miapaca 2  (ATCC)

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: Copper deprivation induced by SLC31A11 knockdown triggers the upregulation of SLC7A11. ( A ) Confocal imaging of Cu-probe and Mito-tracker in the NC and SLC31A1 knockdown AsPC-1 cells. Hoechst (blue) is used as a nuclear counterstain. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. Scale bar: 2 μm. ( B ) Total Fe level in the NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( C ) Volcano plot of gene expression (the SLC31A1 knockdown [shRNA#2] versus the control; log2(fold change) ≥1; p < 0.05 between SLC31A1 knockdown and NC AsPC-1 cells. ( D ) GO analysis of differentially expressed genes between SLC31A1 knockdown (shRNA#2) and NC AsPC-1 cells. ( E-G ) Western blot analysis of the indicated protein levels in the NC and SLC31A1 knockdown AsPC-1 (E), MiaPaCa-2 (F), and CFPAC-1 (G) cells. ( H and I ) GPX4 (H) or FSP-1 activity (I) was tested in the NC and SLC31A1 knockdown AsPC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( J ) qPCR analysis of SLC31A1 and SLC7A11 gene expression in NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( K ) qPCR analysis of SLC31A1 and SLC7A11 gene expression in NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( L ) GSH level in the NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( M ) Cystine uptake level in the NC and SLC31A1 knockdown AsPC-1, MiaPaCa-2, and CFPAC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test.

Article Snippet: AsPC-1, CFPAC-1, MiaPaCa-2, SW1990, KYSE-410, NCI–H1666, SW579, and OS-RC-2 cell lines were obtained from the American Type Culture Collection (ATCC; CRL-1682, CRL-1918, CRM-CRL-1420, CRL-2172, CL-0586, CL-0845, CL-0224, CL-0177).

Techniques: Knockdown, Imaging, Gene Expression, shRNA, Control, Western Blot, Activity Assay

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: Copper deprivation induced by SLC31A1 knockdown attenuates ferroptosis. ( A ) Cell viability of AsPC-1 cells treated with different doses of RSL3, ML162, ML210, or erastin in NC and SLC31A1 knockdown AsPC-1 cells. ( B ) Cell viability of AsPC-1 cells treated with different doses of CDDP, staurosporine, paclitaxel, bortezomib, JTC-801, or elesclomol-Cu in NC and SLC31A1 knockdown AsPC-1 cells. ( C-E ) Propidium iodide (PI) staining of NC and SLC31A1 knockdown AsPC-1 (C), MiaPaCa-2 (D), and CFPAC-1 (E) cells treated with RSL3 at the indicated concentrations in the presence or absence of ferroptosis inhibitors (ferrostatin-1/Fer-1, 5 μM or desferrioxamine/DFO, 20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( F ) Lipid peroxidation of NC and SLC31A1 knockdown AsPC-1 cells treated with RSL3 (0.25 μM) for 2 h. (G) Transmission electron microscopy of NC and SLC31A1 knockdown AsPC-1 cells treated with RSL3 (2.5 μM). Scale bar: 2 μm or 500 nm. ( H ) Confocal imaging of Cu-probe and Mito-tracker in the NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1. Hoechst (blue) is used as a nuclear counterstain. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. Scale bar: 2 μm. ( I ) Western blot analysis of lysates from NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1. ( J ) Cell viability of NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1, followed by treatment with different doses of RSL3 or erastin. ( K ) PI staining of NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or a vector encoding SLC31A1, followed by treatment with RSL3 at the indicated concentrations in the presence or absence of Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( L ) Western blot analysis of lysates from SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or SLC31A1 WT or SLC31A1 M154A plasmid. ( M ) PI staining of SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with a blank vector or SLC31A1 WT or SLC31A1 M154A plasmid, followed by treatment with RSL3 at the indicated concentrations in the presence or absence of Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test.

Article Snippet: AsPC-1, CFPAC-1, MiaPaCa-2, SW1990, KYSE-410, NCI–H1666, SW579, and OS-RC-2 cell lines were obtained from the American Type Culture Collection (ATCC; CRL-1682, CRL-1918, CRM-CRL-1420, CRL-2172, CL-0586, CL-0845, CL-0224, CL-0177).

Techniques: Knockdown, Staining, Transmission Assay, Electron Microscopy, Imaging, shRNA, Transfection, Plasmid Preparation, Western Blot

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: Pharmacological induction of copper deprivation upregulates SLC7A11 and suppresses ferroptosis. ( A ) Western blot analysis of lysates from AsPC-1 cells treated with the indicated concentrations of TM for 24 h. ( B ) SLC7A11 mRNA level in AsPC-1 cells treated with TM at the indicated concentrations for 24 h. ( C ) Western blot analysis of lysates from AsPC-1 cells treated with TEPA at the indicated concentrations for 24 h. ( D ) SLC7A11 mRNA level in AsPC-1 cells treated with TEPA at the indicated concentrations for 24 h. ( E ) GSH level in the AsPC-1 cells treated with TM (50 μM) or TEPA (1 mM) for 24 h. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( F ) Cystine uptake level in the AsPC-1 cells treated with TM (50 μM) or TEPA (1 mM) for 24 h. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( G ) Cell viability of PDAC cells treated with RSL3 in the presence or absence of TM (50 μM) or TEPA (1 mM) for 24 h. ( H-J ) Propidium iodide (PI) staining of AsPC-1 (H), MiaPaCa-2 (I), and CFPAC-1 (J) cells treated with RSL3 at the indicated concentrations in the presence or absence of TM (50 μM) or TEPA (1 mM) with or without Fer-1 (5 μM) or DFO (20 μM) for 6 h. Quantification of PI-positive cells was shown. Mean ± SD, n = 3. Statistical significance was determined using a two-way ANOVA test. ( K ) Lipid peroxidation of AsPC-1 cells treated with RSL3 (0.25 μM) in the presence or absence of TM (50 μM) or TEPA (1 mM) for 2 h. ( L ) Cell viability of KYSE-410, NCI–H1666, SW579, or OS-RC-2 cells treated with RSL3 at the indicated concentrations in the presence or absence of TM (50 μM) for 12 h.

Article Snippet: AsPC-1, CFPAC-1, MiaPaCa-2, SW1990, KYSE-410, NCI–H1666, SW579, and OS-RC-2 cell lines were obtained from the American Type Culture Collection (ATCC; CRL-1682, CRL-1918, CRM-CRL-1420, CRL-2172, CL-0586, CL-0845, CL-0224, CL-0177).

Techniques: Western Blot, Staining

Journal: Redox Biology

Article Title: Copper deprivation reprograms antioxidant defense to suppress ferroptosis via SLC7A11

doi: 10.1016/j.redox.2026.104130

Figure Lengend Snippet: Copper deprivation upregulates SLC7A11 and activates AMPK-NRF2 axis. ( A ) Volcano plot of NRF2 target genes (shSLC31A1 VS shNC) generated using the data from C. ( B , C ) ChIP analyses of NRF2 binding in the SLC7A11 promoter in NRF2 knockdown AsPC-1 cells (B) or SLC31A1 knockdown AsPC-1 cells (C). ( D-G ) The oxygen consumption rate (OCR) in NC and SLC31A1 knockdown AsPC-1 cells by Seahorse assay (n = 8/group). Quantification of basal respiration (E) and ATP-linked respiration (F), the uncoupler FCCP (maximal) or the electron transport inhibitor antimycin A-baseline (G) in NC and SLC31A1 knockdown AsPC-1 cells. Mean ± SD, n = 8. Statistical significance was determined using a one-way ANOVA test. ( H ) COX activity level in the NC and SLC31A1 knockdown AsPC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( I–K ) ATP level in the NC and SLC31A1 knockdown AsPC-1(I), MiaPaCa-2 (J), and CFPAC-1 (K) cells. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( L ) ATP level in the AsPC-1 cells treated with TM (50 μM) or TEPA (1 mM) for 24 h. Mean ± SD, n = 3. Statistical significance was determined using a one-way ANOVA test. ( M ) AMP/ATP ratio detected by HPLC in NC and SLC31A1 knockdown AsPC-1 cells. Mean ± SD, n = 3. Statistical significance was determined using an unpaired T test. ( N ) Western blot analysis of lysates from NC and SLC31A1 knockdown AsPC-1 cells. ( O ) Western blot analysis of lysates from AsPC-1 cells treated with TM at the indicated concentrations for 24 h. ( P ) Western blot analysis of lysates from NC and SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with blank vector or vector encoding SLC31A1. ( Q ) Western blot analysis of lysates from SLC31A1 knockdown (shRNA#2) AsPC-1 cells transfected with blank vector or of SLC31A1 WT- or SLC31A1 M154A.

Article Snippet: AsPC-1, CFPAC-1, MiaPaCa-2, SW1990, KYSE-410, NCI–H1666, SW579, and OS-RC-2 cell lines were obtained from the American Type Culture Collection (ATCC; CRL-1682, CRL-1918, CRM-CRL-1420, CRL-2172, CL-0586, CL-0845, CL-0224, CL-0177).

Techniques: Generated, Binding Assay, Knockdown, Activity Assay, Western Blot, shRNA, Transfection, Plasmid Preparation

Journal: bioRxiv

Article Title: DHHC7 palmitoylates KRAS4A and promotes mutant KRAS-driven pancreatic cancers

doi: 10.64898/2026.03.31.715686

Figure Lengend Snippet: ( a, b ) DHHC7 promotes pERK and 2-D cell growth of pancreatic cancer cells. ZDHHC7 was knockdown using siRNA in four mutant KRAS-driven pancreatic cancer cell lines (AsPC1, MiaPaca-2, PANC0327 and Tu8988s) ( a ) and one non-KRAS driven pancreatic cancer cell line (BxPC3) ( b ). pERK was measured using western blot and 2-D cell growth was measured using Cell Titer Glow after 48-72 hrs. ( c-d ) ZDHHC7 knockout affects pERK, 2-D cell growth, KRAS4A S-palmitoylation in MiaPaca-2 cell line. ZDHHC7 was CRISPR knockout and pERK levels ( c ) were detected using western blot and 2D cell growth was measured after 72 hrs. The endogenous S-acylation level of KRAS4A was detected using ABE ( d ). ( e ) ZDHHC7 was CRISPR knockout in AsPC1 cell line and pERK levels were detected using western blot. ( f-h ) ZDHHC7 knockout suppresses MiaPaca-2 xenograft tumor growth. Pancreatic cancer xerograph was performed using MiaPaca-2 cells with control or ZDHHC7 CRISPR knockout. The tumor volume was measured and quantified (f). The mice were dissected, and the final tumors were shown ( g ) and the tumor weights were quantified ( h ). ( i-k ) ZDHHC7 knockout suppresses AsPC1 xenograft tumor growth. ZDHHC7 CRISPR knockout was performed in AsPC1 cell line and similar xenograft experiment was performed. Tumor volume (i), tumors (j), tumor weight (k) are shown. Quantification data are shown as mean ± SD. ns, not significant. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Tumors were established by subcutaneously injecting MiaPaca-2 cells (3 × 10^6 cells per animal), PANC 0327 cells (1 x 10^6 cells per animal), AsPC1 cells (1 x 10^6 cells per animal) and BxPC3 cells (1 x 10^6 cells per animal) into 4- to 6-week-old male NSG mice (Jackson Laboratory, Bar Harbor, US).

Techniques: Knockdown, Mutagenesis, Western Blot, Knock-Out, CRISPR, Control

Journal: Materials Today Bio

Article Title: Escape from cell uptake: Drug-Free cancer therapeutics regulated by hydrophobicity and negative charge

doi: 10.1016/j.mtbio.2025.102752

Figure Lengend Snippet: Cytotoxicity and Cell adsorption property of PVA-U varying G.D. (a) IC 50 values of MIAPaCa-2 cells treated with UDCA, PVA-U3, PVA-U15, and PVA-U25 for 24 h incubation at pH 7.4 and pH 6.5 (n = 3). (b) Cell viability of MIAPaCa-2 cells treated with 10 μg mL −1 PVA-U15 at pH 7.4 and pH 6.5 for 0, 4, 12, and 24 h incubation (n = 3). (c) Confocal images of MIAPaCa-2 cells treated with Cell tracker deep red and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 7.4 and pH 6.5 for 12 h incubation. Cell tracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 100 μm and 50 μm, respectively. Quantification of intracellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 24 h at pH 7.4 (d) and pH 6.5 (e) (n = 11). Quantification of pericellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 4 h at pH 7.4 (f) and pH 6.5 (g) (n = 11). (h) Confocal images of MIAPaCa-2 cells treated with lysotracker and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 6.5 for 4 h incubation. Lysotracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 50 μm and 10 μm, respectively. Statistical analysis was performed using unpaired two-tailed Student's t -test. Data are presented as mean ± S.D. ( N.S. : no significant difference, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: MIAPaCa-2 cells, HT-29 cells, and A549 cells were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Adsorption, Incubation, Fluorescence, Two Tailed Test

Journal: Materials Today Bio

Article Title: Escape from cell uptake: Drug-Free cancer therapeutics regulated by hydrophobicity and negative charge

doi: 10.1016/j.mtbio.2025.102752

Figure Lengend Snippet: Antitumor efficacy of PVA-U15. (a) Treatment schedule for MIAPaCa-2 tumors. BALB/c nude mice were inoculated with MIAPaCa-2 cells on day 0. The tumor-bearing mice were randomized and treated with UDCA (144 μM), PVA (10 μg mL −1 ) or PVA-U15 (10 μg mL −1 ). 100 μL of each solution was intratumorally injected 5 days per week from day 9 to day 21. (b) Tumor growth in mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). (c) Body weight of mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). Statistical analysis was performed using Tukey test. Data are presented as mean ± S.D. (∗ p < 0.05).

Article Snippet: MIAPaCa-2 cells, HT-29 cells, and A549 cells were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Injection