hct15  (ATCC)

Journal: Medical Oncology (Northwood, London, England)

Article Title: Investigating the effects of platelets, platelet releasate and aspirin on colorectal cancer cell proliferation, migration and invasion

doi: 10.1007/s12032-026-03264-z

Figure Lengend Snippet: Platelet activation and binding to colorectal cancer cells. Whole blood obtained from healthy individuals ( n = 5) was treated with DPBS or 1mM aspirin before incubation with different concentrations of HCT15 or HCT116 colorectal cancer cells. Platelet activation was measured using flow cytometry for the platelet activation markers ( a ) P-selectin, ( b ) LAMP-3 and ( c ) activated GPIIb/IIIa. Representative images of platelets binding to d) HCT15 and e) HCT116 cells. Platelets were stained using an anti-CD41 antibody (green) and CRC cells were stained using an anti-EpCAM antibody (red) and DAPI (blue) and imaged using immunofluorescence microscopy. Data presented as box whisker plots for n = 5 experiments. * P < 0.05, ** P < 0.01

Article Snippet: HCT15 and HCT116 human colorectal cancer cells (ATCC, Manassas, USA), RPMI 1640 + L-glutamine, fetal bovine serum, penicillin/streptomycin and Dulbecco’s phosphate buffer saline (Gibco, Waltham, USA); Vacuette tube 9mL ACD-B (Greiner Bio-One, Kremsmünster, Austria); prostaglandin I 2 (PGI 2 ) sodium salt (Cayman Chemicals, Ann Arbor, USA); thrombin receptor activating peptide (Roche, Basel Switzerland); CD41 antibody (Abcam, Cambridge, UK); EpCAM (D4K8R) XP antibody, anti-mouse IgG (H + L), F(ab’)2 fragment Alexa Fluor 488, anti-rabbit IgG (H + L), F(ab’) 2 fragment Alexa Fluor 555 and ProLong Gold Antifade Reagent with DAPI (Cell Signalling Technology, Danvers, USA); Vacutainer Citrate 2.7mL tubes, TruCount tubes, CD42b PE, CD62p APC, CD63 PE-Cy7 and PAC-1 FITC antibodies (BD Biosciences, Franklin Lakes, USA); paraformaldehyde 16% solution, EM grade (Electron Microscopy Sciences, Hatfield, USA); sodium chloride 0.9% for irrigation (Baxter, Deerfield, USA); CellTrace Carboxyfluorescein succinimidyl ester (CFSE) Cell Proliferation Kit (Thermo Fisher Scientific, Waltham, USA); cell culture plates, Transwell 6.5 mm 8.0 μm polycarbonate insert, Matrigel and 5mL polystyrene round-bottom Tube 12 × 75 mm (Corning, Corning, USA), crystal violet solution, aspirin and phenazine methosulfate (PMS) (Sigma-Aldrich, St. Louis, USA).

Techniques: Activation Assay, Binding Assay, Incubation, Flow Cytometry, Staining, Immunofluorescence, Microscopy, Whisker Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: Investigating the effects of platelets, platelet releasate and aspirin on colorectal cancer cell proliferation, migration and invasion

doi: 10.1007/s12032-026-03264-z

Figure Lengend Snippet: The effect of platelets and platelet releasate on colorectal cancer cell migration and invasion. Inserts in transwell assays were ( a ) uncoated to measure migration or ( b ) Matrigel coated to measure invasion of HCT15 and HCT116 cells co-incubated with platelets or platelet-releasate from healthy individuals ( n = 5) for 24 h. Platelets were pretreated with DPBS or 1mM aspirin and platelet releasate was generated from platelets pretreated with DPBS or aspirin. Data is presented box whisker plots for n = 5 experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HCT15 and HCT116 human colorectal cancer cells (ATCC, Manassas, USA), RPMI 1640 + L-glutamine, fetal bovine serum, penicillin/streptomycin and Dulbecco’s phosphate buffer saline (Gibco, Waltham, USA); Vacuette tube 9mL ACD-B (Greiner Bio-One, Kremsmünster, Austria); prostaglandin I 2 (PGI 2 ) sodium salt (Cayman Chemicals, Ann Arbor, USA); thrombin receptor activating peptide (Roche, Basel Switzerland); CD41 antibody (Abcam, Cambridge, UK); EpCAM (D4K8R) XP antibody, anti-mouse IgG (H + L), F(ab’)2 fragment Alexa Fluor 488, anti-rabbit IgG (H + L), F(ab’) 2 fragment Alexa Fluor 555 and ProLong Gold Antifade Reagent with DAPI (Cell Signalling Technology, Danvers, USA); Vacutainer Citrate 2.7mL tubes, TruCount tubes, CD42b PE, CD62p APC, CD63 PE-Cy7 and PAC-1 FITC antibodies (BD Biosciences, Franklin Lakes, USA); paraformaldehyde 16% solution, EM grade (Electron Microscopy Sciences, Hatfield, USA); sodium chloride 0.9% for irrigation (Baxter, Deerfield, USA); CellTrace Carboxyfluorescein succinimidyl ester (CFSE) Cell Proliferation Kit (Thermo Fisher Scientific, Waltham, USA); cell culture plates, Transwell 6.5 mm 8.0 μm polycarbonate insert, Matrigel and 5mL polystyrene round-bottom Tube 12 × 75 mm (Corning, Corning, USA), crystal violet solution, aspirin and phenazine methosulfate (PMS) (Sigma-Aldrich, St. Louis, USA).

Techniques: Migration, Incubation, Generated, Whisker Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: Investigating the effects of platelets, platelet releasate and aspirin on colorectal cancer cell proliferation, migration and invasion

doi: 10.1007/s12032-026-03264-z

Figure Lengend Snippet: The effect of platelets and platelet releasate on colorectal cancer cell proliferation. a ) HCT15 and b ) HCT116 cells stained with CFSE were co-incubated with platelets or platelet releasate from healthy individuals ( n = 6) for 48 h. Platelets were pretreated with DPBS or 1mM aspirin and platelet releasate was generated from platelets pretreated with DPBS or aspirin. The cells were analysed using flow cytometry and ModFit LT software which calculated a proliferation index. Data is presented box whisker plots for n = 6 experiments. ( c ) HCT15 and ( d ) HCT116 cells were co-incubated for 48 h with platelets from healthy individuals ( n = 5) pretreated with DPBS or 1mM aspirin, with proliferation determined using MTS. Data is presented as the mean with error bars representing the standard error of the mean for n = 5 experiments. ( e ) HCT15 and ( g ) HCT116 cells were incubated with 1mM aspirin for 48 h ( n = 10) and the effect it had on proliferation was determined using MTS proliferation assays. Data is presented box whisker plots for n = 10 experiments. ** P < 0.01

Article Snippet: HCT15 and HCT116 human colorectal cancer cells (ATCC, Manassas, USA), RPMI 1640 + L-glutamine, fetal bovine serum, penicillin/streptomycin and Dulbecco’s phosphate buffer saline (Gibco, Waltham, USA); Vacuette tube 9mL ACD-B (Greiner Bio-One, Kremsmünster, Austria); prostaglandin I 2 (PGI 2 ) sodium salt (Cayman Chemicals, Ann Arbor, USA); thrombin receptor activating peptide (Roche, Basel Switzerland); CD41 antibody (Abcam, Cambridge, UK); EpCAM (D4K8R) XP antibody, anti-mouse IgG (H + L), F(ab’)2 fragment Alexa Fluor 488, anti-rabbit IgG (H + L), F(ab’) 2 fragment Alexa Fluor 555 and ProLong Gold Antifade Reagent with DAPI (Cell Signalling Technology, Danvers, USA); Vacutainer Citrate 2.7mL tubes, TruCount tubes, CD42b PE, CD62p APC, CD63 PE-Cy7 and PAC-1 FITC antibodies (BD Biosciences, Franklin Lakes, USA); paraformaldehyde 16% solution, EM grade (Electron Microscopy Sciences, Hatfield, USA); sodium chloride 0.9% for irrigation (Baxter, Deerfield, USA); CellTrace Carboxyfluorescein succinimidyl ester (CFSE) Cell Proliferation Kit (Thermo Fisher Scientific, Waltham, USA); cell culture plates, Transwell 6.5 mm 8.0 μm polycarbonate insert, Matrigel and 5mL polystyrene round-bottom Tube 12 × 75 mm (Corning, Corning, USA), crystal violet solution, aspirin and phenazine methosulfate (PMS) (Sigma-Aldrich, St. Louis, USA).

Techniques: Staining, Incubation, Generated, Flow Cytometry, Software, Whisker Assay

Journal: Frontiers in Immunology

Article Title: Unveiling tumor senescence-driven prognostic heterogeneity via MALISS in stage II/III colorectal cancer

doi: 10.3389/fimmu.2025.1744719

Figure Lengend Snippet: NR1D2 promotes the CRC migration. (A) The importance of 30 immunosenescence-related genes selected by Coxboost algorithm. (B) Kaplan-Meier survival curve of PFS between patients with high NR1D2 and low NR1D2 mRNA level. (C) The NR1D2 expression of mRNA and protein levels within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3. Relative band intensity (normalized) is shown below the WB image. (D) SA-β-gal staining to analyze the senescence status in the HCT15 cells within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3. (E) P16 and P21 protein levels assessed by Western blot after NR1D2 knocking down. (F) Changes in mRNA expression of SASP-associated secretory factors (TGF-β, GDF15) as determined by qRT-PCR. (G) Wound healing assays was performed to detect the migration within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3 in HCT15. (H) Transwell assays was performed to detect the migration within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3 in HCT15. * means P < 0.05, ** means P < 0.01, *** means P < 0.001, **** means P < 0.0001.

Article Snippet: The HCT15 cell line (RRID: CVCL_0292), which was cultured in RPMI-1640 (Gibco) medium with 10% fetal bovine serum (Vazyme), penicillin-streptomycin (100 U/mL, NCM), was procured from ATCC.

Techniques: Migration, Expressing, Control, Staining, Western Blot, Quantitative RT-PCR