Human normal lung epithelial cell line BEAS‐2B and LUAD cell line NCI‐H1975 were purchased from Fenghui Biotechnology Co., Ltd. LUAD cell line NCI‐H441 was purchased from BeNa Culture Collection (BNCC). BEAS‐2B cells were cultured in DMEM medium (Gibco, C11995500BT) with 1% penicillin (HyClone, SV30010) and 10% FBS (Gibco, 10099–141). NCI‐H441 and NCI‐H1975 cells were cultured in 1640 medium (Gibco, C11875500BT) with 1% penicillin (HyClone, SV30010) and 10% FBS (Gibco, 10099–141). The cells were cultured at 37°C in an incubator containing 5% CO2. Then, the total RNA was extracted with TRIZOL reagent (Tiangen, dp424, China) and detected for concentration and purity using ultramicro ultraviolet–visible spectrophotometer Nanodrop 2000 (Thermo). After reverse transcription (RevertAid First Strand cDNA Synthesis Kit; Thermo), PCR was performed with a fluorescent quantitative PCR detector (ABI Veriti; Life Technologies) using TB Green Premix Ex Taq II (Takara, RR820A). GAPDH was used as a reference gene. Three repeat samples were taken for each sample (see Table 2 for primer sequences). The mRNA expression level was calculated by 2−△△CT.
Nci h441
Manufactured by Thermo Fisher Scientific
Sourced in United States, Switzerland
The NCI‐H441 is a human lung adenocarcinoma cell line. It is derived from a primary lung adenocarcinoma tumor. The NCI‐H441 cell line is commonly used in cell-based research and assays.
Automatically generated - may contain errors
Lab products found in correlation
Beas 2b, by Thermo Fisher Scientific (2 mentions)
Nanodrop 2000, by Thermo Fisher Scientific (2 mentions)
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions)
Nci h1975, by Thermo Fisher Scientific (1 mentions)
C11875500bt, by Thermo Fisher Scientific (1 mentions)
Abi veriti, by Thermo Fisher Scientific (1 mentions)
Sv30010, by Thermo Fisher Scientific (1 mentions)
Tb green premix ex taq 2, by Takara Bio (1 mentions)
Trizol reagent, by Tiangen Biotech (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Sodium bicarbonate, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Merck Group (1 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
Mrc 5 cells, by Thermo Fisher Scientific (1 mentions)
24 well plate, by Corning (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hcc827, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Nunc delta surface, by Thermo Fisher Scientific (1 mentions)
4 protocols using nci h441
1
Profiling LUAD cell gene expression
2
Culturing MRC-5 and NCI-H441 Cell Lines
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MRC-5 cells (healthy human lung fibroblast cell line) and NCI-H441 (alveolar epithelial cell line) were purchased from the American Type Cell Culture Collection (ATCC, Manassas, VA, USA). MRC-5 cells were maintained in modified eagle medium (MEM; Gibco), supplemented with 10% (v/v) foetal bovine serum (FBS; Invitrogen), 1% (v/v) sodium pyruvate (Gibco), 7.5% (v/v) sodium bicarbonate (Gibco) and 1% (v/v) non-essential amino acids (Sigma-Aldrich, Australia) then incubated at 37 °C with CO2. MRC-5 cells were seeded in 24-well plates (Corning Costar, Cambridge, MA, USA) at a density of 6 × 104 cells/cm2.
NCI-H441 cells were maintained in RPMI 1640 media (Gibco) supplemented with 10% (v/v) FBS (Invitrogen) at 37 °C with 5% CO2 (v/v). NCI-H441 cells were seeded at a density of 3 × 105/cm2. All experiments were conducted between a population doubling level of 28–42 for the MRC-5 cell line and between passage numbers 45–78 for the NCI-H441 cell line.
NCI-H441 cells were maintained in RPMI 1640 media (Gibco) supplemented with 10% (v/v) FBS (Invitrogen) at 37 °C with 5% CO2 (v/v). NCI-H441 cells were seeded at a density of 3 × 105/cm2. All experiments were conducted between a population doubling level of 28–42 for the MRC-5 cell line and between passage numbers 45–78 for the NCI-H441 cell line.
3
Surgical LUAD Tissue Collection Protocol
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This study was approved by the Ethics Committees of Liaoning Cancer Hospital and Institute, and conducted in accordance with the Declaration of Helsinki. All patients provided written informed consent before inclusion. Patients were selected from LUAD patients receiving surgical treatment at Liaoning Cancer Hospital and Institute from April 2012 to October 2015. The patients who received any type of therapy for cancer before surgery were excluded. Based on this, 105 paired tissues with adequate clinical and histological characteristics were collected in the study according to the criteria proposed by the IASLC/ATS/ERS in 2011.17 (link) TNM stage was determined according to the 7th edition of the American Joint Committee on Cancer (AJCC) cancer staging manual.
Four kinds of human LUAD cells, A549, HCC827, NCI-H441, and NCI-H1734, and an immortalized human bronchial epithelial cell line (BEAS-2B) were obtained from ATCC (Rockville, MD, USA) and cultured in RPMI medium 1640 (Invitrogen, USA) supplemented with 10% FBS (Invitrogen, USA). All cells were maintained at the condition of 5% CO2 and 37°C.
Four kinds of human LUAD cells, A549, HCC827, NCI-H441, and NCI-H1734, and an immortalized human bronchial epithelial cell line (BEAS-2B) were obtained from ATCC (Rockville, MD, USA) and cultured in RPMI medium 1640 (Invitrogen, USA) supplemented with 10% FBS (Invitrogen, USA). All cells were maintained at the condition of 5% CO2 and 37°C.
4
Comparative analysis of immortalized lung cell lines
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Immortalized lung cell lines, Calu-3, BEAS2-B, NCI-H292, NCI-H441, and A549, were obtained from the American Type Culture Collection (ATCC) (Manassas, Virginia) in Nippon Boehringer Ingelheim and used for this study. These cell lines were seeded at a density of 1 × 10 5 cells/cm 2 on the 75 cm 2 of Nunc Delta Surface (Lot no. 12989; Nunc, Roslilde, Denmark). Calu-3 (passage number: 19-21) was cultured in EMEM (Lonza, Basel, Switzerland), BEAS2-B (passage number: 41-50) in BEGM (Lonza), NCI-H292 (passage number: 10-12), and NCI-H441 (passage number: 12-13) in RPMI1640 (Invitrogen, Carlsbad, California), and A549 (passage number: 80-90) in DMEM (Lonza), including 10% fetal bovine serum (Moregate, Bulimba, Australia), for 7 days at 37°C in 5% CO 2 , and the medium was exchanged every 2 days. All peptides were selected according to the in silico selection criteria reported previously 14 and synthesized by Thermoelectron Corporation (Sedanstrabe, Germany) at a peptide purity of more than 95%. The concentrations of peptide solutions were determined by quantitative amino acid analysis (Lachrom Elite, Hitachi, Tokyo, Japan). Other chemicals used were commercial products of an analytical grade.
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