human colorectal cancer cell lines  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Network Pharmacology and Molecular Docking-Based Approach Revealing the Potential Anticancer Compounds and Molecular Mechanisms of Paris polyphylla Against Colorectal Cancer

doi: 10.3390/ijms27093874

Figure Lengend Snippet: Cytotoxic effects of Paris polyphylla rhizome extract (PPRE) on colorectal cancer cells. SW480 ( A ) and HCT116 ( B ) cells were treated with increasing concentrations of PPRE (0–80 µg/mL) for 24 and 48 h. Data are presented as mean ± SD from three independent experiments. Statistical significance compared with the untreated control group was determined using one-way ANOVA followed by Tukey’s multiple comparison test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Human colorectal cancer cell lines SW480 and HCT116 (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Network Pharmacology and Molecular Docking-Based Approach Revealing the Potential Anticancer Compounds and Molecular Mechanisms of Paris polyphylla Against Colorectal Cancer

doi: 10.3390/ijms27093874

Figure Lengend Snippet: qRT-PCR Validation of STAT3, EGFR, SRC, IL-6, and AKT1 mRNA expression in SW480 ( A – E ) and HCT116 ( F – J ) cells following treatment with PPRE (0–10 µg/mL). Gene expression levels were normalized to the internal control and expressed as fold change relative to untreated control (0 µg/mL). Data are presented as mean ± SD of three independent experiments. Statistical significance versus the untreated control was determined using one-way ANOVA followed by Tukey’s post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Human colorectal cancer cell lines SW480 and HCT116 (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Quantitative RT-PCR, Biomarker Discovery, Expressing, Gene Expression, Control

Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Venn diagram showing overlapping differentially expressed genes (DEGs) between GSE44076 , GSE9348 , GSE23878 datasets and a curated ferroptosis-related gene set. B Heatmap demonstrating expression patterns of four overlapping genes in CRC versus normal tissues from the TCGA database. C , D CCK-8 assays measuring viability of SW480 cells after siRNA-mediated knockdown of AURKA, SCD, ETV4, and DPEP1 ( C ) and following RSL3 treatment for 48 h ( D ). E , F Western blot analysis of AURKA protein levels across various human CRC cell lines ( E ) or validation of shRNA-mediated stable AURKA knockdown efficiency in SW480 and RKO cells ( F ). G IC 50 analysis of RSL3 in CRC (shNC and sh AURKA ) cells treated with different concentrations of RSL3 for 48 h. H Representative TEM images of SW480 (shNC and sh AURKA ) cells after the indicated treatment with RSL3 (5 μM) for 24 h. I Representative FACS images and quantified values of lipid ROS levels in SW480 (shNC and sh AURKA ) cells treated with RSL3 (5 μM) for 24 h. J , K Intracellular GSH/GSSG ratios ( J ) and cellular concentrations of MDA ( K ) were evaluated in SW480 (shNC and sh AURKA ) cells. L AURKA mRNA levels in normal versus tumor specimens from TCGA colon adenocarcinoma (COAD) and rectum adenocarcinoma (READ) database. M Western blot analysis of AURKA protein expression in 12 paired adjacent normal (N) and CRC (T) tissues from SYSU-FAH cohort, with quantitative normalization to β-actin. N qPCR analysis of AURKA mRNA expression in 22 paired CRC tissues from SYSU-FAH cohort. O IHC staining and quantitative analysis of AURKA expression in CRC tumor microarrays (TMAs). Scale bars: 50 µm. P Overall survival analysis based on AURKA expression in Kaplan-Meier Plotter CRC cohort ( n = 1061). Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test [( I ), ( J ), ( K ), ( L ), and ( O )], paired Student’s t test [( M ) and ( N )], two-way ANOVA test ( C ), or log-rank test ( P ). *** p < 0.001.

Article Snippet: The human colorectal cancer cell lines SW480 (RRID: CVCL_0546), RKO (RRID: CVCL_0504), HCT116 (RRID: CVCL_0291), DLD1 (RRID: CVCL_0248), SW1116 (RRID: CVCL_1724), SW620 (RRID: CVCL_0547) and human embryonic kidney (HEK) 293 T (RRID: CVCL_0063) were obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, CCK-8 Assay, Knockdown, Western Blot, Biomarker Discovery, shRNA, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Western blot analysis of SLC7A11, GPX4, and ACSL4 in SW480 (shNC and sh AURKA ) cells. B , C RNA-seq with KEGG enrichment analysis in SW480 (shNC and sh AURKA ) cells showing upregulated pathways in sh AURKA group ( B ), and a heatmap depicting DEGs in sterol/cholesterol biosynthesis pathways ( C ). D Schematic of the cholesterol biosynthesis pathway and its regulatory enzymes. E qPCR quantification of cholesterol biosynthesis enzyme mRNA levels in CRC (shNC and sh AURKA ) cells. F , G Western blot analysis of DHCR7 protein expression in SW480 and RKO cells ( F ), and SW480 cells following siRNA knockdown ( G ). H IC 50 analysis of RSL3 in SW480 (shNC and sh AURKA ) cells treated with RSL3 (48 h) following DHCR7 silencing. I Representative FACS images and quantified values of lipid ROS levels in SW480 (shNC and sh AURKA ) cells treated with RSL3 (5 μM, 24 h) following DHCR7 silencing. J LC-MS/MS quantification of 7-DHC levels in CRC (shNC and sh AURKA ) cells. K Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with 7-DHC (25 μM), cholesterol (25 μM) for 24 h. L Diagram of cholesterol synthesis showing DHCR7 converting 7-DHC to cholesterol and its inhibition by AY9944. M Intracellular levels of 7-DHC in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. N Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with AY9944 (100 nM) for 24 h. O, P Quantification of intracellular lipid ROS ( O ) and MDA ( P ) levels in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test [( E ), ( I ), ( J ), ( M , O , P )]. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The human colorectal cancer cell lines SW480 (RRID: CVCL_0546), RKO (RRID: CVCL_0504), HCT116 (RRID: CVCL_0291), DLD1 (RRID: CVCL_0248), SW1116 (RRID: CVCL_1724), SW620 (RRID: CVCL_0547) and human embryonic kidney (HEK) 293 T (RRID: CVCL_0063) were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, RNA Sequencing, Expressing, Knockdown, Liquid Chromatography with Mass Spectroscopy, Inhibition

Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Venn diagram analysis of transcription factors predicated to regulate DHCR7 gene transcription based on ChIP-Atlas, GTRD, and hTFtarget databases. B Western blot analysis of SREBP2, c-Myc, ZEB1, and MAX in CRC (shNC and sh AURKA ) cells. C Western blot analysis of SREBP2 and DHCR7 protein levels in SW480 cells after siRNA-mediated SREBP2 knockdown. D qPCR quantification of cholesterol biosynthesis enzyme mRNA levels in SREBP2-silenced SW480 cells. E IC 50 analysis of RSL3 in SREBP2-silenced SW480 (shNC and sh AURKA ) cells treated with RSL3 (48 h). F , G Representative FACS images and quantified values of lipid ROS levels ( F ) and GSH/GSSG ratios ( G ) in SREBP2-silenced SW480 (shNC and sh AURKA ) cells treated with RSL3 (5 μM, 24 h). H , I Western blot analysis of SREBP2 at indicated timepoints after CHX (100 µg/ml) treatment ( H ), and quantified degradation kinetics ( I ) in SW480 cells. J Immunofluorescence (IF) analyses of subcellular distribution of SREBP2 in SW480 (shNC and sh AURKA ) cells. K , L Western blot analysis of pSREBP2, nSREBP2, total AMPK, and pAMPK levels in SW480 (shNC and sh AURKA ) cells ( K ) and post AICAR treatment (1 mM, 24 h, L ). M IC 50 analysis of RSL3 in SW480 (shNC and sh AURKA ) cells after pretreatment with AICAR (1 mM, 24 h). N Representative FACS images and quantified values of lipid ROS levels in SW480 (shNC and sh AURKA ) cells treated with AICAR (1 mM, 24 h). Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test [( D ), ( F ), ( G ), and ( N )]. ** p < 0.01, *** p < 0.001.

Article Snippet: The human colorectal cancer cell lines SW480 (RRID: CVCL_0546), RKO (RRID: CVCL_0504), HCT116 (RRID: CVCL_0291), DLD1 (RRID: CVCL_0248), SW1116 (RRID: CVCL_1724), SW620 (RRID: CVCL_0547) and human embryonic kidney (HEK) 293 T (RRID: CVCL_0063) were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Knockdown, Immunofluorescence

Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Immunoprecipitation-mass spectrometry (IP-MS) analysis of AURKA-interacting kinases/phosphatases. B Top 10 AURKA-binding proteins identified by mass spectrometry. C Representative SAPS3 peptide spectrum from IP-MS. D Co-IP assay showing the interaction between exogenous Flag-AURKA with HA-SAPS3 in SW480 and RKO cells. E Endogenous AURKA-SAPS3 interaction confirmed by Co-IP using anti-AURKA antibody in SW480 and RKO cells. F Immunofluorescent staining showing co-localization of endogenous AURKA and SAPS3 in SW480 and RKO cells. Scale bars: 5 µm. G Western blot analysis of SAPS3 in SW480 and RKO cells. H , I Western blot analysis of SAPS3 at indicated timepoints after CHX (100 µg/ml) treatment ( H ), and quantified degradation kinetics ( I ) in SW480 cells. J IC 50 analysis of RSL3 in SW480 (shNC and sh AURKA ) cells treated with RSL3 (48 h) following SAPS3 knockdown. K Representative FACS images and quantified values of lipid ROS levels in SW480 (shNC and sh AURKA ) cells after SAPS3 silencing. L Western blot analysis of total AMPK, pAMPK, pSREBP2, nSREBP2, and DHCR7 levels in SW480 (shNC and sh AURKA ) cells post SAPS3 knockdown. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test ( K ) and two-way ANOVA test ( I ). *** p < 0.001.

Article Snippet: The human colorectal cancer cell lines SW480 (RRID: CVCL_0546), RKO (RRID: CVCL_0504), HCT116 (RRID: CVCL_0291), DLD1 (RRID: CVCL_0248), SW1116 (RRID: CVCL_1724), SW620 (RRID: CVCL_0547) and human embryonic kidney (HEK) 293 T (RRID: CVCL_0063) were obtained from the American Type Culture Collection (ATCC).

Techniques: Immunoprecipitation, Mass Spectrometry, Protein-Protein interactions, Binding Assay, Co-Immunoprecipitation Assay, Staining, Western Blot, Knockdown

Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Co-IP analysis of SAPS3 indicating reduced phosphorylation levels in SW480-sh AURKA cells. B Co-IP analysis of the phosphorylation levels of SAPS3 in SW480-sh AURKA cells reconstituted with WT-, T288D-, or D274N-AURKA. C Western blot analysis of SAPS3, total AMPK, pAMPK, pSREBP2, nSREBP2, and DHCR7 in SW480-sh AURKA cells expressing WT-, T288D-, or D274N-AURKA. D IC 50 analysis of RSL3 in SW480-sh AURKA cells expressing WT-, T288D-, or D274N-AURKA. E Representative FACS images and quantified values of lipid ROS levels in SW480-sh AURKA cells expressing WT-, T288D-, or D274N-AURKA. F LC-MS/MS identified Ser524 as AURKA-dependent phosphorylation sites in SAPS3. G Evolutionary conservation of SAPS3 S523/S524/Ser525 phosphorylation motifs across species (sequence alignment). H , I Co-IP analysis of phosphorylation levels for WT-, S523A-, S524A-, and S525A-SAPS3 mutants ( H ) or D-mut SAPS3 ( I ) in SW480 cells. J Co-IP analysis of WT or D-mut SAPS3 phosphorylation in SW480 cells treated with/without Alisertib (5 µM). K Western blot analysis of total AMPK, pAMPK, pSREBP2, nSREBP2, and DHCR7 levels in SW480 cells transfected with WT- or D-mut SAPS3, with/without Alisertib treatment. L , N Western blot analysis of WT- ( L ) or D-mut SAPS3 ( N ) protein stability at indicated timepoints after CHX (100 µg/ml) treatment in SW480-sh AURKA cells, comparing AURKA-reconstituted versus control conditions. M , O Quantified degradation kinetics (half-life) of WT- ( M ) or D-mut SAPS3 ( O ) from ( L , N ), respectively. P Western blot analysis of WT- or D-mut SAPS3 stability in CHX-treated SW480 cells, with/without MG132 (10 µM) under AURKA-expressing conditions. Q Western blot analysis of WT- or D-mut SAPS3 stability in CHX-treated SW480 cells, with/without BafA1 (200 ng/ml) under AURKA-expressing conditions. R Ubiquitination assays of WT- or D-mut SAPS3 in SW480 cells with/without AURKA expression. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test ( E ) and two-way ANOVA test ( M , O ). *** p < 0.001.

Article Snippet: The human colorectal cancer cell lines SW480 (RRID: CVCL_0546), RKO (RRID: CVCL_0504), HCT116 (RRID: CVCL_0291), DLD1 (RRID: CVCL_0248), SW1116 (RRID: CVCL_1724), SW620 (RRID: CVCL_0547) and human embryonic kidney (HEK) 293 T (RRID: CVCL_0063) were obtained from the American Type Culture Collection (ATCC).

Techniques: Co-Immunoprecipitation Assay, Phospho-proteomics, Western Blot, Expressing, Liquid Chromatography with Mass Spectroscopy, Sequencing, Transfection, Control, Ubiquitin Proteomics

Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Dose-response analysis of 5-FU sensitivity in CRC (shNC and sh AURKA ) cells treated for 48 h. B Representative images (left) and quantification (right) of clonogenic assay with 500 SW480 cells/well treated with 5-FU (60 μM). C , D Tumor growth curves ( C ) and representative images of excised tumors with their weights at endpoint ( D ) in SW480 (shNC and sh AURKA ) implanted nude mice, followed by i.p . injection of 5-FU (25 mg/kg) or PBS ( n = 5 mice per group). E Viability of SW480 (shNC and sh AURKA ) cells with or without 5-FU (60 μM) for 48 h in combination with Necrostatin (Nec, 20 μM), Chloroquine (CQ, 30 μM), Disulfiram (Dis, 1 μM), Z-VAD-FMK (VAD, 25 μM) and Ferrostatin-1 (Fer, 10 μM). F Representative TEM images of SW480 (shNC and sh AURKA ) cells after 5-FU (60 μM) treatment for 24 h. G , H Representative FACS images and quantified lipid ROS levels ( G ) and GSH/GSSG ratios ( H ) in SW480 (shNC and sh AURKA ) cells treated with 5-FU (60 μM, 24 h). I , J Representative H&E and IHC staining images ( I ) and quantification ( J ) of Ki67, 4-HNE, Bax, and Bcl-2 staining in xenograft tumor sections. K Western blot analysis of AURKA, SAPS3, total AMPK, pAMPK, pSREBP2, nSREBP2, and DHCR7 in SW480 xenograft tumors with or without 5-FU treatment. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test ( B , D , E , G , H , J ) and two-way ANOVA test ( C ). ** p < 0.01, *** p < 0.001.

Article Snippet: The human colorectal cancer cell lines SW480 (RRID: CVCL_0546), RKO (RRID: CVCL_0504), HCT116 (RRID: CVCL_0291), DLD1 (RRID: CVCL_0248), SW1116 (RRID: CVCL_1724), SW620 (RRID: CVCL_0547) and human embryonic kidney (HEK) 293 T (RRID: CVCL_0063) were obtained from the American Type Culture Collection (ATCC).

Techniques: Clonogenic Assay, Injection, Immunohistochemistry, Staining, Western Blot

Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Western blot analysis of pAURKA in SW480 cells treated with Alisertib. B RSL3-IC 50 analysis in SW480 cells after Alisertib treatment (1 µM & 5 µM). C - E Representative FACS plots and quantified lipid ROS levels ( C ), MDA levels ( D ), and GSH/GSSG ratios ( E ) in SW480 cells treated with RSL3 (5 µM) and Alisertib. F Western blot analysis of AURKA, pAURKA, SAPS3, AMPK, pAMPK, pSREBP2, nSREBP2, and DHCR7 in Alisertib-treated SW480 cells. G LC-MS/MS quantification of 7-DHC and cholesterol levels in SW480 cells post Alisertib treatment. H Clonogenic assay showing synergistic suppression of SW480 survival with Alisertib and 5-FU (Combenefit synergy analysis). I , K Excised tumor images ( I ), tumor growth curves ( J ) and endpoint tumor weights ( K ) from PDX model after 5-FU (25 mg/kg, i.p.) and/or Alisertib (30 mg/kg, oral gavage) (n = 5 mice per group). L, M Representative H&E and IHC staining images ( L ) and quantification ( M ) of Ki67 and 4-HNE staining in PDX tumor sections. N Western blot analysis of AURKA, pAURKA, SAPS3, AMPK, pAMPK, pSREBP2, nSREBP2, and DHCR7 in Alisertib and/or 5-FU-treated PDX tumors. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test ( C , D , E , G , K , M ) and two-way ANOVA test ( J ). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The human colorectal cancer cell lines SW480 (RRID: CVCL_0546), RKO (RRID: CVCL_0504), HCT116 (RRID: CVCL_0291), DLD1 (RRID: CVCL_0248), SW1116 (RRID: CVCL_1724), SW620 (RRID: CVCL_0547) and human embryonic kidney (HEK) 293 T (RRID: CVCL_0063) were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Liquid Chromatography with Mass Spectroscopy, Clonogenic Assay, Immunohistochemistry, Staining