Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM enhances EGFR resistance in lung cancer cells. (A) Dose‒effect relationship curves of osimertinib for PC-9 cells; (B) Dose‒effect relationship curves of osimertinib for PC-9 OR cells. (C) mRNA expression levels of ASPM and EGFR in PC-9 and PC-9 OR cells. (D) Protein expression of ASPM and EGFR in PC-9 and PC-9 OR cells. The histogram represents the relative gray values of the intracellular EGFR and ASPM proteins, and the data are presented as the means ± standard deviations (means ± SDs, n = 3 fields). (E) Silencing of the ASPM gene and mRNA expression levels of ASPM in PC-9 and PC-9 OR cells. (F) Dose‒effect relationship curves of osimertinib in PC-9 si ASPM cells. (G) Dose‒effect relationship curves of osimertinib in PC-9 OR si ASPM cells. (H) mRNA expression levels of ASPM were detected in PC-9 cells after overexpressing the ASPM gene. (I) Dose‒effect curves of osimertinib after PC-9 overexpression of ASPM . (J) The viability of the cells in each group was assessed via a CCK-8 assay. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing, Over Expression, CCK-8 Assay

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM plays a key role in the stabilization of drug-resistant cell lines. (A) Scatter plot of the correlation between ASPM and EGFR expression; p < 0.05. (B) Silencing of ASPM followed by ASPM and EGFR mRNA expression. (C) Protein expression of EGFR after silencing ASPM. (D) CHX administration at various time points. The right panel represents the relative gray values of the intracellular EGFR proteins, and the data are presented as the means ± standard deviations (means ± SDs, n = 3 fields). (E) IF images of PC-9 and PC-9 OR cells incubated with primary antibodies against EGFR (red) and ASPM (green). Detection was performed using secondary antibodies coupled to Alexa Fluor 488 and Alexa Fluor 555. The cell nuclei were stained with DAPI (blue), Scale = 7.5 μm. (F) mRNA expression levels of ASPM in PC-9 OR cells after silencing of ASPM -2. (G) PC-9 OR cells in the NC group/si ASPM -2 group were stained with propidium iodide (PI), and the DNA content was detected via flow cytometry. The red color represents the theoretical curve that we fit via ModFit software on the basis of the DNA content distribution data of the cells. (H) Statistics of the flow cytometric results in H plots. n = 3, a p value <0.05 was considered statistically significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing, Incubation, Staining, Flow Cytometry, Software

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM is significantly upregulated in NSCLC tumor tissues and strongly associated with reduced patient survival. ASPM silencing attenuates PC-9 and PC-9 OR malignant phenotypes, including proliferation and invasion, and sensitizes resistant cells to osimertinib. In addition, inhibiting the expression of ASPM effectively reduces damage to the cell cycle and protein stability of drug-resistant cells, thereby restoring the expression and function of EGFR.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing

Journal: The Eurasian Journal of Medicine

Article Title: Increased EGFR mRNA Expression Levels in Non-Small Cell Lung Cancer

doi: 10.5152/eurasianjmed.2016.0237

Figure Lengend Snippet: Primers for exon 21 PCR, exon 19 sequence analysis and expression of EGFR gene

Article Snippet: The immunohistochemical study was performed for protein expression and graded using Rabbit anti-human EGFR monoclonal (Clone SP9) antibody at a dilution of 1/100 (Spring Bioscience) according to the manufacturer’s instructions.

Techniques: Sequencing, Expressing

Journal: The Eurasian Journal of Medicine

Article Title: Increased EGFR mRNA Expression Levels in Non-Small Cell Lung Cancer

doi: 10.5152/eurasianjmed.2016.0237

Figure Lengend Snippet: Representative immunohistochemical staining intensity (left to right +1, +2, +3) for EGFR. Upper row: Adenocarcinoma, lower row: Squamous carcinoma

Article Snippet: The immunohistochemical study was performed for protein expression and graded using Rabbit anti-human EGFR monoclonal (Clone SP9) antibody at a dilution of 1/100 (Spring Bioscience) according to the manufacturer’s instructions.

Techniques: Immunohistochemical staining, Staining

Journal: The Eurasian Journal of Medicine

Article Title: Increased EGFR mRNA Expression Levels in Non-Small Cell Lung Cancer

doi: 10.5152/eurasianjmed.2016.0237

Figure Lengend Snippet: Characteristics of clinical patient according to EGFR mRNA and EGFR protein expression

Article Snippet: The immunohistochemical study was performed for protein expression and graded using Rabbit anti-human EGFR monoclonal (Clone SP9) antibody at a dilution of 1/100 (Spring Bioscience) according to the manufacturer’s instructions.

Techniques: Expressing

Journal: The Eurasian Journal of Medicine

Article Title: Increased EGFR mRNA Expression Levels in Non-Small Cell Lung Cancer

doi: 10.5152/eurasianjmed.2016.0237

Figure Lengend Snippet: Correlation between expression level of EGFR protein and mRNA

Article Snippet: The immunohistochemical study was performed for protein expression and graded using Rabbit anti-human EGFR monoclonal (Clone SP9) antibody at a dilution of 1/100 (Spring Bioscience) according to the manufacturer’s instructions.

Techniques: Expressing

Journal: The Eurasian Journal of Medicine

Article Title: Increased EGFR mRNA Expression Levels in Non-Small Cell Lung Cancer

doi: 10.5152/eurasianjmed.2016.0237

Figure Lengend Snippet: Summary of selected reports on EGFR gene, protein study in NSCLC

Article Snippet: The immunohistochemical study was performed for protein expression and graded using Rabbit anti-human EGFR monoclonal (Clone SP9) antibody at a dilution of 1/100 (Spring Bioscience) according to the manufacturer’s instructions.

Techniques: Mutagenesis, Fluorescence In Situ Hybridization, Real-time Polymerase Chain Reaction, Western Blot, Amplification, Quantitative RT-PCR, Expressing