pdac human cell lines aspc 1 p53 null  (ATCC)

Journal: Cell Death & Disease

Article Title: Chemotherapy enhances HMGA1 secretion through the mutant p53-CK2 axis in pancreatic ductal adenocarcinoma cells

doi: 10.1038/s41419-025-08082-1

Figure Lengend Snippet: A Differentially secreted proteins detected by MS analysis in the CM of PANC-1 cells transiently transfected with si TP53 R273H or siScramble. The 25 proteins significantly downsecreted are indicated by the light lavender-shaded rectangle underlaid on the plot. Vertical dashed lines indicate log2 fold change = ±0.5. Horizontal dashed line indicates the cut-off p-value p = 0.05. TPM1, CSTN1, HMGA1 and CP2R1 proteins are marked on the plot. B Differentially secreted proteins detected by MS analysis in the CM of AsPC-1 cells transiently transfected with TP53 R273H or MOCK plasmid. The 83 proteins significantly hypersecreted are indicated by the light teal-shaded rectangle underlaid on the plot. Among them, TPM1, CSTN1, HMGA1 and CP2R1 proteins are specifically highlighted. Vertical dashed lines indicate log2 fold change = ±0.5. Horizontal dashed line indicates the cut-off p-value p = 0.05. C Venn diagram indicating the overlaps of the differentially mut or wt p53-dependent secreted proteins detected by MS analysis. D Histogram showing 4 proteins (Tropomyosin 1 (TPM1), Calsyntenin 1 (CSTN1), High Mobility Group A1 (HMGA1), Cytochrome P450 Family 2 Subfamily R Member 1 (CP2R1)) hypersecreted by AsPC-1 (p53-null) cells overexpressing TP53 R273H and downsecreted by PANC-1 cells after KD of the same hot-spot mutp53 isoform. E UMAP visualization of all identified cell types present in the pancreatic microenvironment subset by disease state: Adjacent Normal ( n = 3), Healthy ( n = 6) and Tumor ( n = 16). Data source: Pancreatic Tissue Single Cell Atlas. F UMAP visualizations showing HMGA1 expression across the major cell populations subset by disease state (Adjacent Normal, Healthy, Tumor). Data source: Pancreatic Tissue Single Cell Atlas. G HMGA1 gene expression level in tumor derived epithelial cells compared to adjacent normal or healthy epithelial cells (number of cells: 892 Adj.Normal; 14,380 Healthy; 9484 Tumor). Data source: Pancreatic Tissue Single Cell Atlas. H HMGA1 gene expression level in tumor compared to normal tissue. Data source: GEPIA database. * p < 0.01. I HMGA1 expression correlates with the mutational status of TP53 ( n = 66 no-mutation, n = 62 missense mutations). Data source: cBioportal database, TCGA PanCancer Atlas . (Wilcoxon test). **** p < 0.0001. J HMGA1 expression level in PDAC patient with TP53 mut ( n = 24 no-mutation, n = 43 missense mutations Data source: cBioportal database, QCMG Nature 2016 . (Wilcoxon test). **** p < 0.0001. K Kaplan-Meier (KM) plot of survival probability (log-rank test) for PDAC patients only as obtained from KM-plotter database using the default parameters. L Kaplan-Meier survival plot after PDAC patients’ stratification for tumor stage (S2, S3, S4) in KM plotter database showing HMGA1 expression is a prognostic factor in advanced pancreatic cancer. M Volcano plot of differential gene expression (DGE) analysis for metastatic vs primary tumors using the microarray dataset GSE71729 showing HMGA1 is significantly highly expressed in metastatic patients (Log2FC = 2.383837; −log10(Adj. p-value) = 10.47756). Red highlights indicate significant regulated genes; black highlights indicate non-significant genes. Vertical dashed lines indicate log2 fold change = ±1. Horizontal dashed line indicates p = 0.01. N Immunoblot validation of HMGA1 KO in human PANC-1 cell line. KO denotes HMGA1 KO cells, while the minus sign (-) represents the parental cells. Vinculin was used as a loading control. O Representative images, with corresponding magnifications, of PANC-1 (top) and HMGA1-KO PANC-1 (bottom) cells invading through Matrigel-coated transwell inserts (8 μm pore size) after 24 h of incubation at 37 °C. Right panel: bar plot quantification of the percentage of invasive cells. Data are presented as mean ± SD (n = 4). (Unpaired t-test). **p < 0.01. P Tumor volume (mm 3 ) from subcutaneous injection of either PANC-1 or HMGA1 KO PANC-1 cells in immunodeficient mice. Data plotted are mean tumor volumes + SEM ( n = 6 for each cohort). (Two-way ANOVA). ****p < 0.0001. Q Individual tumor volumes + SEM at the endpoint (Unpaired t-test). *p < 0.05.

Article Snippet: The PDAC human cell lines AsPC-1 (p53-null) and PANC-1 ( TP53 R273H ) were purchased from the American Type Culture Collection (ATCC).

Techniques: Transfection, Plasmid Preparation, Expressing, Gene Expression, Derivative Assay, Mutagenesis, Microarray, Western Blot, Biomarker Discovery, Control, Pore Size, Incubation, Injection

Journal: Cell Death & Disease

Article Title: Chemotherapy enhances HMGA1 secretion through the mutant p53-CK2 axis in pancreatic ductal adenocarcinoma cells

doi: 10.1038/s41419-025-08082-1

Figure Lengend Snippet: A Immunoblot of secreted HMGA1 protein validating the MS-data. Amido black staining (a.b.) was used as loading control for WB. B Immunoblot of HMGA1 protein in different human PDAC cell lines. C Immunoblot of secreted HMGA1 protein in different human PDAC cell lines. D Immunoblot validation of TP53 KO in human PANC-1 cell line. KO denotes TP53 KO cells, while the minus sign (−) represents the parental cells. Vinculin was used as a loading control. E Immunoblot of secreted HMGA1 protein in PaCa3, PANC-1 and TP53 KO PANC-1 cells. F Cell growth percentage measured by cristal violet assay in HMGA1 KO PANC-1 cells cultured for 48 h with PANC-1 CM or HMGA1 KO PANC-1 CM. (Unpaired t-test). *** p < 0.001. G Cell growth percentage measured by cristal violet assay in p53-null AsPC-1 cells cultured for 48 h with R273H CM or MOCK CM after the addition of anti-HMGA1 antibody (0.226 µg/µl, 1:100 in growth medium) or the IgG Isotype Control (Cell signaling, 5742). (Two-way ANOVA). *p < 0.05, **p < 0.01. H Immunoblot analysis of HMGA1 protein in PANC-1 CM alongside different µg of rHMGA1 protein. On the right, the slope obtained from linear regression analysis of the average adjusted total band intensity values + SD (arbitrary units, a.u.) of immunoreactivity corresponding to rHMGA1. The adjusted total band intensity values were derived from densitometric analysis of the immunoreactive bands for HMGA1 secreted by PANC-1 cells, as well as rHMGA1 used as a standard with increasing sample loads. Data were analyzed using Image Lab Software (Bio-Rad, version 6.1.0 build 7). The blue square on the slope represents the value corresponding to the secreted HMGA1. I Cell growth percentage measured by cristal violet assay in HMGA1 KO PANC-1 cells cultured for 72 h after treatment with different doses of rHMGA1 protein. (One-way ANOVA). ****p < 0.0001.

Article Snippet: The PDAC human cell lines AsPC-1 (p53-null) and PANC-1 ( TP53 R273H ) were purchased from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Staining, Control, Biomarker Discovery, Cell Culture, Derivative Assay, Software

Journal: Cell Death & Disease

Article Title: Chemotherapy enhances HMGA1 secretion through the mutant p53-CK2 axis in pancreatic ductal adenocarcinoma cells

doi: 10.1038/s41419-025-08082-1

Figure Lengend Snippet: A Immunoblot of p-p53, p53 and HMGA1 proteins in PANC-1 cells after treatment with different anti-cancer drugs at sublethal doses (1 µM gemcitabine (GEM), 5 µM 5-fluorouracil (5-FU), 1 µM oxaliplatin (OXA) and 5 µM irinotecan (IRI)). B Immunoblot of HMGA1 protein in PANC-1 cells secretome after treatment with different anti-cancer drugs at sublethal doses. C Bar charts depict A.I. (a.u.) of HMGA1 secreted by PANC-1 cells with or without 1 µM GEM treatment versus a.b. analyzed using Image Lab Software (Bio-Rad, version 6.1.0 build 7). Data plotted are mean of seven independent experiments ± SD. (Unpaired t-test). *p < 0.05. D qPCR showing TP53 expression upon sublethal dose GEM treatment of PANC-1 cells for 6, 10, 24 and 48 h. ***p < 0.001, ****p < 0.0001. E qPCR showing HMGA1 expression upon sublethal dose of GEM treatment of PANC-1 cells for 6, 10, 24, and 48 h. F Immunoblot of p-p53, p53, and HMGA1 proteins in PANC-1 cells upon sublethal dose GEM treatment for 6, 10, 24 and 48 h. G Bar charts of PaCa3, Hs776t, PANC-1 and SUIT-2 cells viability (PI-/Ann V-) after 24 h treatment with 1 µM GEM. H Immunoblot of p53 and HMGA1 proteins in two cell lines carrying TP53 w t (PaCa3 and Hs776t) and two cell lines harboring TP53 R273H (PANC-1 and SUIT-2) after being treated or not with 1 µM of GEM. I Immunoblot of HMGA1 in human PDAC cells secretome after 1 µM GEM treatment.

Article Snippet: The PDAC human cell lines AsPC-1 (p53-null) and PANC-1 ( TP53 R273H ) were purchased from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Software, Expressing