs in SCLC and is occasionally used in this study.7 (link)
5
Establishing SCLC cell lines and drug-resistant sublines
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Human SCLC cell line NCI-H69, NCI-H446 and the drug-resistant subline NCI-H69AR were purchased from the American Type Culture Collection (ATCC, USA) and maintained in RPMI 1640 medium contain in L-Glutamine with 10% and 20% fetal calf serum respectively in an incubator at 37°C with 5% CO2. The adriamycin-resistant NCI-H446 cell line (NCI-H446AR) was obtained by culturing these cells in gradually increasing doses of adriamycin up to 0.8 uM after a total of 14 months in our laboratory. The drug-resistant cells were maintained in drug-free medium for at least 2 weeks before any experiment.
6
Establishing drug-resistant SCLC cell lines
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The human SCLC cell lines, NCI-H69 and NCI-H446, and the multidrug-resistant cell line, NCI-H69AR, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), as described in our previous study (17 (link)). A drug-resistant subline, H446DDP, was established in our laboratory by culturing H446 cells in cisplatin (DDP; Shandong, China). Cells were maintained in RPMI-1640 medium containing 10% fetal bovine serum (FBS; Gibco, CA, USA) at 37 °C in a humidified atmosphere containing 5% CO2.
7
Maintaining NCI-H69 and NCI-H69AR Cell Lines
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Human SCLC cell line NCI-H69 and the drug-resistant subline NCI-H69AR were purchased from the American Type Culture Collection (ATCC, USA) and maintained in RPMI 1640 medium containing L-glutamine with 10% and 20% foetal calf serum, respectively, in an incubator at 37°C with 5% CO 2 . The drug-resistant variant H69AR was alternately treated with drug-free medium and medium containing 5 μg/ml of adriamycin (ADM) and tested regularly for maintained resistance to the selected drugs by CCK-8 assay.
8
Transcriptomic Profiling of SCLC Cell Lines
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Human SCLC cell lines, namely, NCI-H69, NCI-H69AR, and NCI-H446, were acquired from the American Type Culture Collection (ATCC, USA). Chemo-resistant H446DDP cells were obtained by incubating H446 cells in progressively increasing cisplatin doses of up to 5 μg/mL for 6 months. RNA was extracted from human cell lines such as NCI-H69, NCI-H69AR, NCI-H446, and NCI-H446DDP by using Trizol method and quantified by Qubit3.0. RNA quality was assessed by 4200 TapeStation (Agilent Technologies). RNA libraries were prepared with VAHTSTMmRNA-seq V3 preparation kit (Illumine). The libraries were then quantified and sequenced with Illumina Novaseq6000 platform.
9
Establishment of Cisplatin-Resistant SCLC Cell Lines
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Human SCLC cells lines NCI-H69, NCI-H446 and NCI-H69AR were purchased from American Type Culture Collection. Cisplatin (DDP)-resistant NCI-H446DDP cells were established by exposing H446 cells to DDP, as previously described (16 (link)). All cells were cultured in RPMI-1640 medium (Cytiva) containing 10% fetal calf serum (Gibco; Thermo Fisher Scientific, Inc.) and 1% antibiotics (100 mg/ml penicillin and 100 mg/ml streptomycin; Beyotime Institute of Biotechnology) in an incubator at 37˚C with 5% CO2. The following antibodies were used: Anti-fibroblast growth factor receptor-like 1 (FGFRL1; cat. no. ab95940) and anti-HuR (cat. no. ab200342) (both from Abcam); and anti-GAPDH (cat. no. BS72410; Bioworld Technology, Inc.).
10
Generation of Chemoresistant SCLC Cell Lines
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The human SCLC cell lines NCI-H69, NCI-H69AR and NCI-H446 were acquired from the American Type Culture Collection (ATCC, USA). Chemoresistant H446DDP cells were acquired by incubating H446 cells in progressively increasing doses of cisplatin (up to 5 μg/mL) over a period of 6 months. Cells were transiently transfected with siRNAs for CDKN1C (GenePharma, Shanghai, China) by using Lipofectamine 2000 and OPTI-MEM I (Invitrogen, USA) according to the manufacturer's instructions. For stable expression, lentiviral particles expressing shRNA for CDYL (shCDYL#1 and shCDYL#2), LV5-CDYL, and pcDNA3.1-CDKN1C (GenePharma, Shanghai, China) were transfected into SCLC cells. The sequences of shRNA and siRNA are listed in Table S1 -2.
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