Journal: iScience

Article Title: The potential therapeutic regimen for overcoming resistance to osimertinib due to rare mutations in NSCLC

doi: 10.1016/j.isci.2023.107105

Figure Lengend Snippet: PIK3CG and PIK3CA mutations induced osimertinib resistance in EGFR-mutant NSCLC cell lines or Ba/F3 cells (A) PIK3CG mutation increased PI3K (p110γ) and Akt/Bim signaling in Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M , HCC827, H3255, PC-9, and PC-9GR cells. Whole-cell protein lysates from mutant cells were subjected to immunoblotting to measure indicated proteins, with β-tubulin as a loading control. Similar results were obtained in three independent experiments. (B) Cell viability was analyzed by CCK-8 assay in PIK3CG mutant (−/+) Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M , HCC827, H3255, PC-9, and PC-9GR cells treated with osimertinib alone. Histograms show the IC50 of osimertinib in the indicated treatment groups. (C) PIK3CA mutation increased PI3K (p110α), and Akt/Bim signaling in Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M , HCC827, H3255, PC-9, and PC-9GR cells. Whole-cell protein lysates from mutant cells were subjected to immunoblotting to measure indicated proteins, with β-tubulin as a loading control. Similar results were obtained in three independent experiments. (D) Cell viability was analyzed by CCK-8 assay in PIK3CA mutant (−/+) Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M , HCC827, H3255, PC-9, and PC-9GR cells treated with osimertinib alone. Histograms show the IC50 of osimertinib in the indicated treatment groups. Numbers in the figures are mean values with S.D.

Article Snippet: H3255 , FineTest , C473.

Techniques: Mutagenesis, Western Blot, Control, CCK-8 Assay

Journal: iScience

Article Title: The potential therapeutic regimen for overcoming resistance to osimertinib due to rare mutations in NSCLC

doi: 10.1016/j.isci.2023.107105

Figure Lengend Snippet: Aspirin re-sensitized PIK3CG and PIK3CA mutant Ba/F3 and NSCLC cells to osimertinib in vitro (A) The effects of different doses of aspirin on various PI3KCG and PI3KCA mutant cell lines. Cell viabilities were examined 48 h after aspirin treatment. Assays were performed in triplicate. (B) Cell viability was analyzed by CCK-8 assay in PIK3CG and PIK3CA mutant (−/+) Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M , HCC827, H3255, PC-9, and PC-9GR cells treated with osimertinib alone or in combination with aspirin treatment for 48 h. Histograms show the IC50 of osimertinib in the indicated treatment groups. (C) Aspirin in combination with osimertinib increased apoptosis in PIK3CG and PIK3CA mutant Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M cells. Images are representative of three independent experiments. ∗p < 0.05; ∗∗p < 0.01. Asp: aspirin; Osi: osimertinib. Ctrl: control. (D) Aspirin decreased PI3K (p110γ), PI3K (p110α) and Akt/mTOR signaling, and activated Bim-related apoptosis signaling in PIK3CG and PIK3CA mutant HCC827, H3255, PC-9, and PC-9GR cells. Whole-cell protein lysates from PIK3CG and PIK3CA mutant cells with different treatments were subjected to immunoblotting to measure indicated proteins, with β-tubulin used as a loading control. Similar results were obtained in three independent experiments. Numbers in the figures are mean values with S.D.

Article Snippet: H3255 , FineTest , C473.

Techniques: Mutagenesis, In Vitro, CCK-8 Assay, Control, Western Blot

Journal: iScience

Article Title: The potential therapeutic regimen for overcoming resistance to osimertinib due to rare mutations in NSCLC

doi: 10.1016/j.isci.2023.107105

Figure Lengend Snippet: Aspirin sensitized osimertinib anti-tumor ability in PIK3CG mutant cell-derived xenograft mouse models (A) Macroscopic appearance of PC-9 PIK3CG -derived tumors at 4 weeks after drug administration. (B and C) Weight (g) and volume (mm 3 ) of PC-9 PIK3CG tumors treated with osimertinib, aspirin, and combined therapy. ∗∗, p < 0.01 when compared with the control; #, p < 0.05 as compared with osimertinib alone. (D) PC-9 PIK3CG nude mouse body weight was measured after indicated treatment. (E) The representative intensity images for each IHC score of PI3K (p110γ) and Ki67 staining in PC-9 PIK3CG tumor tissues are shown. Scale bars, 100 μm. Asp: aspirin; Osi: osimertinib; Ctrl: control. (F) Western blot showed aspirin decreased PI3K (p110γ) and Akt/mTOR signaling in PIK3CG and PIK3CA mutant Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M , HCC827, H3255, PC-9, and PC-9GR cells. Whole-cell protein lysates from PIK3CG mutant CDX tumors with different treatments were subjected to immunoblotting to measure indicated proteins, with β-tubulin used as a loading control. Numbers in the figures are mean values with SEM.

Article Snippet: H3255 , FineTest , C473.

Techniques: Mutagenesis, Derivative Assay, Control, Staining, Western Blot

Journal: iScience

Article Title: The potential therapeutic regimen for overcoming resistance to osimertinib due to rare mutations in NSCLC

doi: 10.1016/j.isci.2023.107105

Figure Lengend Snippet: Aspirin enhanced osimertinib anti-tumor ability in PIK3CA mutant patient-derived xenograft mouse models (A) Macroscopic appearance of PIK3CA mutant patient-derived tumors at 4 weeks after drug administration. (B and C) Weight (g) and volume (mm 3 ) of patient-derived tumors treated with osimertinib, aspirin, and combined therapy. ∗∗, p < 0.01 when compared with the control; #, p < 0.05 as compared with osimertinib alone. (D) The nude mouse body weight was measured after indicated treatment, and the data were shown as mean ± SEM (n = 3). (E) The representative intensity images for each IHC score of PI3K (p110α) and Ki67 staining in PIK3CA mutant patient-derived tumor tissues are shown. Scale bars, 100 μm. Asp: aspirin; Osi: osimertinib; Ctrl: control. Western blot showed aspirin decreased PI3K (p110α) and Akt/mTOR signaling in PIK3CG and PIK3CA mutant Ba/F3-EGFR Del19 , Ba/F3-EGFR Del19-T790M , HCC827, H3255, PC-9, and PC-9GR cells. (F) Whole-cell protein lysates from PIK3CA mutant PDX tumors with different treatments were subjected to immunoblotting to measure indicated proteins, with β-tubulin used as a loading control. Numbers in the figures are mean values with SEM.

Article Snippet: H3255 , FineTest , C473.

Techniques: Mutagenesis, Derivative Assay, Control, Staining, Western Blot

Journal: iScience

Article Title: The potential therapeutic regimen for overcoming resistance to osimertinib due to rare mutations in NSCLC

doi: 10.1016/j.isci.2023.107105

Figure Lengend Snippet:

Article Snippet: H3255 , FineTest , C473.

Techniques: Virus, Recombinant, CCK-8 Assay, Bicinchoninic Acid Protein Assay, Software