Nci h1299

Manufactured by Thermo Fisher Scientific

Sourced in United States, China

The NCI-H1299 is a laboratory cell line derived from a human non-small cell lung carcinoma. It is commonly used in cancer research and drug development studies.

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57 protocols using nci h1299

Comprehensive Cell Line Validation and Inhibitor Characterization

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All cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), independently validated by short tandem repeat (STR) DNA fingerprinting, and tested negative for mycoplasma infection. NCI-H157, A549, NCI-H1299, HeLa, BT549, MDA-MB-231, CT26, B16-F10, MC-38, LLC and 293T cells were cultured in RPMI 1640 or DMEM media, supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C in a humidified atmosphere with 5% CO₂. Inhibitors for ATM (KU-60019, AZD1390, AZD0156), STING (H151), p-TBK1 (GSK8612), p-STAT1 (fludarabine) were purchased from Selleck (Houston, TX, USA). Human IFNβ and Galectin-9 ELISA assay Kits were purchased from R&D SYSTEMS (Minneapolis, MN, USA). Anti-human Gal-9 antibody was purchased from BioRad (Hercules, CA, USA). Anti-mouse Gal-9 antibody and IgG isotype control were purchased from BioXCell (Lebanon, NH, USA). Antibodies against cGAS, STING, p-STING(Ser366), p-TKB1(Ser172), TKB1, p-STAT1(Tyr701), STAT1 and Actin were purchased from Cell Signaling Technology (Cambridge, MA, USA). Anti-mouse fluorochrome-conjugated antibodies including FITC-CD4, Percp/Cy5.5-CD8, AF700-CD3, APC/Cy7-CD45, PE/Cy7-IFNγ antibodies were purchased from BioLegend (San Diego, CA, USA).

Zheng S., Song J., Linghu D., Yang R., Liu B., Xue Z., Chen Q., Liu C., Zhong D., Hung M.C, & Sun L. (2023). Galectin-9 blockade synergizes with ATM inhibition to induce potent anti-tumor immunity. International Journal of Biological Sciences, 19(3), 981-993.

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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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Cell Culture Conditions for Diverse Cell Lines

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A549, SK-MES-1, NCI-H1299, and 293T cells were acquired from Cobioer (Jiangsu, People's Republic of China). SK-MES-1 cells were cultured in MEM medium supplemented with 10% (v/v) heat-inactivated newborn calf serum (Thermo Fisher Scientific) with 1 ×10⁵ U/L penicillin G and 1 ×10⁵ U/L streptomycin in a humidified incubator at 37°C and 5% CO₂. NCI-H1299, A549, and A549/control-shRNA cells were cultured in RPMI l640 medium supplemented with 10% (v/v) heat-inactivated newborn calf serum (Thermo Fisher Scientific) with 1 ×10⁵ U/L penicillin G and 1 ×10⁵ U/L streptomycin in a humidified incubator at 37°C and 5% CO₂. 293T cells were cultured in DMEM medium supplemented with 10% (v/v) heat-inactivated newborn calf serum (Thermo Fisher Scientific) with 1 ×10⁵ U/L penicillin G and 1 ×10⁵ U/L streptomycin in a humidified incubator at 37°C and 5% CO₂.

Zhang H., Chen R., Wang X., Zhang H., Zhu X, & Chen J. (2019). Lobaplatin-Induced Apoptosis Requires p53-Mediated p38MAPK Activation Through ROS Generation in Non-Small-Cell Lung Cancer. Frontiers in Oncology, 9, 538.

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Lung Cell Lines for Cancer Research

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A549 (EGFR-WT, KRAS mutation), NCI-H1299 (EGFR-WT, KRAS-WT), and NCI-H226 (EGFR-WT, KRAS-WT) cells were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA). NCI-H1437 (EGFR-WT, KRAS-WT) and BEAS-2B (normal human bronchial epithelial cell line) cells are kind gifts from Professor Ming-Jen Hsu at the Department of Pharmacology at Taipei Medical University. A549, NCI-H1299, NCI-226, and NCI-H1437 cells were maintained in 10% fetal bovine serum (FBS)-supplemented RPMI 1640 medium and 1 × antibiotic–antimycotic (Thermo Fisher Scientific, Waltham, MA, USA), and BEAS-2B cells were maintained in bronchial epithelial growth medium (Lonza, Walkersville, MD, USA) at 37 °C in a humidified incubator containing 5% CO₂. Cucurbitacin E was purchased from MedChem Express (Monmouth Junction, NJ, USA).

Hsu H.L., Lin B.J., Lin Y.C., Tu C.C., Nguyen N.L., Wang C.C., Chen M.C, & Chen C.H. (2023). Cucurbitacin E Exerts Anti-Proliferative Activity via Promoting p62-Dependent Apoptosis in Human Non-Small-Cell Lung Cancer A549 Cells. Current Issues in Molecular Biology, 45(10), 8138-8151.

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Culturing Human Cell Lines for Research

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Human bronchial epithelial cells (HBEpiC), human lung cancer cell lines (A549 cells and NCI‐H1299 cells) and human myeloid leukaemia mononuclear cells (THP‐1) were obtained from American Type Culture Collection (ATCC). NCI‐H1299 and THP‐1 were cultured in RPMI 1640 supplemented with 10% FBS (Thermo Scientific, Waltham, Massachusetts, USA). HBEpiC and A549 were cultured in HycloneDME‐F12 supplemented with 10% FBS. All cells were cultured in 5% CO2 at 37°C. LPS and recombinant protein IL‐4 were from Beyotime Biotechnology (Nanjing, China).

Wang Y., Miao Z., Qin X., Li B, & Han Y. (2021). NOD2 deficiency confers a pro‐tumorigenic macrophage phenotype to promote lung adenocarcinoma progression. Journal of Cellular and Molecular Medicine, 25(15), 7545-7558.

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Lentiviral Transduction of Lung Cancer Cells

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The lentiviral vector with NFIX plasmids (shRNA) was purchased from Thermo Scientific Open Biosystems (BD Biosciences, United States). Plasmids were cultured in Luria broth with ampicillin (AMRESCO, United States) and DNA was extracted using Qiagen purification kit (Germany) for transient transfection. Two human lung cancer cell lines, A549 and NCI-H1299 were purchased from American Type Culture Collection (United States). A549 cells, from a human lung carcinoma cell, maintained in Kaighn’s modification of Ham’s F-12 (F-12K) medium (Thermo Fisher Scientific, United States). NCI-H1299 cells, also derived from a human lung carcinoma and metastatic site lymph nodes, maintained in Roswell Park Memorial Institute1640 (RPMI1640) medium (Thermo Fisher Scientific, United States). Both medium were supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, United States).

Rahman N.I., Abdul Murad N.A., Mollah M.M., Jamal R, & Harun R. (2017). NFIX as a Master Regulator for Lung Cancer Progression. Frontiers in Pharmacology, 8, 540.

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Cultivation of NSCLC Cell Lines

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NSCLC cell lines, including the NCI-H1299 (adenocarcinoma) and SK-MES-1 (squamous carcinoma) cell lines were purchased from the Cell Resource Center, Shanghai Institute of Biotechnology, Chinese Academy of Sciences. NCI-H1299 and SK-MES-1 cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Inc.) with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc.) and minimum essential medium (Thermo Fisher Scientific, Inc.) with 10% FBS, respectively. Both cell lines were incubated in an incubator with 5% CO₂ at 37°C.

Xie K., Ni X., Lv S., Zhou G, & He H. (2021). Synergistic effects of olaparib combined with ERCC1 on the sensitivity of cisplatin in non-small cell lung cancer. Oncology Letters, 21(5), 365.

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Cell Culture Protocol for NSCLC Cell Lines

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Normal lung epithelial cells (BEAS2B) and NSCLC cell lines A549 and NCI-H1299 were obtained from Procell Life Science & Technology Co., Ltd. A549 and NCI-H1299 cells were maintained in complete medium consisting of DMEM (Thermo Fisher Scientific), 0.1% penicillin‒streptomycin, and 10% fetal bovine serum (FBS, Thermo Fisher Scientific, category number: 26010-074) at 37°C in a humidified atmosphere of 5% CO₂. Cells were maintained in a 75 cm² culture flask for culture. Cells were subcultured every 3 days with a maximum of passages not exceeding 25. When the cells grew to 70% confluence, they were collected and used for subsequent experiments.

Liu J., Chen L., Zhang J., Luo X., Tan Y, & Qian S. (2023). AS-IV enhances the antitumor effects of propofol in NSCLC cells by inhibiting autophagy. Open Medicine, 18(1), 20230799.

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Culturing Lung Cell Lines

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Normal human bronchial epithelial cells (HBEs) and human non-small cell lung cancer (NSCLC) cell lines, A549, NCI-H1299, NCI-H1650, and NCI-H460, were obtained from Invitrogen (San Mateo, CA, USA). Cells were cultured in RPMI 1640 medium containing 20% fetal bovine serum (FBS) and incubated at 37°C and 5% CO₂. When cell confluence reached 80%, cells were digested with 0.25% trypsin and sub-cultured.

Xu X., Jiang J., Yao L, & Ji B. (2018). Silencing the FOLR2 Gene Inhibits Cell Proliferation and Increases Apoptosis in the NCI-H1650 Non-Small Cell Lung Cancer Cell Line via Inhibition of AKT/Mammalian Target of Rapamycin (mTOR)/Ribosomal Protein S6 Kinase 1 (S6K1) Signaling. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 24, 8064-8073.

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Establishing NSCLC Cell Lines for Research

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Human NSCLC cell lines [human bronchial epithelial (HBE), National Cancer Institute (NCI)-H1299, A549, NCI-H1650, NCI-H2228 and NCI-H292] were purchased from American Type Culture Collection (MA, USA). The cell lines were frozen and stored in our laboratory. The cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco Company, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS, HyClone Company, Logan, UT, USA) and 1% Penicillin-Streptomycin (Life Technologies, Paisley, UK) in an incubator with 5% CO₂ at 37 °C.
Short hairpin RNAs (shRNAs, sh-Scrambled, and 3 shRNAs for TRPM2-AS), miRNA mimics, and the corresponding controls (listed in Table 1, GenePharma, Shanghai, China) were transfected into NCI-H1299 and A549 cells with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) in line with the manufacturer’s protocol. Stable cell lines were obtained by using 2 µg/mL puromycin (Sigma-Aldrich, St-Louis, Missouri, USA) for 3 weeks to eliminate untransfected cells. TRPM2-AS 875 bp cDNA sequences were subcloned and amplified into the lentiviral expression vector (GenePharma).

Cui D., Feng Y., Shi K., Zhang H, & Qian R. (2020). Long non-coding RNA TRPM2-AS sponges microRNA-138-5p to activate epidermal growth factor receptor and PI3K/AKT signaling in non-small cell lung cancer. Annals of Translational Medicine, 8(20), 1313.

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