Journal: Cancer Research Communications

Article Title: Acquisition of Resistance to RAS Inhibition Is Associated with the Upregulation of Macropinocytosis through Both PI3K-Dependent and -Independent Signaling

doi: 10.1158/2767-9764.CRC-25-0731

Figure Lengend Snippet: Upregulated macropinocytosis in RAS inhibitor-resistant cell lines is associated with increased uptake of and sensitivity to albumin-bound paclitaxel compared with free drug. A, Schematic diagram highlighting the differential routes through which free paclitaxel and albumin-bound nab-paclitaxel enter the cell. B, Representative images of macropinosomes labeled with FITC-BSA (green) and nuclear DAPI stain (blue) in indicated parental or MRTX1133- (G12Di-R) or RMC7797-resistant (RASi-R) KRAS -mutant PDAC cells. Images are representative of 10 fields of view analyzed in each of two independent experiments. Scale bars, 20 μm. C, Quantification of median intensity of TMR-BSA fluorescence in indicated MRTX1133- (G12Di-R) or RMC7797-resistant (RASi-R) KRAS -mutant PDAC cell lines plotted relative to matched parental cell line, with each individual data point representing one field containing at least 10 analyzed cells. Data are presented as the mean ± SEM of three independent experiments. ***, ρ < 0.001, and ****, ρ < 0.0001, by the unpaired Student t test, comparing against the parental cell line. D, Representative images of clonogenic growth assays following treatment of matched parental and either MRTX1133- (G12Di-R) or RMC7977-resistant (RASi-R) PANC-1 or HPAC cell lines treated with nab-paclitaxel for 12–16 days. E, Relative colony number of matched parental and either MRTX1133- (G12Di-R) or RMC7977-resistant (RASi-R) PDAC cell lines treated with nab-paclitaxel (left) or paclitaxel (right) for 12–16 days. Data are presented as the mean ± SEM of three independent experiments. F, GI 50 values from data plotted in ( E ) for nab-paclitaxel treated lines (top) and paclitaxel treated lines (bottom). *, ρ < 0.05; **, ρ < 0.01; ***, ρ < 0.001, by the unpaired Student t test, comparing against the parental cell line. [ A, Created in BioRender. Robb, R. (2026) https://BioRender.com/nyleshf .]

Article Snippet: RMC7977 (HY-156498), trametinib (HY-10999), paclitaxel (HY-B0015), nab-paclitaxel (HY-P99974), pictilisib (HY-50094), defactinib (HY-12289), and IAG933 (HY-153811) were purchased from MedChemExpress.

Techniques: Labeling, Staining, Mutagenesis, Fluorescence

Journal: Cell reports

Article Title: Cisplatin resistance in an ovarian cancer model is mediated by microtubule dynamics regulator TPPP3 in synergy with tubulin code rewiring

doi: 10.1016/j.celrep.2026.117414

Figure Lengend Snippet: (A) Principal-component analysis based on the RNA-seq data from cisplatin-sensitive, -resistant, and -re-sensitized ovarian cancer cells reveal distinct transcriptional signatures among the three cell lines. (B) Heatmap representing differentially expressed genes in the three cell lines (FDR < 0.01, absolute log FC > 2) identifies TPPP3 as one of the top genes up-regulated in resistant cells compared to sensitive or re-sensitized cells. (C) Venn diagrams depicting the number of genes differentially expressed. (D) Log2 fold-change difference between the mRNA levels of the resistant versus sensitive group and re-sensitized versus resistant group. (E) Bright-field and immunofluorescence images of sensitive, resistant, and re-sensitized ovarian cancer cells stained with DAPI and Glu-tubulin. Scale bars, 20 μm. (F) Immunofluorescence images of sensitive, resistant, and re-sensitized ovarian cancer cells stained with DAPI and TPPP3 show cytoplasmic localization of TPPP3. Scale bars, 20 μm.

Article Snippet: Similarly, paclitaxel-stabilized cisplatin resensitized (CP-RS) microtubules containing 97% CP-RS tubulin, 1% biotin brain tubulin, and 2% 647-Hilyte tubulin and paclitaxel-stabilized cisplatin resistant (CP-R) microtubules containing 99% CP-R tubulin, 1% biotin brain tubulin (Cytoskeleton, cat no. T333P) were immobilized on a glass surface using neutravidin.

Techniques: Analysis, RNA Sequencing, Immunofluorescence, Staining

Journal: Cell reports

Article Title: Cisplatin resistance in an ovarian cancer model is mediated by microtubule dynamics regulator TPPP3 in synergy with tubulin code rewiring

doi: 10.1016/j.celrep.2026.117414

Figure Lengend Snippet: (A) Schematic of interference reflection microscopy (IRM)-based microtubule dynamics assay. (B) Kymographs showing microtubule groth at 10 μM brain tubulin in the presence of 5 and 10 μM cisplatin; GMPCPP-stabilized seeds highlighted in blue. Scale bars, 5 μm (horizontal) and 5 min (vertical). (C–E) Microtubule plus-end growth rates (C), microtubule plus-end catastrophe rates (D), and microtubule lengths (E) at 10 μM porcine brain tubulin with different cisplatin concentrations. Error bars, SD. (F) Catastrophe rates as a function of microtubule growth rates in the absence (gray) or presence (red) of cisplatin; cisplatin concentration indicated above data points. Error bars, SEM; for (C) through (F) n values indicated in parentheses throughout from 8, 4, 5, 7, and 6 independent experiments on different days for 0, 1, 2.5, 5, and 10 μM cisplatin, respectively; **** p < 0.0001, ** p < 0.01, * p < 0.05, and ns, non-significant, as determined by one-way ANOVA using Tukey’s multiple comparison test. (G) TIRF microscopy images of EB1-GFP (300 nM, green) on brain microtubules in the absence or presence of 5 μM cisplatin. Scale bars, 1 μm. (H) Plus-end growth rates of microtubules with and without cisplatin in the presence of EB1-GFP. The growth rates of microtubules within 20% of the control mean are highlighted in gray and were used in the comet profile analysis. (I) Averaged fluorescent intensity profiles of EB1-GFP comets on microtubules with and without cisplatin. (J) Comet profile parameters. Comet profiles were fitted using a Gaussian (−1.5 to 0 μm) and a single exponential function (0–1.5 μm) .

Article Snippet: Similarly, paclitaxel-stabilized cisplatin resensitized (CP-RS) microtubules containing 97% CP-RS tubulin, 1% biotin brain tubulin, and 2% 647-Hilyte tubulin and paclitaxel-stabilized cisplatin resistant (CP-R) microtubules containing 99% CP-R tubulin, 1% biotin brain tubulin (Cytoskeleton, cat no. T333P) were immobilized on a glass surface using neutravidin.

Techniques: Microscopy, Concentration Assay, Comparison, Control, Analysis

Journal: Cell reports

Article Title: Cisplatin resistance in an ovarian cancer model is mediated by microtubule dynamics regulator TPPP3 in synergy with tubulin code rewiring

doi: 10.1016/j.celrep.2026.117414

Figure Lengend Snippet: (A) Kymographs showing microtubule growth at 6 μM porcine brain tubulin in the presence of paclitaxel alone, cisplatin alone, and in combination with paclitaxel at different concentrations; GMPCPP-stabilized seed highlighted in blue. Scale bars, 5 μm (horizontal) and 5 min (vertical). (B and C) Microtubule plus-end catastrophe rates (B) and lengths (C) at 6 μM of porcine brain tubulin in the presence of paclitaxel alone, cisplatin alone, and in combination with paclitaxel at different concentrations. Error bars, SD; n values shown in parentheses throughout, growth events for (B) and microtubules for (C) from 2, 3, 2, 2, 2, 2, and 2 independent experiments on different days for control, 5 μM cisplatin, 10 nM paclitaxel, 10 nM paclitaxel and 5 μM cisplatin, 20 nM paclitaxel, 20 nM paclitaxel and 5 μM cisplatin, 100 nM paclitaxel, and 100 nM paclitaxel and 5 μM cisplatin, respectively. **** p < 0.0001, ** p < 0.01, * p < 0.1, and ns, non-significant, as determined by one-way ANOVA using Tukey’s multiple comparison test. (D) Cytotoxicity assay of sensitive cells treated with increasing concentration of cisplatin in the presence of 0, 5, or 50 nM paclitaxel, n = 2 independent experiments on different days. (E) IC50 values for the data shown in (D). * p < 0.05 and ** p < 0.005, as determined by one-way ANOVA using Tukey’s multiple comparison test.

Article Snippet: Similarly, paclitaxel-stabilized cisplatin resensitized (CP-RS) microtubules containing 97% CP-RS tubulin, 1% biotin brain tubulin, and 2% 647-Hilyte tubulin and paclitaxel-stabilized cisplatin resistant (CP-R) microtubules containing 99% CP-R tubulin, 1% biotin brain tubulin (Cytoskeleton, cat no. T333P) were immobilized on a glass surface using neutravidin.

Techniques: Control, Comparison, Cytotoxicity Assay, Concentration Assay

Journal: Cell reports

Article Title: Cisplatin resistance in an ovarian cancer model is mediated by microtubule dynamics regulator TPPP3 in synergy with tubulin code rewiring

doi: 10.1016/j.celrep.2026.117414

Figure Lengend Snippet: (A) Kymographs showing microtubule growth at 6 μM tubulin purified from cisplatin-sensitive, -resistant, and -re-sensitized cells; GMPCPP-stabilized seeds highlighted in blue. Scale bars, 5 μm (horizontal) and 5 min (vertical). (B–D) Microtubule plus-end growth rates (B), microtubule plus-end catastrophe rates (C), and microtubule lengths (D) at 6 μM of tubulin purified from sensitive (magenta), resistant (cyan), and re-sensitized (orange) cells. Error bars, SD. n values indicated in parentheses throughout from 2 independent experiments on different days for each condition. Tubulin purified from an independent second preparation shows similar dynamic parameters ( – ). Data points collected from independent experiments shown in different shades. **** p < 0.0001, *** p = 0.01, * p < 0.05, and ns, non-significant, as determined by one-way ANOVA using Tukey’s multiple comparison test. (E and F) Scatterplots of microtubule growth rate (E) and microtubule growth distance (F) obtained by tracking EB3-mScarlet in cisplatin-sensitive, -resistant, and -re-sensitized cells. Number of EB3 comets and cells analyzed indicated on the graphs. n 3 independent experiments on different days. Data points collected on different days and from independent experiments are shown in different shades. Error bars, SD. **** p < 0.0001 and ns, non-significant, as determined by one-way ANOVA using Tukey’s multiple comparison test.

Article Snippet: Similarly, paclitaxel-stabilized cisplatin resensitized (CP-RS) microtubules containing 97% CP-RS tubulin, 1% biotin brain tubulin, and 2% 647-Hilyte tubulin and paclitaxel-stabilized cisplatin resistant (CP-R) microtubules containing 99% CP-R tubulin, 1% biotin brain tubulin (Cytoskeleton, cat no. T333P) were immobilized on a glass surface using neutravidin.

Techniques: Purification, Comparison

Journal: Cell reports

Article Title: Cisplatin resistance in an ovarian cancer model is mediated by microtubule dynamics regulator TPPP3 in synergy with tubulin code rewiring

doi: 10.1016/j.celrep.2026.117414

Figure Lengend Snippet: (A–C) Kymographs at 6 μM tubulin purified from cisplatin (CP)-sensitive (A), -resistant (B), and -re-sensitized (C) cells in the presence of 1 μM TPPP3, 5 μM CP, and 1 μM TPPP3 with 5 μM CP. GMPCPP-stabilized seeds highlighted in blue. Scale bars, 5 μm (horizontal) and 5 min (vertical). (D–F) Microtubule plus-end growth rates (D), microtubule plus-end catastrophe rates (E), and microtubule lengths (F) at 6 μM tubulin purified from CP-sensitive (magenta), -resistant (cyan), and -re-sensitized (orange) cells in the presence of 1 μM TPPP3, μM CP, and 1 μM TPPP3 with 5 μM CP. Error bars, SD; n values indicated in parentheses throughout from 2 independent experiments on different days for each condition. Data points collected from independent experiments shown in different shades. **** p < 0.0001, *** p < 0.001, ** p < 0.01, and ns, non-significant, as determined by one-way ANOVA using Tukey’s multiple comparison test.

Article Snippet: Similarly, paclitaxel-stabilized cisplatin resensitized (CP-RS) microtubules containing 97% CP-RS tubulin, 1% biotin brain tubulin, and 2% 647-Hilyte tubulin and paclitaxel-stabilized cisplatin resistant (CP-R) microtubules containing 99% CP-R tubulin, 1% biotin brain tubulin (Cytoskeleton, cat no. T333P) were immobilized on a glass surface using neutravidin.

Techniques: Purification, Comparison

Journal: Cell reports

Article Title: Cisplatin resistance in an ovarian cancer model is mediated by microtubule dynamics regulator TPPP3 in synergy with tubulin code rewiring

doi: 10.1016/j.celrep.2026.117414

Figure Lengend Snippet: (A) Soft-agar cytotoxicity assay of non-targeting CRISPR-resistant cells (NTCs) and TPPP3-KO cells treated with 25.6 μM cisplatin shows re-sensitization of cisplatin-resistant 1A9CP80 cells to cisplatin when TPPP3 is knocked out; n = 3 independent experiments for each KO line, ** p < 0.005, and ns, non-significant, as determined by one-way ANOVA using Tukey’s multiple comparison test. (B–D) Microtubule growth rates in response to cisplatin addition for cisplatin-resistant cells (B), TPPP3-KO cells (C), and cisplatin-sensitive cells (D). PBS buffer control shown as a discontinuous line. For cisplatin treatment, n = 31, 30, 29, and 24 resistant cells at 0, 1, 2, and 4 h, respectively (75,609, 68,043, 56,605, and 30,388 EB3 comets analyzed); n = 31, 31, 30, and 17 TPPP3-KO1 cells at 0, 1, 2, and 4 h, respectively (66,725, 43,038, 34,115, and 18,002 EB3 comets analyzed); n = 31, 31, 30, and 17 cells for TPPP3-KO2 cells at 0, 1, 2, and 4 h, respectively (54,462, 46,797, 42,793, and 32,512 EB3 comets analyzed); n = 25, 25, 21, and 17 sensitive cells at 0, 1, 2, and 4 h, respectively (29,755, 28,145, 22,020, and 13632 EB3 comets analyzed). For the PBS control, n = 30, 30, 30, and 30 resistant cells at 0, 1, 2, and 4 h, respectively (70,276, 70877, 64,407, and 50,741 EB3 comets analyzed); n = 37, 37, 35, and 33 TPPP3-KO1 cells at 0, 1, 2, and 4 h, respectively (62,200, 57,133, 40,895, and 34,201 EB3 comets analyzed); n = 36, 36, 32, and 32 TPPP3-KO2 cells at 0, 1, 2, and 4 h, respectively (74,188, 73,525, 63,872, and 53,925 EB3 comets); n = 27, 27, 27, and 25 sensitive cells at 0, 1, 2, and 4 h, respectively (25,122, 24,807, 23,841, and 19,983 EB3 comets analyzed); n.s., nonsignificant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, as determined by Mann-Whitney test. (E) Soft-agar cytotoxicity assay of 1A9-activation (1A9-ACT) and 1A9-TPPP3 activated (1A9-TPPP3) cells treated with 2.6 μM cisplatin reveals increased resistance of cisplatin-sensitive cells to cisplatin when TPPP3 is overexpressed. No cisplatin-sensitive cells survive at concentrations higher than 2.6 μM cisplatin. n = 3 independent experiments on different days. *** p < 0.0005, **** p < 0.0001, and ns, non-significant, as determined by one-way ANOVA using Tukey’s multiple comparison test. (F) Soft-agar cytotoxicity assay of non-targeting CRISPR-resistant cells (NTCs) and TPPP3-KO cells treated with 2.56 μM cisplatin shows re-sensitization of cisplatin-resistant OVCAR8-CP5 cells to cisplatin when TPPP3 is knockedout. n = 3 independent experiments. **** p < 0.0001, *** p < 0.0005, ns, not significant, as determined by one-way ANOVA using Tukey’s multiple comparison test. (G) Overall survival probability curves of 1,409 ovarian cancer patients treated with chemotherapy show poor overall survival probability in patients with high TPPP3 expression. Red and black lines indicate patients with high and low TPPP3 expression, respectivly.

Article Snippet: Similarly, paclitaxel-stabilized cisplatin resensitized (CP-RS) microtubules containing 97% CP-RS tubulin, 1% biotin brain tubulin, and 2% 647-Hilyte tubulin and paclitaxel-stabilized cisplatin resistant (CP-R) microtubules containing 99% CP-R tubulin, 1% biotin brain tubulin (Cytoskeleton, cat no. T333P) were immobilized on a glass surface using neutravidin.

Techniques: Cytotoxicity Assay, CRISPR, Comparison, Control, MANN-WHITNEY, Activation Assay, Expressing

Journal: Cell reports

Article Title: Cisplatin resistance in an ovarian cancer model is mediated by microtubule dynamics regulator TPPP3 in synergy with tubulin code rewiring

doi: 10.1016/j.celrep.2026.117414

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Similarly, paclitaxel-stabilized cisplatin resensitized (CP-RS) microtubules containing 97% CP-RS tubulin, 1% biotin brain tubulin, and 2% 647-Hilyte tubulin and paclitaxel-stabilized cisplatin resistant (CP-R) microtubules containing 99% CP-R tubulin, 1% biotin brain tubulin (Cytoskeleton, cat no. T333P) were immobilized on a glass surface using neutravidin.

Techniques: Cell-Signaling, Virus, Activation Assay, Control, Recombinant, Labeling, Mass Spectrometry, Sequencing, Cell Viability Assay, DNA Purification, Amplification, CRISPR, Plasmid Preparation, Synthesized, Software, Analysis, Filtration