human pancreatic cancer cell lines panc 1  (ATCC)

Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Agrimol B inhibits PDAC growth both in vitro and in vivo . (A) A CCK-8 assay was used to evaluate the pharmacodynamic patterns of 62 natural polyphenols. (B) Polyphenol classification and chemical structure. (C) Viability of PANC-1 cells treated with 62 drugs (10 μmol/l). (D) Chemical structure of Agrimol B. (E) PANC-1, CFPAC-1, and AsPC-1 cell lines were treated with various concentrations of Agrimol B for 24 h. (F) IC50 of Agrimol B in PDAC cells. (G, H) PDAC cells were treated with the indicated concentrations of Agrimol B. Cell proliferation was measured by a colony formation assay. (I) Flow cytometric analysis of PDAC cells treated with different concentrations of Agrimol B for 24 h. (J) Western blot analysis of caspase 9 and cleaved caspase 9 in PDAC cells treated with the indicated concentrations of Agrimol B for 24 h. (K) Tumours from the xenograft model treated with Agrimol B or vehicle. (M, N) Tumour volume and weight in mice in the cohorts treated with Agrimol B or vehicle. (L, O) Immunohistochemical staining of Ki67 in a xenograft model from the vehicle group or the Agrimol B group. Relative immunohistochemical scores were determined by multiplying the range of positively stained cells. Scale bars, 100 μm.

Article Snippet: First, we performed an unbiased, comparative analysis of therapeutic responses among 62 polyphenolic compounds (TOPSCIENCE L6100) in two human PDAC cell lines (PANC-1 and AsPC-1) via CCK-8 assays to identify potential alternatives (Fig. and ).

Techniques: In Vitro, In Vivo, CCK-8 Assay, Colony Assay, Western Blot, Immunohistochemical staining, Staining

Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Agrimol B induces mitochondrial damage in PDAC cells. (A) Results of label-free quantitative proteomics after Agrimol B treatment for 24 h. (B) Western blot analysis of HADHA, TIM23, and SOD2 in PANC-1 and AsPC-1 cells. (C) Flow cytometric analysis of Fluo-4 AM accumulation in cells treated with or without 45 μmol/l Agrimol B. (D) Representative images of Fluo-4 AM accumulation in PANC-1 and AsPC-1 cells treated with or without 45 μmol/l Agrimol B for 24 h. Scale bars, 10 μm. (E) Flow cytometric analysis of mitochondrial ROS accumulation in cells treated with or without 45 μmol/l Agrimol B. (F) Representative images of mitochondrial morphology stained with Annexin V-FITC and MitoTracker Red CMXRos in PANC-1 and AsPC-1 cells treated with or without 45 μmol/l Agrimol B for 24 h. Scale bars, 10 μm. (G) Quantitative RT-PCR analysis of mtDNA copies. (H) ATP levels in PANC-1 and AsPC-1 cells treated with or without 45 μmol/l Agrimol B for 24 h. (I) Mitochondrial morphology was observed via transmission electron microscopy after treatment with or without Agrimol B for 24 h. Scale bars, 500 nm.

Article Snippet: First, we performed an unbiased, comparative analysis of therapeutic responses among 62 polyphenolic compounds (TOPSCIENCE L6100) in two human PDAC cell lines (PANC-1 and AsPC-1) via CCK-8 assays to identify potential alternatives (Fig. and ).

Techniques: Quantitative Proteomics, Western Blot, Staining, Quantitative RT-PCR, Transmission Assay, Electron Microscopy

Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Agrimol B induces PINK1/Parkin pathway-dependent mitophagy initiation in PDAC cells. (A) Western blot analysis of PINK1, Parkin, and LC3 in the mitochondria of PANC-1 and AsPC-1 cells. (B) Western blot analysis of Parkin in the mitochondria and cytoplasm of PANC-1 and AsPC-1 cells. (C) Western blot analysis of LC3 in the presence or absence of Agrimol B in the presence or absence of Mdivi-1 for 24 h. (D, E) Western blot analysis of LC3 in PDAC cells transfected with siScramble, siPINK1, or siParkin following treatment with or without Agrimol B. (F) Western blot analysis of LC3 in PANC-1 and AsPC-1 cells with or without Agrimol B in the presence or absence of wortmannin. (G-I) Immunofluorescence analysis of LC3 in PDAC cells treated with or without Agrimol B in the presence or absence of wortmannin. Scale bars, 10 μm.

Article Snippet: First, we performed an unbiased, comparative analysis of therapeutic responses among 62 polyphenolic compounds (TOPSCIENCE L6100) in two human PDAC cell lines (PANC-1 and AsPC-1) via CCK-8 assays to identify potential alternatives (Fig. and ).

Techniques: Western Blot, Transfection, Immunofluorescence

Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Agrimol B blocks autophagic flux in PDAC cells. (A, B) Western blot analysis of P62 and CTSD in PANC-1 and AsPC-1 cells treated with Agrimol B for 24 h. (C, E, F) Immunofluorescence analysis of RFP-GFP-LC3 after PANC-1 and AsPC-1 cells were transfected with RFP-GFP-LC3 for 48 h, followed by treatment with or without Agrimol B for another 24 h. Scale bars, 10 μm. (D, G-L) Immunofluorescence analysis of the colocalization of endogenous LC3 with LAMP2 after treatment with Agrimol B or rapamycin for 24 h in PANC-1 and AsPC-1 cells. Scale bars, 10 μm. (M-O) Immunofluorescence analysis of LC3 in PDAC cells treated with or without Agrimol B in the presence or absence of HCQ. Scale bars, 10 μm.

Article Snippet: First, we performed an unbiased, comparative analysis of therapeutic responses among 62 polyphenolic compounds (TOPSCIENCE L6100) in two human PDAC cell lines (PANC-1 and AsPC-1) via CCK-8 assays to identify potential alternatives (Fig. and ).

Techniques: Western Blot, Immunofluorescence, Transfection

Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Accumulation of mitophagosomes contributes to the anti-PDAC effects of Agrimol B. (A–D) CCK-8 assay of PDAC cells incubated with or without Agrimol B in the presence or absence of wortmannin and HCQ. (E–H) CCK-8 assay in PDAC cells transfected with siScramble, siPINK1, or siParkin following treatment with or without Agrimol B. (I, J) Flow cytometric analysis of PDAC cells incubated with or without Agrimol B in the presence or absence of wortmannin, HCQ.

Article Snippet: First, we performed an unbiased, comparative analysis of therapeutic responses among 62 polyphenolic compounds (TOPSCIENCE L6100) in two human PDAC cell lines (PANC-1 and AsPC-1) via CCK-8 assays to identify potential alternatives (Fig. and ).

Techniques: CCK-8 Assay, Incubation, Transfection

Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Agrimol B regulates mitophagy by downregulating MTERF3 expression. (A) Venn diagram showing the overlap of differentially expressed proteins (fold-change ≥ 1.3 or ≤ 0.76) between PANC-1 and AsPC-1 cells. (B, C) Volcano plots of DEGs identified via label-free quantitative proteomics in PANC-1 and AsPC-1 cells. (D) Western blot analysis of MTERF3 in PANC-1 and AsPC-1 cells treated with Agrimol B for 24 h. (E) Differences in MTERF3 expression between normal tissues and cancer tissues in the UCSC Xena database. (F) Kaplan-Meier analysis of MTERF3 expression and overall survival in 64 patients with PDAC. (G, H) CCK-8 assay in PDAC cells transfected with vector or oeMTERF3 following treatment with or without Agrimol B. (I) Western blot analysis of LC3 in PDAC cells transfected with vector or oeMTERF3 following treatment with or without Agrimol B. (J) Western blot analysis of PINK1 and Parkin in PDAC cells transfected with vector or oeMTERF3 following treatment with or without Agrimol B. (K) Immunohistochemical analyses of PINK1 and MTERF3 expression in PDAC tissues. Scale bars, 100 μm. (L) Correlation of the immunostaining intensities of PINK1 and MTERF3. (M) Western blot analysis of TIM23, SOD2, and HADHA in PDAC cells transfected with vector or oeMTERF3 following treatment with or without Agrimol B. (N) Molecular docking suggests that Agrimol B can bind to MTERF3 with a binding energy of -6.085 kcal/mol. (O) Western blot analysis of MTERF3 in cells treated with or without Agrimol B in the presence or absence of MG132.

Article Snippet: First, we performed an unbiased, comparative analysis of therapeutic responses among 62 polyphenolic compounds (TOPSCIENCE L6100) in two human PDAC cell lines (PANC-1 and AsPC-1) via CCK-8 assays to identify potential alternatives (Fig. and ).

Techniques: Expressing, Quantitative Proteomics, Western Blot, CCK-8 Assay, Transfection, Plasmid Preparation, Immunohistochemical staining, Immunostaining, Binding Assay

Journal: Cancer Research

Article Title: Spatial Multiomics Analyses Reveal That Diabetes Promotes Pancreatic Cancer Progression by Stimulating Cholesterol-Induced Neutrophil Extracellular Trap Formation

doi: 10.1158/0008-5472.CAN-25-2854

Figure Lengend Snippet: Inhibition of SREBP2-mediated cholesterol biosynthesis restrains CXCL1 expression and PDAC growth under DM. A, Experimental outline of murine PDAC orthotopic model. WT and db/db mice were implanted with Pan02 cells and sacrificed 3 weeks after tumor implantation. B, Representative images of tumors at the endpoint. The tumor volume and weight were compared. Scale bars, 5 mm; n = 6. C, The serum CXCL1 and cholesterol levels in the WT mice compared with the db/db mice. D, Representative immunofluorescence images and quantification of CD45 + Ly6g + and MPO + CitH3 + cells in the tumors. Scale bars, 50 µm; n = 6. Green, CD45- and CitH3-positive cells; red, Ly6g- and MPO-positive cells. E, Flow cytometric quantification of CXCR1 + and CXCR2 + neutrophils in the tumors between STZ-induced diabetic mice and control mice ( n = 5). F, Representative Western blot images and quantitative comparison of SREBP2 protein between WT and db/db mice ( n = 4). G, Representative Western blot images and quantitative comparison of SREBP2 protein between control and hyperglycemia samples ( n = 3). H, The relative mRNA expression of SREBP2 in the Panc-1 cells stimulated by high concentration of glucose versus normal glucose ( n = 3). I, The level of CXCL1 in cellular supernatant ( n = 3). J, A schematic diagram showing the experiment design. Three days after injection, mice were administered with betulin or simvastatin as indicated. K, The tumor volume and weight were compared. Scale bars, 5 mm; n = 6. L, The serum level of CXCL1 between control, betulin, and simvastatin groups. Representative immunofluorescence images and quantification of CD45 + Ly6g + and MPO + CitH3 + cells in the tumors. Scale bars, 50 µm; n = 5. M, Dotplot showing top 10 pathways enriched by upregulated genes in the DM group using KEGG analysis. N, Western blots of tumor samples showed the upregulation in phosphorylated AKT and NF-κB proteins and total AKT and NF-κB proteins. P values were determined by two-tailed ordinary one-way ANOVA with the Tukey post hoc test or Student t test. Data are represented as mean ± SEM.

Article Snippet: The mouse pancreatic cancer cell line Pan02 (RRID: CVCL_D627) was provided by were provided by Procell Life Science & Technology.

Techniques: Inhibition, Expressing, Tumor Implantation, Immunofluorescence, Control, Western Blot, Comparison, Concentration Assay, Injection, Two Tailed Test

Journal: Cancer Research

Article Title: Spatial Multiomics Analyses Reveal That Diabetes Promotes Pancreatic Cancer Progression by Stimulating Cholesterol-Induced Neutrophil Extracellular Trap Formation

doi: 10.1158/0008-5472.CAN-25-2854

Figure Lengend Snippet: DM-mediated SREBP2 increases CXCL1 secretion by activating the PI3K/AKT and NF-κB signaling pathways. A, A schematic diagram showing the experiment design. B, WT and PAD4 −/− mice were implanted with shNC Pan02 cells or SREBP2 silencing Pan02 cells and sacrificed 3 weeks after tumor implantation. Scale bars, 5 mm; n = 5. The tumor volume and weight were compared. C, The serum level of CXCL1 between groups. D, The expression of CK5, Ly6g, CXCL1, CXCR2, and CXCR1 between control and SREBP2 knockdown tumors by mIHC. Scale bars, 100 μm ( n = 5). E, Representative immunofluorescence images and quantification of MPO + CitH3 + cells in the tumors. Scale bars, 50 µm; n = 5. F, Western blots showing the upregulation in phosphorylated AKT and NF-κB proteins and total AKT and NF-κB proteins in tumor tissues ( n = 4). G, Representative Western blot images and quantitative comparison of total and phosphorylation of NF-κB and AKT proteins in shNC and shSREBP2 tumor cells ( n = 3). H, A schematic diagram showing the experiment design. I, The tumor volume, weight, and CXCL1 level were compared between shNC and shSREBP2 with or without DHMEQ treatment. J, Representative immunofluorescence images and quantification of MPO + CitH3 + cells within tumors. Scale bars, 50 µm; n = 5. K, The tumor volume, weight, and CXCL1 level were compared between shNC and shSREBP2 with or without PI3K/AKT-IN-1 treatment. L, Representative immunofluorescence images and quantification of MPO + CitH3 + cells in the tumors. Scale bars, 50 µm; n = 5. P values were determined by two-tailed ordinary one-way ANOVA with the Tukey post hoc test or Student t test. Data are represented as mean ± SEM. ns, nonsignificant.

Article Snippet: The mouse pancreatic cancer cell line Pan02 (RRID: CVCL_D627) was provided by were provided by Procell Life Science & Technology.

Techniques: Protein-Protein interactions, Tumor Implantation, Expressing, Control, Knockdown, Immunofluorescence, Western Blot, Comparison, Phospho-proteomics, Two Tailed Test

Journal: Cancer Research

Article Title: Spatial Multiomics Analyses Reveal That Diabetes Promotes Pancreatic Cancer Progression by Stimulating Cholesterol-Induced Neutrophil Extracellular Trap Formation

doi: 10.1158/0008-5472.CAN-25-2854

Figure Lengend Snippet: Cancer cell–derived cholesterol promotes NET formation through activating CXCL1/CXCR1/CXCR2 axis. A, Experimental outline of blockade of the CXCL1/CXCR1/CXCR2 axis in the orthotopic PDAC tumor model using neutralizing antibody reparixin. B, WT mice and STZ-induced diabetic mice were implanted with Pan02 cells and sacrificed 3 weeks after tumor implantation. Representative images (duplicates) of orthotopic PDAC tumors. Scale bar, 5 mm. The tumor volume and weight were compared ( n = 5). C, Representative images and quantification of Ki-67–positive cells in tumor tissues by IHC. Scale bars, 50 µm; n = 5. D, Representative immunofluorescence images and quantification of CD45 + Ly6g + and MPO + CitH3 + cells in the tumors. Scale bars, 100 µm; n = 5. Red, CD45-and MPO-positive cells; green, Ly6g- and CitH3-positive cells. E, Experimental outline of blockade of the CXCL1/CXCR1/CXCR2 axis in the orthotopic PDAC tumor model using neutralizing antibody SX-517. F, The tumor volume and weight were compared between WT, STZ, and STZ + SX517 groups. G, Representative immunofluorescence images and quantification of MPO + CitH3 + cells in the tumors. Scale bars, 50 µm. H, A schematic diagram showing the experiment design. I, The tumor volume and weight were compared between shNC and shCXCL1 groups. J, Representative immunofluorescence images and quantification of MPO + CitH3 + cells in the tumor tissues. Scale bars, 50 µm; n = 6. K, Flow cytometric quantification of CXCR1 + and CXCR2 + neutrophils in the tumors ( n = 6). L, Schematic representation of bicellular neutrophil-tumor cell coculture system. Representative immunofluorescence staining images of SYTOX to indicate NETs in the neutrophils. Scale bars, 50 µm. P values were determined by two-tailed ordinary one-way ANOVA with the Tukey post hoc test or Student t test. Data are represented as mean ± SEM.

Article Snippet: The mouse pancreatic cancer cell line Pan02 (RRID: CVCL_D627) was provided by were provided by Procell Life Science & Technology.

Techniques: Derivative Assay, Tumor Implantation, Immunofluorescence, Staining, Two Tailed Test