cell culture human colon cancer cell lines  (ATCC)

Journal: Biomolecules & biomedicine

Article Title: Jun-APOE-LRP1 axis promotes tumor metastasis in colorectal cancer.

doi: 10.17305/bb.2023.9248

Figure Lengend Snippet: Figure 2. APOE-overexpression promotes CRC cell migration and invasion in vitro. The efficiency of APOE-overexpression in HCT116 and HCT8 cells were validated by qPCR (A) and western blot (B); (C) The cell proliferation curves of HCT116 and HCT8 cells with or without APOE-overexpression evaluated byCCK-8assay;(D)Representativeimagesofcolonyformationassays;(E)Quantitativeresultsofcolonyformationassays.Thecolonynumberswerecounted using ImageJ; (F) Representative images of migration and invasion assays of HCT116 and HCT8 cell lines in the presence or absence of APOE-overexpression; (G) Quantitative results of migration and invasion assays. The migrated and invaded cells were counted using ImageJ. apoE: Apolipoprotein E; CCK-8: Cell counting Kit-8; CRC: Colorectal cancer; OD: Optical density; OE: Overexpression; qPCR: Quantitative polymerase chain reaction; VC: Vector control; ns: No significance.

Article Snippet: Construction of APOE-overexpressing stable cell lines and cell culture Human colon cancer cell lines HCT116 and HCT8 (American Type Culture Collection, USA) were maintained in RPMI 1640 (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Gibco).

Techniques: Over Expression, Migration, In Vitro, Western Blot, CCK-8 Assay, Cell Counting, Real-time Polymerase Chain Reaction, Plasmid Preparation, Control

Journal: Biomolecules & biomedicine

Article Title: Jun-APOE-LRP1 axis promotes tumor metastasis in colorectal cancer.

doi: 10.17305/bb.2023.9248

Figure Lengend Snippet: Figure 3. Jun activates APOE-expression in CRC cells. (A) Comparison of JUN mRNA levels between normal and tumor samples. Data were obtained from the UCSC Xena database; (B) Representative ATAC-seq and ChIP-seq signals in the APOE promoter region. The ATAC-seq data of CRC patients were downloaded from the TCGA data portal. ChIP-seq data for H3K4me3, H3K27ac, and JUN were downloaded from the Cistrome DB database; (C and D) qPCR and western blot were performed to determine the APOE mRNA and protein levels following knockdown of JUN in HCT116 and HCT8 cell lines; (E) HCT116 and HCT8 cells were cotransfected with pGL4.10-APOE, pcDNA3.1-JUN (or pcDNA3.1 vector), and pRL-TK, and the luciferase activity was determined 24 h post-transfection. Firefly luciferase/Renilla luciferase was used to indicate promoter activity. apoE: Apolipoprotein E; ATAC-seq: Assay for targeting accessible chromatin with high-throughput sequencing; ChIP-seq: Chromatin immunoprecipitation sequencing; COAD: Colon adenocarcinoma; CRC: Colorectal cancer; TCGA: The Cancer Genome Atlas; qPCR: Quantitative polymerase chain reaction.

Article Snippet: Construction of APOE-overexpressing stable cell lines and cell culture Human colon cancer cell lines HCT116 and HCT8 (American Type Culture Collection, USA) were maintained in RPMI 1640 (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Gibco).

Techniques: Expressing, Comparison, ChIP-sequencing, Western Blot, Knockdown, Plasmid Preparation, Luciferase, Activity Assay, Transfection, Next-Generation Sequencing, Real-time Polymerase Chain Reaction

Journal: Biomolecules & biomedicine

Article Title: Jun-APOE-LRP1 axis promotes tumor metastasis in colorectal cancer.

doi: 10.17305/bb.2023.9248

Figure Lengend Snippet: Figure 4. APOE-overexpression reverses the metastasis suppression of JUN knockdown. (A) Following knockdown of JUN, HCT116, and HCT8 cells were transfected with pLV-APOE (pLV-Vector) or treated with 2 μg/mL rAPOE3. Then, transwell assays were performed. Representative migration and invasion images are shown; (B) Quantitative results of transwell assays. The migrated and invaded cells were counted using ImageJ. APOE: Apolipoprotein E; rAPOE3: Recombinant APOE3; ns: No significance.

Article Snippet: Construction of APOE-overexpressing stable cell lines and cell culture Human colon cancer cell lines HCT116 and HCT8 (American Type Culture Collection, USA) were maintained in RPMI 1640 (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Gibco).

Techniques: Over Expression, Knockdown, Transfection, Plasmid Preparation, Migration, Recombinant

Journal: Biomolecules & biomedicine

Article Title: Jun-APOE-LRP1 axis promotes tumor metastasis in colorectal cancer.

doi: 10.17305/bb.2023.9248

Figure Lengend Snippet: Figure 5. LRP1 is highly expressed in CRC patients with lymphatic invasion and high APOE level, and APOE-overexpression upregulates LRP1 protein level. (A) The intracellular and extracellular apoE levels of HCT116 and HCT8 cells with APOE-overexpression were determined by ELISA; (B) Schematic diagram of protein interactions of apoE. Data were obtained from the STRING database; (C) Comparison of LRP1 mRNA levels between the lymphatic invasion group and the non-lymphatic invasion group; (D) Comparison of LRP1 mRNA levels between the APOEHigh group (median APOE mRNA level > 10.24) and APOELow group (median APOE mRNA level < 10.24). Data were obtained from the UCSC Xena database; (E) Relative LRP1 mRNA levels of CRC cells with or without APOE-overexpression; (F) Determination of LRP1 protein levels in HCT116 and HCT8 cells with or without APOE-overexpression using western blot. apoA1: Apolipoprotein A1; apoA2: Apolipoprotein A2; apoB: Apolipoprotein B; apoC1: Apolipoprotein C1; apoC2: Apolipoprotein C2; apoC3: Apolipoprotein C3; apoE: Apolipoprotein E; APP: Amyloid precursor protein; CRC: Colorectal cancer; LDLR: Low-density lipoprotein receptor; LRP1: Lipoprotein receptor-related protein 1; LRP8: Lipoprotein receptor-related protein 8; OE: Overexpression; VC: Vector control; ns: No significance; ELISA: Enzyme-linked immunosorbent assay.

Article Snippet: Construction of APOE-overexpressing stable cell lines and cell culture Human colon cancer cell lines HCT116 and HCT8 (American Type Culture Collection, USA) were maintained in RPMI 1640 (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Gibco).

Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Comparison, Western Blot, Plasmid Preparation, Control

Journal: Biomolecules & biomedicine

Article Title: Jun-APOE-LRP1 axis promotes tumor metastasis in colorectal cancer.

doi: 10.17305/bb.2023.9248

Figure Lengend Snippet: Figure 6. LRP1 mediates the metastasis-promoting function of APOE. HCT116 and HCT8 cells with APOE-overexpression were transfected with si-NC or si-LRP1, and the efficiency of LRP1 knockdown was validated by qPCR (A) and western blot (B); (C) Transwell assays were performed 24 h post-transfection, and representative images of migration and invasion were shown; (D) Quantitative results of transwell assays. The migrated and invaded cells were counted using ImageJ. APOE: Apolipoprotein E; LRP1: Lipoprotein receptor-related protein 1; OE: Overexpression; qPCR: Quantitative polymerase chain reaction; VC: Vector control; ns: No significance.

Article Snippet: Construction of APOE-overexpressing stable cell lines and cell culture Human colon cancer cell lines HCT116 and HCT8 (American Type Culture Collection, USA) were maintained in RPMI 1640 (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Gibco).

Techniques: Over Expression, Transfection, Knockdown, Western Blot, Migration, Real-time Polymerase Chain Reaction, Plasmid Preparation, Control

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: (A and B) LRP5 protein expression in SW480 (A) and LoVo (B) cells stably transduced with control (Ctrl) or 15-LOX-1 lentivirus and treated with 10 μg/mL cycloheximide (CHX) for 0, 2, and 4 h. The whole-cell lysates were analyzed using western blot.

Article Snippet: Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Expressing, Stable Transfection, Transduction, Western Blot

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: (A) SNX17, LRP5, and active β-catenin expression in 293T or SW480 cells transfected with control vector (Ctrl) or FLAG-tagged SNX17 expression vector for 48 h, then processed for western blot analysis.

Article Snippet: Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: (A and B) SW480 (A) or LoVo (B) cells stably transduced with control (Ctrl) or 15-LOX-1 lentivirus were treated with the PI3P biosynthesis inhibitor VPS34-IN-1 (1 μM) for 2 h and processed for the indicated protein expression using western blot analysis.

Article Snippet: Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Stable Transfection, Transduction, Expressing, Western Blot

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Generated, Plasmid Preparation, Recombinant, Modification, Western Blot, Protease Inhibitor, Cell Viability Assay, Magnetic Beads, Expressing, Software

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: (A and B) LRP5 protein expression in SW480 (A) and LoVo (B) cells stably transduced with control (Ctrl) or 15-LOX-1 lentivirus and treated with 10 μg/mL cycloheximide (CHX) for 0, 2, and 4 h. The whole-cell lysates were analyzed using western blot.

Article Snippet: EXPERIMENTAL MODEL AND SUBJECT DETAILS Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Expressing, Stable Transfection, Transduction, Western Blot

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: (A) SNX17, LRP5, and active β-catenin expression in 293T or SW480 cells transfected with control vector (Ctrl) or FLAG-tagged SNX17 expression vector for 48 h, then processed for western blot analysis.

Article Snippet: EXPERIMENTAL MODEL AND SUBJECT DETAILS Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: (A and B) SW480 (A) or LoVo (B) cells stably transduced with control (Ctrl) or 15-LOX-1 lentivirus were treated with the PI3P biosynthesis inhibitor VPS34-IN-1 (1 μM) for 2 h and processed for the indicated protein expression using western blot analysis.

Article Snippet: EXPERIMENTAL MODEL AND SUBJECT DETAILS Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Stable Transfection, Transduction, Expressing, Western Blot

Journal: Cell reports

Article Title: Suppression of Membranous LRP5 Recycling, Wnt/β-catenin Signaling, and Colon Carcinogenesis by 15-LOX-1 Peroxidation of Linoleic Acid in PI3P

doi: 10.1016/j.celrep.2020.108049

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: EXPERIMENTAL MODEL AND SUBJECT DETAILS Cell culture Human colon cancer cell lines SW480 and LoVo and human embryonic kidney cell line 293T were purchased from ATCC and were maintained in McCoy 5A, RPMI 1640, and Dulbecco modified Eagle (high-glucose) media, respectively.

Techniques: Generated, Plasmid Preparation, Recombinant, Modification, Western Blot, Protease Inhibitor, Cell Viability Assay, Magnetic Beads, Expressing, Software