dld1  (ATCC)

Journal: Cell Reports Medicine

Article Title: Engineered oncolytic virus armed with anti-PCSK9 scFv boosts long-term CD8 + T cell immunity via rewiring MHC-I antigen presentation

doi: 10.1016/j.xcrm.2026.102724

Figure Lengend Snippet: OVs impaired CD8 + T cell response via inducing MHC-I degradation (A) C57 mice with MC38 subcutaneous xenografts were treated with VSVΔ51 (2 × 10 7 plaque-forming unit (PFU)/day, i.t., intra-tumoral injection). The CD45 + cells were isolated from MC38 tumor at 16 days for scRNA-seq analysis ( GSE293436 ). (B) UMAP plot of the 10 major cell types in MC38 tumor-bearing mice. (C) KEGG analysis demonstrates the changed signaling in T cell cluster following VSVΔ51 treatment. (D and E) LDH release assay was performed to detect the cytotoxicity of T lymphocytes. CT26-OVA cells were co-cultured with the OT1 T cells at indicated ratio (E:T = 1:1, 1:5, or 1:10) in the CM-Uninfected, CM-VSVΔ51-UV(D), or CM-VSVΔ51ΔG (E) for 24 h. (F and G) Flow cytometry analysis and quantification of cell surface MHC-I expression. CT26 cells were treated with CM-Uninfected, CM-VSVΔ51-UV (F), or CM-VSVΔ51ΔG (G) for 24 h. Then the expression of MHC-I was measured by flow cytometry. (H) DLD1 and HCT116 were infected with escalating titers of VSVΔ51 for 24 h. Or DLD1 and HCT116 were infected with VSVΔ51, MOI = 0.05, for different times. Flow cytometry plots and quantification of cell-surface MHC-I. Statistical significance was determined using two-tailed unpaired Student’s t test in (D, E, F, and G) and one-way ANOVA in (H). Data represent the mean ± SD. n = 3 biological replicates in (D, E, F, G, and H). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, nonsignificant.

Article Snippet: DLD1 , Procell , CL-0074.

Techniques: Injection, Isolation, Lactate Dehydrogenase Assay, Cell Culture, Flow Cytometry, Expressing, Infection, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Engineered oncolytic virus armed with anti-PCSK9 scFv boosts long-term CD8 + T cell immunity via rewiring MHC-I antigen presentation

doi: 10.1016/j.xcrm.2026.102724

Figure Lengend Snippet: OV infection induces the SREBP2-PCSK9 axis to degrade MHC-I through the lysosomal pathway (A) GSEA analysis showed an enrichment of gene signatures associated with the regulation of cholesterol metabolic process in DLD1 infected with VSVΔ51, MOI = 0.5, for 16 h ( n = 3). (B) qPCR analysis for the mRNA level of SREBP2 and PCSK9 genes. DLD1 and HCT116 cells infected with or without VSVΔ51 (MOI = 0.1, 16 h). (C) Immunoblotting analysis of the expression of VSVG, MHC-I, PCSK9, SREBP2, and GAPDH. HCT116 were infected with indicated titers of VSVΔ51 for 24 h. (D) ELISA assay for detecting the secretion of PCSK9. DLD1 and HCT116 were infected with VSVΔ51 (MOI = 0.5) for 24 h. (E and F) The anti-PCSK9 antibody (alirocumab) rescued the MHC-I downregulation induced by VSVΔ51 infection. The protein levels of MHC-I in DLD1 cells treated with VSVΔ51 (MOI = 0.05, 24 h) and alirocumab (400 μg/mL, 36 h) were detected by flow cytometry analysis (F) or immunoblotting (E). (G and H) PCSK9 knockout reversed the MHC-I downregulation induced by VSVΔ51 infection. The knockout efficiency of PCSK9 was verified by immunoblotting in HCT116 cells (G). The flow cytometry analysis of MHC-I in sgNC cells or sgPCSK9 cells treated with VSVΔ51 (MOI = 0.05, 24 h) or not (H). (I–K) The chemical or genomic inhibition of SREBP2 reversed the MHC-I downregulation induced by VSVΔ51 infection. Flow cytometry analysis of MHC-I treated with VSVΔ51 (MOI = 0.05, 24 h) and SREBP2 inhibitor fatostain (10 μM, 24 h) in DLD1 cells (I). Immunoblotting analysis for verifying the knockdown efficiency of SREBF2 in DLD1 cells (J). Flow cytometry of MHC-I in shNC cells or shSREBF2 DLD1 cells treated with VSVΔ51 (MOI = 0.05, 24 h) or not (K). (L) GSEA analysis of RNA-seq data showed an enrichment of gene signatures associated with processing and presentation of peptide antigen via MHC class I in VSVΔ51-treated DLD1 cells. DLD1 was infected with VSVΔ51, MOI = 0.5, for 16 h ( n = 3). (M) qPCR analysis of the MHC-I pathway genes in DLD1 cells infected with or without VSVΔ51 (MOI = 0.1, 16 h). (N) Immunoblotting analysis of MHC-I expression in DLD1 cells treated with cycloheximide and VSVΔ51 (MOI = 0.05, 24 h) or not. (O) Flow cytometry analysis of cell-surface MHC-I. DLD1 cells were infected with VSVΔ51 (MOI = 0.05, 24 h); then cells were treated with DMSO, MG132 (10 μM) for 8 h, or Baf A1 (autophagy inhibitor, 100 nM) for 16 h. Statistical significance was determined using two-tailed unpaired Student’s t test in (D, E, F and G) and one-way ANOVA in (H). Data represent the mean ± SD. n = 3 biological replicates in (D, E, F, G, and H). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, nonsignificant.

Article Snippet: DLD1 , Procell , CL-0074.

Techniques: Infection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Knock-Out, Inhibition, Knockdown, RNA Sequencing, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Engineered oncolytic virus armed with anti-PCSK9 scFv boosts long-term CD8 + T cell immunity via rewiring MHC-I antigen presentation

doi: 10.1016/j.xcrm.2026.102724

Figure Lengend Snippet: Engineered VSV carrying anti-PCSK9 scFv exhibited higher viral replication and tumor tropism (A) Schematic representation of rVSV-Ali or rVSV-Evo engineering: the target gene (alirocumab/evolocumab scFv-His) was integrated between the genes encoding the viral glycoprotein and large protein. (B and C) The construction of engineered VSV carrying anti-PCSK9 scFv (rVSV-Ali or rVSV-Evo). DLD1 cells were infected with VSVΔ51, rVSV-Ali, or rVSV-Evo (MOI = 0.5) for 24 h. The expression efficiency of anti-PCSK9 scFv in the infected cells was analyzed by qPCR (B) and immunoblot analysis (C). (D and E) The functional verification of rVSV-Ali and rVSV-Evo. DLD1 and HCT116 cells were infected with VSVΔ51, rVSV-Ali, or rVSV-Evo (MOI = 0.5, 16 h) and then were analyzed by flow cytometry for MHC-I expression (D). Tumor cells were treated with CM from infected cells (UV inactivation for 30 min) for 24 h. Then the surface MHC-I expression was measured by flow cytometry (E). (F) Plaque assay was conducted to detect the viral replication in DLD1 and HCT116 cells. The representative images of plaques were presented, and the viral titers were calculated. (G) The oncolytic effect of VSVΔ51, rVSV-Ali, and rVSV-Evo in DLD1 (MOI = 0.01) and HCT116 (MOI = 0.001) was detected by CCK-8 assay. (H–J) The in vivo analyses of tumor tropism and viral replication ( n = 5). The BALB/c mice bearing CT26 subcutaneous tumor were treated with VSVΔ51, rVSV-Ali, or rVSV-Evo (3 × 10 7 PFU, intravenous injection, qd×2). The mRNA of viral gene VSV-G was measured by qPCR in different tissues (H). Immunoblot analysis of VSV-G, MHC-I, PCSK9, SREBP2, His tag, and G4S linker in tumor tissue (I) and VSVG, MHC-I, PCSK9, LDLR, His tag, and G4S linker in liver (J). (K) The in vivo analyses of safety ( n = 5). Treatment regimens: qd, once daily; q2d, every two days. The BALB/c mice bearing CT26 subcutaneous tumor were treated with VSVΔ51 + alirocumab (3 × 10 7 PFU, intravenous injection, qd×2; 10 mg/kg, intraperitoneal injection, q2d×5) or rVSV-Ali (3 × 10 7 PFU, intravenous injection, qd×2). The total and free cholesterol levels were measured in serum, liver, and tumor. Statistical significance was determined using two-tailed unpaired Student’s t test in (B and G) and one-way ANOVA in (D, E, F, H, and K). Data represent the mean ± SD. n = 3 biological replicates in (B, D, E, F, G, and I). n = 5 biological replicates in (H and K). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, nonsignificant.

Article Snippet: DLD1 , Procell , CL-0074.

Techniques: Infection, Expressing, Western Blot, Functional Assay, Flow Cytometry, Plaque Assay, CCK-8 Assay, In Vivo, Injection, Two Tailed Test

Journal: Plant Foods for Human Nutrition (Dordrecht, Netherlands)

Article Title: Gastrointestinal Digestion Enhances the In vitro Antitumor Activity of Red Grape Anthocyanins Against Colorectal Cancer Cells

doi: 10.1007/s11130-026-01498-w

Figure Lengend Snippet: In vitro antitumor activity of undigested and digested red grape anthocyanin extracts in DLD1 and Caco2 cells. ( A ) MTT cell viability curves; ( B ) IC₅₀ values and relative potency; ( C ) Representative cell morphology of untreated (CTR) and IC₅₀-treated cells; ( D ) Trypan Blue viability validation in DLD1 cells. Statistical significance was assessed by t-test and Extra Sum-of-Squares F Test

Article Snippet: Two human colorectal carcinoma cell lines, Caco2 (ATCC, USA) and DLD1 (ECACC), were cultured under standard conditions as detailed in SM.

Techniques: In Vitro, Activity Assay, Biomarker Discovery

Journal: Plant Foods for Human Nutrition (Dordrecht, Netherlands)

Article Title: Gastrointestinal Digestion Enhances the In vitro Antitumor Activity of Red Grape Anthocyanins Against Colorectal Cancer Cells

doi: 10.1007/s11130-026-01498-w

Figure Lengend Snippet: Transcriptomic alterations in DLD1 cells treated with IC₅₀ of digested red grape anthocyanins. ( A ) Hierarchical clustering of microarray samples; ( B ) Volcano plot of differentially expressed genes (|FR| ≥ 1.5; adj. p < 0.05); ( C ) Heatmap of top 50 up- and downregulated genes; ( D ) Enriched Hallmark and Reactome gene sets identified by GSEA (FDR < 25%, nominal p < 0.05)

Article Snippet: Two human colorectal carcinoma cell lines, Caco2 (ATCC, USA) and DLD1 (ECACC), were cultured under standard conditions as detailed in SM.

Techniques: Microarray

Journal: Plant Foods for Human Nutrition (Dordrecht, Netherlands)

Article Title: Gastrointestinal Digestion Enhances the In vitro Antitumor Activity of Red Grape Anthocyanins Against Colorectal Cancer Cells

doi: 10.1007/s11130-026-01498-w

Figure Lengend Snippet: Apoptosis-related effects of digested red grape anthocyanins in DLD1 cells. ( A ) Enriched apoptosis-related gene sets (GSEA); ( B ) Annexin V/PI apoptosis quantification at IC₅₀ and 5 µg/mL; (C) RT-qPCR validation of selected apoptosis-associated genes. * p < 0.05, ** p < 0.01, *** p < 0.001 (t-test)

Article Snippet: Two human colorectal carcinoma cell lines, Caco2 (ATCC, USA) and DLD1 (ECACC), were cultured under standard conditions as detailed in SM.

Techniques: Quantitative RT-PCR, Biomarker Discovery

Journal: Plant Foods for Human Nutrition (Dordrecht, Netherlands)

Article Title: Gastrointestinal Digestion Enhances the In vitro Antitumor Activity of Red Grape Anthocyanins Against Colorectal Cancer Cells

doi: 10.1007/s11130-026-01498-w

Figure Lengend Snippet: Proliferation-related effects of digested red grape anthocyanins in DLD1 cells. ( A ) AlamarBlue proliferation kinetics at IC₅₀ and 5 µg/mL; ( B ) Cell cycle phase distribution by flow cytometry; ( C ) RT-qPCR validation of selected proliferation-associated genes. * p < 0.05, ** p < 0.01, *** p < 0.001 (t-test)

Article Snippet: Two human colorectal carcinoma cell lines, Caco2 (ATCC, USA) and DLD1 (ECACC), were cultured under standard conditions as detailed in SM.

Techniques: Flow Cytometry, Quantitative RT-PCR, Biomarker Discovery

Journal: Plant Foods for Human Nutrition (Dordrecht, Netherlands)

Article Title: Gastrointestinal Digestion Enhances the In vitro Antitumor Activity of Red Grape Anthocyanins Against Colorectal Cancer Cells

doi: 10.1007/s11130-026-01498-w

Figure Lengend Snippet: Cell motility-related effects of digested red grape anthocyanins in DLD1 cells. ( A ) Enriched motility-related Reactome gene sets (GSEA); ( B ) Spatial characteristics of the microfluidic device; ( C ) Confined 3D migration parameters (speed, velocity, migratory cell number) at IC₅₀ and 5 µg/mL. * p < 0.05, ** p < 0.01, *** p < 0.001 (t-test)

Article Snippet: Two human colorectal carcinoma cell lines, Caco2 (ATCC, USA) and DLD1 (ECACC), were cultured under standard conditions as detailed in SM.

Techniques: Migration