C666-1 cells were purchased from Kalang Biomart, SUNE1 cells and human immortalized nasopharyngeal epithelial cells NP69 were purchased from Sbj-Bio Life Science. SUNE1, C666-1 and NP69 cells were maintained in RPMI-1640 medium (Gibco) supplemented with 10% FBS and 0.05% penicillin/streptomycin. Small interfering RNA (siRNA) targeting UCA1 (si-UCA1, including si-UCA1#1, si-UCA1#2 and si-UCA1#3), siRNA targeting ITGB1 (si-ITGB1), siRNA negative control (si-NC), pcDNA and pcDNA-ITGB1 overexpression vector (ITGB1) were synthesized by Genepharma. The miRNA mimic (miR-124-3p), miR-124-3p inhibitor (in-miR-124-3p), miRNA negative control (miR-NC) and miRNA negative control inhibitor (in-miR-NC) were purchased from RIBOBIO. SUNE1 and C666-1 cells were transfected with those plasmids using Lipofectamine 2000 (Invitrogen).
Sune1
Manufactured by Thermo Fisher Scientific
Sourced in United States
The SUNE1 is a laboratory equipment designed for scientific applications. It serves as a core component in various research and analytical processes. The device's primary function is to provide a controlled environment for specific tasks, though the exact intended use may vary depending on the application.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions)
C666 1, by Thermo Fisher Scientific (6 mentions)
Rpmi 1640, by Thermo Fisher Scientific (5 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Dmem medium, by Thermo Fisher Scientific (3 mentions)
Sune1, by American Type Culture Collection (2 mentions)
C666 1, by National Collection of Authenticated Cell Cultures (1 mentions)
Si nc, by Thermo Fisher Scientific (1 mentions)
Si snhg16, by GenePharma (1 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
Si nc, by GenePharma (1 mentions)
C666 1, by BNCC (1 mentions)
Keratinocyte serum free medium, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Hone1, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Cne 1, by Shanghai Institute of Biochemistry and Cell Biology (1 mentions)
Cne 2, by Shanghai Institute of Biochemistry and Cell Biology (1 mentions)
Inverted phase contrast microscope, by Olympus (1 mentions)
12 protocols using sune1
1
Exploring SUNE1, C666-1, and NP69 Cells
2
Nasopharyngeal Cell Line Transfection
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Two NPC cell lines (C666-1 and SUNE-1) and the normal nasopharyngeal cell line NP69 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in DMEM medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% FBS (Gibco).
siRNA-NC (5′-AAUUCUCCGAACGUGUCACGU-3′), siRNA-HOTTIP (5′-GCGUCUACAUUAACAAAGAUU-3′), miR-4301 mimics (5′-UCCCACUACUUCACUUGUGA-3′) and corresponding negative control (5′-UCACAACCUCCUAGAAAGAGUAGA-3′) were synthesized by RiboBio (Guangzhou, China). SUNE-1 cells were cultured in 6-well plates for 24 h and transfected with siRNA-HOTTIP, miR-4301 mimics, or their respective negative controls using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. To determine the efficiency of siRNA-HOTTIP, the expression of HOTTIP was assessed with real-time PCR.
siRNA-NC (5′-AAUUCUCCGAACGUGUCACGU-3′), siRNA-HOTTIP (5′-GCGUCUACAUUAACAAAGAUU-3′), miR-4301 mimics (5′-UCCCACUACUUCACUUGUGA-3′) and corresponding negative control (5′-UCACAACCUCCUAGAAAGAGUAGA-3′) were synthesized by RiboBio (Guangzhou, China). SUNE-1 cells were cultured in 6-well plates for 24 h and transfected with siRNA-HOTTIP, miR-4301 mimics, or their respective negative controls using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. To determine the efficiency of siRNA-HOTTIP, the expression of HOTTIP was assessed with real-time PCR.
3
Regulation of NPC Cells by SNHG16
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The NPC cell lines (SUNE1, 5–8F, and C666-1) and normal nasopharyngeal epithelial cell line (NP69) were purchased from BeNa (Beijing, China). The cells were incubated in DMEM with 10% FBS and kept in an incubator with 5% CO2 at 37°C.
The siRNA targeting SNHG16 (si-SNHG16: 5ʹ-ACAAAGUAUGACAGUUCGGCC-3ʹ), siRNA negative control (si-NC: 5ʹ-CUCUGCGUCAUCUAGAUGUGA-3ʹ), miR-520a-3p mimics (5ʹ-AAAGUGCUUCCCUUUGGACUG-3ʹ), miR-520a-3p inhibitor (5ʹ-UUUCACGAAGGGAAACCUGAC-3ʹ), and their corresponding negative controls (NC mimics: 5ʹ-ACGUAGUGAUCAGACCUGAAC-3ʹ and NC inhibitor: 5ʹ-UUCACUUGGCCAUAUUAGGGC-3ʹ) were acquired from GenePharma (Shanghai, China). The full length of MAPK1 was inserted into pcDNA3.1 to establish MAPK1 overexpression plasmid (MAPK1). With Lipofectamine 2000 (Invitrogen), si-NC, si-SNHG16, MAPK1, pcDNA3.1, miR-520a-3p, miR-520a-3p inhibitor, NC mimics, and NC inhibitor were transfected into SUNE1 and 5–8F cells.
The siRNA targeting SNHG16 (si-SNHG16: 5ʹ-ACAAAGUAUGACAGUUCGGCC-3ʹ), siRNA negative control (si-NC: 5ʹ-CUCUGCGUCAUCUAGAUGUGA-3ʹ), miR-520a-3p mimics (5ʹ-AAAGUGCUUCCCUUUGGACUG-3ʹ), miR-520a-3p inhibitor (5ʹ-UUUCACGAAGGGAAACCUGAC-3ʹ), and their corresponding negative controls (NC mimics: 5ʹ-ACGUAGUGAUCAGACCUGAAC-3ʹ and NC inhibitor: 5ʹ-UUCACUUGGCCAUAUUAGGGC-3ʹ) were acquired from GenePharma (Shanghai, China). The full length of MAPK1 was inserted into pcDNA3.1 to establish MAPK1 overexpression plasmid (MAPK1). With Lipofectamine 2000 (Invitrogen), si-NC, si-SNHG16, MAPK1, pcDNA3.1, miR-520a-3p, miR-520a-3p inhibitor, NC mimics, and NC inhibitor were transfected into SUNE1 and 5–8F cells.
4
Culturing Human Nasopharyngeal Cell Lines
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The human NPC cell lines 6–10B and SUNE1 were obtained from Shanghai Biotech Bingsui Co. Ltd. (Shanghai, China). The human NPC cell line C666-1 and the normal nasopharyngeal cell line NP69 were purchased from BeNa Culture Collection (BNCC, China). The cell lines 6–10B, SUNE1 and C666-1cells were maintained at 37 °C in RPMI-1640 medium, 5–8F cells in DMEM medium, and NP69 in keratinocyte serum-free medium (Invitrogen, Carlsbad CA USA) in a 5% CO2 incubator.
5
Nasopharyngeal Carcinoma Cell Line Knockdown
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The NPC cell lines (CNE2, SUNE1 and HONE1) were purchased from American Type Culture Collection (ATCC) (ATCC, Manassas, USA) and used within two months after resuscitation of frozen aliquots. The immortalized nasopharyngeal epithelial cell (NPEC2 Bmi-1) [42] (link) was kindly provided by Prof. Mu-Sheng Zeng. The NPC cell lines (CNE2, SUNE1 and HONE1) were cultured in RPMI 1640 with 10% fetal bovine serum (Invitrogen, CA, USA). The immortalized nasopharyngeal epithelial cell, NPEC2 Bmi-1 was cultured in KSF (Invitrogen, CA, USA). All the cell lines were grown in a humidified incubator at 37°C with 5% CO2.
CNE2 and SUNE1 cells were plated in 6-well dishes at 2×105 cells/well. Knockdown experiments were done after 24 hours seeding. LNA-control or LNA-antimiR-214 (Exiqon A/S, Vedbaek, Denmark) 50 nmol were transfected into cells using reagent of Lipofectamine RNAiMAX (Invitrogen, CA, USA) according to the manufacturer's instruction.
LNA-antimiR-214 sequence was5′-TGCCTGTCTGTGCCTGCTG-3′ .
CNE2 and SUNE1 cells were plated in 6-well dishes at 2×105 cells/well. Knockdown experiments were done after 24 hours seeding. LNA-control or LNA-antimiR-214 (Exiqon A/S, Vedbaek, Denmark) 50 nmol were transfected into cells using reagent of Lipofectamine RNAiMAX (Invitrogen, CA, USA) according to the manufacturer's instruction.
LNA-antimiR-214 sequence was
6
Establishing Cisplatin-Resistant NPC Cell Lines
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The NP-69 human normal nasopharyngeal epithelial cell line (cat. no. BNCC338439), and the 5–8F (cat. no. BNCC353307) and SUNE1 (cat. no. BNCC342441) human NPC cell lines were purchased from BeNa Culture Collection (Beijing Beina Chunglian Biotechnology Research Institute). 5–8F and SUNE1 cells were cultured in RPMI-1640 medium (Invitrogen; Thermo Fisher Scientific, Inc.) containing streptomycin (100 U/ml) and 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C in a humidified 5% CO2 environment. DDP-resistant NPC (5-8F/DDP and SUNE1/DDP) cell lines were established by continuous exposure to progressively increasing DDP (Sigma Aldrich; Merck KGaA) concentrations (0.5, 2, 4, 6 and 8 µg/ml) every 4 weeks. DDP-resistant NPC cells were cultured in RPMI-1640 medium supplemented with 2 µg/ml DDP and 10% FBS at 37°C in a humidified incubator with 5% CO2.
7
Cultivation of Human NPC Cell Lines
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Human NPC cell lines (HK1, 5‐8F, C666, SUNE1, 6‐10B) and nasopharyngeal epithelial cell line (NP69) were purchased from Shanghai institute of biological sciences, Chinese academy of sciences (Shanghai, China), which were cultivated by RPMI‐1640 enriched with 10% fetal bovine serum (FBS, Gibco, USA) in a 5% CO2 atmosphere at 37°C.
8
Nasopharyngeal Carcinoma Cell Line Culture
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Three nasopharyngeal carcinoma cell lines (CNE-1, CNE-2, and SUNE-1) were obtained from the Shanghai Institute of Cell Biology (Shanghai, China). CNE-1, CNE-2, and SUNE-1 cell lines were cultured in RPMI 1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (GE Healthcare Life Sciences, Logan, UT, USA) at 37°C with humidified 5% CO2.
9
Evaluating NVP-BEZ235 and Rapamycin Effects on SUNE1 Cells
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The NPC cell line SUNE1 was obtained from Wuxi Innovate Biomedical Technology Co., Ltd. (Wuxi, China). SUNE1 cells were incubated with Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) at 37°C. SUNE1 cells were seeded into a 24-well plate at the density of 80%, and then were incubated with 100 nM NVP-BEZ235 (cat. no. A506167) (14 (link)), 100 nM Rapamycin (cat. no. A606203; both Sangon Biotech Co., Ltd., Shanghai, China) (15 (link)), or 100 nM NVP-BEZ235 and 100 nM Rapamycin for 48 h at 37°C. The control group was administered the same amount of dimethyl sulphoxide (DMSO; Sangon Biotech Co., Ltd.). Cell morphology following treatment was observed with an inverted phase contrast microscope (Olympus Corporation, Tokyo, Japan; magnification, ×100).
10
Nasopharyngeal Cell Culture Protocols
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The nasopharyngeal epithelial cell line, NP-69 and NPC cells (SUNE-1, CNE-2, CNE-1 and C666-1) were cultured with RPMI-1640 supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA). C666-1 and SUNE-1 cells are not misidentification and contamination of human cell lines (ExPASy, https://www.expasy.org/ ). All cell lines were obtained from KeyGen Biotech Co., Ltd (Nanjing, Jiangsu, China) and maintained at 37°C in a humid chamber with 5% CO2.
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