Nci h1299

Manufactured by National Collection of Authenticated Cell Cultures

Sourced in China

NCI-H1299 is a human non-small cell lung cancer cell line. It is a widely used model for cancer research, particularly in the study of lung cancer biology and the evaluation of therapeutic interventions.

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H1299 (168 citations)

50 protocols using nci h1299

NSCLC Tumor Sample Collection and Cell Line Cultivation

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Ninety-six tumor samples and their adjacent normal tissues (2 cm from the lesion) were obtained from patients with primary NSCLC in the Sichuan Cancer Hospital & Institute, Sichuan Cancer Center, between March 2014 and September 2016. None of the patients had received radiotherapy or chemotherapy before the surgical operation. The study was approved by the Ethics Committee of Sichuan Cancer Hospital & Institute, Sichuan Cancer Center and was conducted following the Declaration of Helsinki (2013 revision). The patients provided informed written consent. Samples were kept in liquid nitrogen for analysis.
The human NSCLC cell lines A549, NCI-H1299, HCC827, NCI-1650, and NCI-H358 were acquired from the National Collection of Authenticated Cell Cultures (Shanghai, China), and the normal human lung epithelial cell line DEAS-2B was obtained from Beyotime (Shanghai, China). A549 cells were maintained in DMEM/F12 medium (Procell, Wuhan, China) with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA). The remaining cell lines were cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 1% Glutamax (Invitrogen), 1% (v/v) 100 mM sodium pyruvate solution (Invitrogen, Waltham, MA, USA), and 10% FBS. All cells were grown in a humidified 37 °C incubator with 5% CO₂.

Zhang Y., Yuan J., Guo M., Xiang R., Xie T., Zhuang X., Dai W., Li Q, & Lai Q. (2023). Upregulation of long intergenic non-coding RNA LINC00326 inhibits non-small cell lung carcinoma progression by blocking Wnt/β-catenin pathway through modulating the miR-657/dickkopf WNT signaling pathway inhibitor 2 axis. Biology Direct, 18, 3.

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Investigating MK-2206 inhibition in LUAD cell lines

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Two human LUAD cell lines, A549 and NCI-H1299, were obtained from the National Collection of Authenticated Cell Cultures (Shanghai, China). Cells were cultured in RPMI-1640 medium (Gibco, Waltham, MA, USA) with 10% FBS (BI, Tel Aviv, Israel) at 37 °C and 5% CO₂. MK-2206 (Selleck, Shanghai, China), inhibiting the phosphorylation process against Akt1, Akt2 and Akt3, was diluted with DMSO (Sigma, Waltham, MA, USA) to 20 mM. Next, the solution was diluted to 1 mM with RPMI-1640 and stored at −20 °C. Cells were treated with MK-2206 at 5 µM after culturing for 24 h, which was marked as the 0 h in MK-2206 group.

Zhang N., Cao S., Sun R., Wang Y., Liu L., Wang W, & Meng X. (2022). Signal peptidase 21 suppresses cell proliferation, migration, and invasion via the PTEN-PI3K/Akt signaling pathway in lung adenocarcinoma. PeerJ, 10, e14206.

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Knockdown of Keratins in Lung Cancer Cells

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Two human lung cancer cells NCI–H1299 and A549, were purchased from National Collection of Authenticated Cell Cultures (Shanghai, China). These cells were cultured in DMEM (Gibco, Carlsbad, CA, United States) supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, United States) and 1% penicillin–streptomycin at 37 °C in a 5% CO₂ humidified atmosphere incubator. The small interfering RNAs (siRNA) designed to target KRT7 (si-KRT7) and KRT8 (si-KRT8), along with non-specific control siRNA (si-Control) were synthesized by RiboBio (Guangzhou, China). Transfection of si-KRT7, si-KRT8 and si-Control into NCI–H1299 and A549 cells was performed utilizing Lipofectamine RNAi Max (Thermo Fisher, Waltham, MA, United States) following the manufacturer’s instructions. ns, not significant, **P < 0.05, **P < 0.01 and ***P < 0.001.

Li G., Guo J., Mou Y., Luo Q., Wang X., Xue W., Hou T., Zeng T, & Yang Y. (2024). Keratin gene signature expression drives epithelial-mesenchymal transition through enhanced TGF-β signaling pathway activation and correlates with adverse prognosis in lung adenocarcinoma. Heliyon, 10(3), e24549.

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Evaluation of Epigenetic Drugs in Cell Lines

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HEK293 (human embryonic kidney 293 cells), A549, NCI-H1299, and NCI-H520 (human lung cancer cells) cells were purchased from National Collection of Authenticated Cell Cultures. IMR90 cells were purchased from the American Type Culture Collection (ATCC). IMR90 cells were cultured in ATCC-formulated Eagle’s minimum essential medium supplemented with 10% fetal bovine serum (FBS; Gibco). HEK293 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Invitrogen, 11960044) supplemented with 10% FBS (Gibco), 1% GlutaMAX (Invitrogen, 35050061), 1% sodium pyruvate (Invitrogen, 11360070), streptomycin (100 μg/ml), and penicillin (100 U/ml). A549 cells were cultured in F-12 K (Invitrogen, 21127-022) supplemented with 10% FBS, streptomycin (100 μg/ml), and penicillin (100 U/ml). NCI-H1299 cells were cultured in RPMI 1640 (Invitrogen, 11875093) supplemented with 10% FBS, 1% GlutaMAX, 1% sodium pyruvate, streptomycin (100 μg/ml), and penicillin (100 U/ml). NCI-H520 cells were cultured in RPMI 1640 supplemented with 10% FBS, streptomycin (100 μg/ml), and penicillin (100 U/ml). All cell lines were maintained at 37°C in a humidified incubator with 5% CO₂. NCI-H1299, IMR90, and HEK293 were treated with 500 nM DAC, 500 nM SB939, or 500 nM DAC + 500 nM SB939 for 72, 18, or 72 + 18 hours, respectively, and compound-containing medium was refreshed every 24 hours.

Zhao Z., Chen Y., Cheng X., Huang L., Wen H., Xu Q., Zhou X., Zhang X., Chen J, & Ni T. (2023). The landscape of cryptic antisense transcription in human cancers reveals an oncogenic noncoding RNA in lung cancer. Science Advances, 9(14), eadf3264.

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Profiling Gene Expression in Lung Cell Lines

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The human normal lung epithelial cells named BEAS-2B was supplied by Beyotime Biotechnology (Hangzhou, China). The LUAD cell lines, including A-549 and NCI-H1299, were purchased from National Collection of Authenticated Cell Cultures (Shanghai, China). BEAS-2B and NCI-H1299 were cultured in 90% RPMI (Roswell Park Memorial Institute)-1640 with 10% FBS (fetal bovine serum). A-549 was cultured in 89% F-12K + 10% FBS + 1% Glutamax. We extracted the total RNA of the cell lines by RNAsimple Total RNA Kit (Tiangen, China). Whereafter, to acquire cDNA, we reverse transcribed the cell RNA that we have obtained applying PrimeScript RT reagent Kit (Takara, Otsu, Japan). Finally, based on the premixed system of 2 μL cDNA with SYBR Premix Ex Taq (Takara, Otsu, Japan) and primers, we detected the expression values of related genes in cell lines by Applied Biosystems StepOne Plus Real-Time PCR system (Life Technologies, Grand Island, NY, USA). The primers of the target gene were supplied by Sangon Biotech (Shanghai, China). The sequences of the primers used were listed in Table 1.

Peng H., Li X., Luan Y., Wang C, & Wang W. (2023). A novel prognostic model related to oxidative stress for treatment prediction in lung adenocarcinoma. Frontiers in Oncology, 13, 1078697.

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Cell Culture Conditions for NSCLC Lines

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The NSCLC cell lines NCI-H460 (H460) and NCI-H522 [H522; American Type Culture Collection (ATCC), Manassas, VA, USA] were grown in F-12K medium (Gibco; Thermo Fisher Scientific Inc., Waltham, MA, USA). The NSCLC cell lines A549 (ATCC) and NCI-H1299 (H1299; National Collection of Authenticated Cell cultures, Shanghai, China) were cultured in RPMI-1640 medium. Minimum essential medium was applied for culture of the NSCLC cell line SK-MES-1 (ATCC). BEAS-2B (ATCC), an immortalized human bronchial epithelial mesothelial cell line, was maintained in bronchial epithelial cell growth medium (Lonza, Walkersville, MD, USA). Meanwhile, 10% fetal bovine serum (FBS; Gibco) was used in the cultures of all of the above cell lines, which were cultivated at 37°C in a humidified atmosphere with 5% CO₂.

Chen G., Wang K., Li G., Wang L., Xiao Y, & Chen B. (2022). Long Noncoding RNA LAMTOR5-AS1 Interference Affects MicroRNA-506-3p/E2F6-Mediated Behavior of Non-Small Cell Lung Cancer Cells. Oncology Research, 28(9), 945-959.

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Culturing NSCLC and Normal Lung Cells

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The human NSCLC cell lines A549, NCI‐H1299, HCC827, NCI‐1650 and NCI‐H358 were procured from the National Collection of Authenticated Cell Cultures (Shanghai, China), and the human normal lung epithelial cell line DEAS‐2B was provided by Beyotime. A549 cells were cultured in DMEM/F12 medium (Procell) containing 10% fetal bovine serum (FBS, Gibco). BEAS‐2B, NCI‐H1299, HCC827, NCI‐1650 and NCI‐H358 cells were cultured in RPMI 1640 medium (Procell) containing 1% glutamax (Invitrogen), 1% (v/v) 100 mM sodium pyruvate solution (Invitrogen) and 10% FBS. All cells were maintained in a humidified chamber supplemented with 5% CO₂.

Zhu W., Shi L., Gong Y., Zhuo L., Wang S., Chen S., Zhang B, & Ke B. (2022). Upregulation of ADAMDEC1 correlates with tumor progression and predicts poor prognosis in non‐small cell lung cancer (NSCLC) via the PI3K/AKT pathway. Thoracic Cancer, 13(7), 1027-1039.

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Lung Cancer Cell Line Culture

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LUAD cell lines (NCI-H1299, NCI-H209, A549, and NCI-H524), combined with BEAS-2B, which is a human normal lung epithelial cell line, were purchased from the Cell Bank of the Chinese Academy of Sciences (Beijing, China). The Dulbecco’s Modified Eagle Medium containing 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA) was used for cell culture at 37°C and 5% CO₂.

Liu J., Zhang Y., Tao J., Yu T, & Zhang T. (2022). Heat shock factor 2-binding protein promotes tumor progression via activation of MAPK signaling pathway in lung adenocarcinoma. Bioengineered, 13(4), 10324-10334.

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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.

Pei X., Chen S.W., Long X., Zhu S.Q., Qiu B.Q., Lin K., Lu F., Xu J.J., Zhang P.F, & Wu Y.B. (2020). circMET promotes NSCLC cell proliferation, metastasis, and immune evasion by regulating the miR-145-5p/CXCL3 axis. Aging (Albany NY), 12(13), 13038-13058.

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NSCLC Tissue and Cell Line Collection

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A total of 90 pairs of NSCLC tissues and paired adjacent normal tissues were collected from Peking University People's Hospital (Beijing, China). All patients received lobectomy or wedge resection without any preoperative radiation or chemotherapy. The clinical data and follow‐up information were also collected. This study was approved by the Ethics Committee of Peking University People's Hospital. Each patient included in this study provided written informed consent.
NSCLC cell lines (A549, SPC‐A1, NCI‐H1703, NCI‐H1299, and H1975) and 16‐HBE cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). SPC‐A1 and 16‐HBE cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco) and others were cultured in RPMI 1640 medium (Gibco). The cell lines were identified by short tandem repeat (STR) and tested for mycoplasma contamination. All media were supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin. All cell lines were cultured at 37°C with 5% CO₂ in a humidified incubator.

Li H., Huang Q., Guo H., Chen X., Li X, & Qiu M. (2023). Circular RNA, circular RARS, promotes aerobic glycolysis of non‐small‐cell lung cancer by binding with LDHA. Thoracic Cancer, 14(4), 389-398.

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