Six human colorectal cancer cell lines HCT116, SW480, HT29, SW620, Caco2 and LOVO were obtained from the Shanghai Institute of Cell Biology (Shanghai, China).The related oxaliplatin-resistant cell lines SW480-OxR and HCT116-OxR were generated by continuous exposure to increasing concentrations of oxaliplatin for a 10-month period as described previously 23 (link). We performed cytotoxicity testing to confirm that chemoresistance could be stable for about 4 weeks without oxaliplatin exposure. The oxaliplatin-resistant cell lines were used at no higher than 15 passages from creation. All cells were cultured in the appropriate medium (RPMI-1640 for HT29, SW620, SW480 and SW480-OxR cells; DMEM for Caco2, LOVO, HCT116 and HCT116-OxR cells) supplemented with 10% FBS (Gibco, Carlsbad, CA, USA) in a humidified atmosphere with 5% CO2 at 37 °C.
Sw480
Manufactured by Shanghai Institute of Biochemistry and Cell Biology
Sourced in China
The SW480 is a cell line derived from a human colorectal adenocarcinoma. It is commonly used in research to study the biology and pathogenesis of colorectal cancer.
Automatically generated - may contain errors
Lab products found in correlation
Hct116, by Shanghai Institute of Biochemistry and Cell Biology (12 mentions)
Sw620, by Shanghai Institute of Biochemistry and Cell Biology (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Sw480, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Thp 1, by Beyotime (3 mentions)
Phorbol 12 myristate 13 acetate pam, by Beyotime (3 mentions)
Thp 1, by Shanghai Institute of Biochemistry and Cell Biology (3 mentions)
Hct116, by Thermo Fisher Scientific (2 mentions)
Ht 29, by Shanghai Institute of Biochemistry and Cell Biology (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Ccl244, by Shanghai Institute of Biochemistry and Cell Biology (2 mentions)
Dld 1, by Shanghai Institute of Biochemistry and Cell Biology (2 mentions)
Caco 2, by Thermo Fisher Scientific (1 mentions)
Sw620, by Thermo Fisher Scientific (1 mentions)
Ht 29, by Thermo Fisher Scientific (1 mentions)
Caco 2, by Shanghai Institute of Biochemistry and Cell Biology (1 mentions)
18 protocols using sw480
1
Establishing Oxaliplatin-Resistant Colorectal Cancer Cell Lines
2
Colorectal Cancer Tissue Profiling
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Clinical specimens. fifty-three cases of fresh colorectal cancer (CRC) tissues and matched adjacent normal colorectal tissues were collected immediately after surgical resection and stored at -80˚C for further analysis. This study was conducted in accordance with the Declaration of Helsinki and with approval from the Ethics Committee of Soochow University. Table I shows the clinical characteristics of these patients.
Cell cultures and irradiation. All the cell lines (fHC, CCL244, HCT116, SW480 and LOvO) were purchased from Shanghai Institute of Cell Biology (Shanghai, China) and maintained in DMEM supplemented with 10% fBS and antibiotics (100 U/ml penicillin G, 100 U/ml streptomycin sulfates; Gibco, Grand Island, NY, USA) at 37˚C in a humidified atmosphere containing 5% CO 2 . Cells were exposed to a single dose of X-ray irradiation from the linear accelerator (RadSource, Suwanee, GA, USA) at a dose rate of 1.15 Gy/min.
SiRNA construction and transfection. The siRNA sequences targeting human HOTAIR (si-HOTAIR) or negative control (NC) sequence were designed and synthesized by Genepharma (Shanghai, China) as shown in Table II. Cells were transfected with siRNA by Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).
Cell cultures and irradiation. All the cell lines (fHC, CCL244, HCT116, SW480 and LOvO) were purchased from Shanghai Institute of Cell Biology (Shanghai, China) and maintained in DMEM supplemented with 10% fBS and antibiotics (100 U/ml penicillin G, 100 U/ml streptomycin sulfates; Gibco, Grand Island, NY, USA) at 37˚C in a humidified atmosphere containing 5% CO 2 . Cells were exposed to a single dose of X-ray irradiation from the linear accelerator (RadSource, Suwanee, GA, USA) at a dose rate of 1.15 Gy/min.
SiRNA construction and transfection. The siRNA sequences targeting human HOTAIR (si-HOTAIR) or negative control (NC) sequence were designed and synthesized by Genepharma (Shanghai, China) as shown in Table II. Cells were transfected with siRNA by Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).
3
Colon Cancer Cell Line Transfection Protocol
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The human colon cancer cell lines SW480, LoVo, DLD-1 and HCT-116 were purchased from the Shanghai Institute of Cell Biology, China. The LS174T cell line was purchased from the ATCC. These cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA) and antibiotics at 37 °C with 5% CO2. Colon cancer cells were transfected with either siZNF460 or empty vector siRNA (Ribo, China) using a riboFETCTM CP transfection reagent (Ribo, China) according to the manufacturer's instructions. The siRNA-transfected colon cancer cells were cultivated at 37 °C for 48 hours. The transfected cells were divided into three experimental groups: Control, NC-siRNA, ZNF460siRNA. After 48 hours, the transfected cells were collected for protein expression analysis. The siRNA sequence is listed as follows: ZNF460siRNA#1: 5′-GCACAGATCTCATTCAAC-3′; ZNF460siRNA#2: 5′-GTCCCAAGATACTCCTATT-3′; ZNF460siRNA#3: 5′- GCCTTCAATTGCCGCTCAT-3′.
4
Culturing and Authenticating Human CRC Cell Lines
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The human CRC cell lines SW480, SW620, LOVO, DLD-1 and HCT-116 were purchased from the Shanghai Institute of Cell Biology. The cells were cultured in RPMI 1640 medium (Gibco, Grand Island, NY, USA) supplemented with fetal bovine serum (HyClone, Logan, UT, USA) to a final concentration of 10% and antibiotics at 37°C with 5% CO2. Plasmids and reagents are described in the Supplementary Materials and Methods . All these cells were authenticated using short tandem repeat profiling by the Cell Bank.
5
Colon Cancer Tissue Collection and Cell Culture
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We collected tumor samples and distant normal mucous tissues (>5 cm from the margin of the tumor) from 21 patients with colon cancer who received surgical resection at our hospital from January 2015 to December 2016. No patient had received neoadjuvant chemotherapy or chemotherapy prior to surgery, and all tissue samples were pathologically confirmed. Tumor-node-metastasis staging of the patients was according to the 2015 National Comprehensive Cancer Network clinical practice guidelines. Informed consent was obtained from each patient from the archives of the Central Hospital of Panyu District Hospital and with approval from the institutional ethics committee (Guangzhou, China).
Cell culture FIHC, HCT-116, HT-29, and SW480 cell lines were purchased from the Shanghai Institute of Cell Biology (Shanghai, China). The cell lines were cultured in Dulbecco's Modified Eagle's Medium (DMEM)/Nutrient Mixture F-12 Ham containing 10% fetal bovine serum, and maintained in a humidified incubator at 37°C with 5% CO 2 . The cells in logarithmic growth with 95% viability were subjected to further experiment.
Cell culture FIHC, HCT-116, HT-29, and SW480 cell lines were purchased from the Shanghai Institute of Cell Biology (Shanghai, China). The cell lines were cultured in Dulbecco's Modified Eagle's Medium (DMEM)/Nutrient Mixture F-12 Ham containing 10% fetal bovine serum, and maintained in a humidified incubator at 37°C with 5% CO 2 . The cells in logarithmic growth with 95% viability were subjected to further experiment.
6
Culturing Colon Cancer Cell Lines
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The SW480 and HCT116 colon cancer cell lines were purchased from the Shanghai Institute of Cell Biology (Shanghai, China). SW480 cells were cultured in Leibovitz's L-15 Medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37˚C in a humidified atmosphere containing 5% CO2. HCT116 cells were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37˚C in a humidified atmosphere containing 5% CO2.
7
Culturing Human Colon Cancer Cell Lines
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Human colon cancer cell lines HCT116 and SW480 were obtained from Shanghai Institute of Cell Biology, Chinese Academy of Sciences (Shanghai, China) and maintained in RPMI 1640 medium (Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine (Gibco, Carlsbad, CA, USA), 100 units/mL penicillin and 100 mg/mL streptomycin (Gibco, Carlsbad, CA, USA) at 37 °C in incubator containing 5% CO2. SFN and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, MO, USA).
8
Comparative Analysis of Colon Cancer Cell Lines
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The human colon cancer cell line SW480 and human oxaliplatin-resistant colon cancer cell line SW480-OXA were purchased from the Shanghai Institute of Cell Biology of the Chinese Academy of Sciences (Shanghai, China), and the normal human colon epithelial cell line NCM460 was obtained from INCELL. The mouse colon cancer cell line CT26 was purchased from the Shanghai Institute of Cell Biology of the Chinese Academy of Sciences (Shanghai, China). CT26, SW480, and SW480-OXA cells were cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco) with routine addition of ampicillin and streptomycin. NCM460 cells were cultured in DMEM/high-glucose medium (HyClone, GE Healthcare Life Sciences) supplemented in the same manner as for RPMI 1640.
9
Overexpression of miR-1 in Colorectal Cancer
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A normal intestinal epithelial cell line (FHC) and established human colorectal carcinoma cell lines (HCT116, CCL244, SW480, HT29 and Lovo) were obtained from the Shanghai Institute of Cell Biology (Shanghai, China) and were cultured in DMEM (HyClone, Logan, UT, USA), supplemented with 10% fetal bovine serum (FBS; HyClone) and 1% Penicillin-Streptomycin solution (Beyotime, Shanghai, China). All cell lines were incubated at 37 °C in a humidified incubator with a 5% CO2 atmosphere.
The cells were seeded into 6-well plates and transiently transfected with Lipofectamine 2000 (Invitrogen, San Diego, CA, USA) and miR-1 mimics or the miRNA mimic Negative Control (GenePharma, Shanghai, China), according to the manufacturer's instructions. Real-time PCR was performed to confirm the efficiency of transfection.
The cells were seeded into 6-well plates and transiently transfected with Lipofectamine 2000 (Invitrogen, San Diego, CA, USA) and miR-1 mimics or the miRNA mimic Negative Control (GenePharma, Shanghai, China), according to the manufacturer's instructions. Real-time PCR was performed to confirm the efficiency of transfection.
10
Modulating CRC Progression via CPEB3
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The human monocyte cell line THP-1 and CRC cell lines (SW480, HCT116, LoVo, and RKO) were obtained from the Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). Lentiviruses carrying fulllength CPEB3 or short hairpin RNA (shRNA_CPEB3) sequences targeting against human CPEB3 mRNA and matched negative controls were constructed by the Shanghai Institute of Biochemistry and Cell Biology. SW480, HCT116, LoVo and RKO cells were transfected with the indicated lentivirus overnight, then 2 μg/mL puromycin was added after 72 h of transfection to obtain stably transfected CRC cells. For macrophage generation, THP-1 cells were treated with 100 ng/mL phorbol-12-myristate-13-acetate (PAM) (Beyotime, Shanghai, China) for 12 h to differentiate into adhered macrophages. To obtain TAM supernatants, CRC cells were seeded in 0.4-μm pore inserts, then transferred to a 6-well plate seeded with THP-1 macrophages in advance and co-cultured for another 24 h. For co-culture experiments, stably transfected CRC cells were co-cultured with THP-1 macrophages for another 24 h.
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