Human colon epithelial cells NCM460 and CRC cell lines (including HCT116, HT29, and SW620) were purchased from Procell Life Science & Technology Co., Ltd. (Procell, Wuhan, China). NCM460 and SW620 cells were cultured in DMEM medium (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA) and 1% penicillin–streptomycin (100 U/mL penicillin and 100 μg/mL streptomycin). HCT116 and HT29 cells were cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA) and 1% penicillin/streptomycin (Gibco). SLC35A3 overexpression models were established in HCT116 and SW620 cells using the pcDNA3.1( +)-SLC35A3 plasmid with the assistance of Lipofectamine 8000 transfection reagent (Beyotime). Empty vector was used as a negative control. After 48 h of cell culture, the corresponding phenotypic experiments were performed. The pcDNA3.1( +) plasmid containing SLC35A3 coding sequence was constructed by GenePharma (Shanghai, China). Primers for SLC35A3 (forward, 5′-GCTTGGTACCGAGCTCGGATCCG-3′, reverse, 5′-TGCTGGATATCT GCAGAATTCCTATGCTTTAGTGGGATTTCCTGCAGG-3′) were provided by GenePharma.
Hct116
Manufactured by Beyotime
Sourced in China
The HCT116 is a cell line derived from human colorectal carcinoma. It is a commonly used model for cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Hct116, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Sw620, by Thermo Fisher Scientific (2 mentions)
Hct116, by American Type Culture Collection (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Pcdna3.1 plasmid, by GenePharma (1 mentions)
Ncm460, by Thermo Fisher Scientific (1 mentions)
Ht 29, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 8000 transfection reagent, by Beyotime (1 mentions)
Ncm460, by Procell (1 mentions)
Sw620, by Procell (1 mentions)
Hct116, by Procell (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Sgc 7901, by Beyotime (1 mentions)
Rpmi 1640, by Beyotime (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dld 1, by American Type Culture Collection (1 mentions)
Penicillin g, by Beyotime (1 mentions)
5 protocols using hct116
1
Colon Cancer Cell Lines: SLC35A3 Overexpression
2
Cancer Cell Culture Protocols
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Human cancer cell lines HepG2, SGC-7901, HCT-116, A549, NCI-H1299 and MCF-7 as well as normal liver cell line L02 were purchased from the Cell Bank of Shanghai Institute of Cell Biology. Among them, MCF-7, HepG2 cells were cultured at 37 °C in 5% CO2 with DMEM supplemented with 10% fetal bovine serum (FBS) (Hyclone, Lifescience, USA) and 1% penicillin/streptomycin (Beyotime, Nantong, China), while HCT-116, SGC-7901, A549, NCI-H1299 and L02 cells were cultured at 37 °C in 5% CO2 with RPMI-1640 supplemented with 10% FBS and 1% penicillin/streptomycin. Cells were passed every 2 days and restarted from frozen stocks upon reaching pass number 20.
3
Culturing of Colorectal Cell Lines
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HCT116 and DLD-1 cell lines were purchased from American Type Culture Collection [(ATCC) Manassas, VA, United States]. The HCT116 and DLD-1 cells were cultured in Dulbecco’s modified Eagle medium in an incubator with 5% CO2 at 37° C. Penicillin G (100 U/mL), streptomycin (100 mg/mL) (Beyotime, Nanjing, China), and 10% foetal bovine serum (Invitrogen, Waltham, MA, United States) were added to the culture medium. Two colonic cell lines were used to demonstrate reproducibility.
4
Evaluating RPMTG's Effects on CRC Cell Lines
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The human CRC cell lines HCT116 and SW620 were purchased from The Cell Bank of Type Culture Collection of The Chinese Academy of Sciences. HCT116 and SW620 cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc.), supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin. The HCT116 (1×104 cells/well) and SW620 (2×104 cells/well) cells were pretreated with JNK inhibitor (SP600125, 10 µM; Beyotime Institute of Biotechnology) and p38 inhibitor (SB203580, 10 µM; Beyotime Institute of Biotechnology) for 2 h, and then treated with 250 µg/ml RPMTG for 24 h in a 37°C incubator containing 5% CO2. In subsequent experiments, the effects of RPMTG combined with JNK inhibitors or p38 inhibitors on proliferation and apoptosis were evaluated.
5
Colorectal Cancer Cell Lines Cultured
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CRC lines SW480 and HCT116 purchased from American type culture collection (ATCC; Manassas, VA, United States) were cultured in Dulbecco’s Modified Eagle Medium (HyClone, Logan, UT, United States) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin antibiotics at 37°C with 5% CO2. CRC line HT29 was from ATCC and cultured in McCoy’s 5A Medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. The control colorectal cell line NCM460 was purchased from BeNa Culture Collection (BNCC, Beijing, China) and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. When the cells reached a density of 45–50%, EA (MedChemExpress, Monmouth Junction, NJ, United States) was added to the medium at different concentrations. The EA was solved into DMSO (Solarbio life science, Beijing, China). The Cell Counting Kit-8 (CCK8; Beyotime, Shanghai, China) was used to detect the inhibitory effect of EA on the activity of SW480 and HCT116, according to the manufacturer’s instructions.
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