Journal: Journal of Pharmaceutical Analysis

Article Title: Synergistic effects of methyl 2-cyano-3,11-dioxo-18beta-olean-1,-12-dien-30-oate and erlotinib on erlotinib-resistant non-small cell lung cancer cells

doi: 10.1016/j.jpha.2021.06.002

Figure Lengend Snippet: Cell viability summary showing calculated IC 50 values following 48 h treatment with various tyrosine kinase inhibitors in both HCC827 and HCC827R cells.

Article Snippet: ERL, tivantinib, dasatinib, osimertinib, olmutinib, crizotinib, and cabozantinib were purchased from MedChemExpress LLC (Princeton, NJ, USA).

Techniques:

Journal: Journal of Pharmaceutical Analysis

Article Title: Synergistic effects of methyl 2-cyano-3,11-dioxo-18beta-olean-1,-12-dien-30-oate and erlotinib on erlotinib-resistant non-small cell lung cancer cells

doi: 10.1016/j.jpha.2021.06.002

Figure Lengend Snippet: Cell viability summary showing calculated IC 50 values following simultaneous 48 h treatment with various tyrosine kinase inhibitors in combination with a constant concentration of 2 μM CDODA-Me in both HCC827 and HCC827R cells.

Article Snippet: ERL, tivantinib, dasatinib, osimertinib, olmutinib, crizotinib, and cabozantinib were purchased from MedChemExpress LLC (Princeton, NJ, USA).

Techniques: Concentration Assay

Journal: F1000Research

Article Title: Ex-vivo drug screening of surgically resected glioma stem cells to replace murine avatars and provide personalise cancer therapy for glioblastoma patients

doi: 10.12688/f1000research.135809.1

Figure Lengend Snippet: Drug compounds and concentrations used on 384 well drug plate.

Article Snippet: AZD9291 (Osimertinib) , Stratech (Selleckchem) , 5 , 2.5 , 1.25 , 0.625.

Techniques: Concentration Assay

Journal: Acta Biochimica et Biophysica Sinica

Article Title: CCL2 promotes EGFR-TKIs resistance in non-small cell lung cancer via the AKT-EMT pathway

doi: 10.3724/abbs.2024106

Figure Lengend Snippet: CCL2 is upregulated in NSCLC cells with acquired resistance to EGFR-TKIs (A) Clustered heatmap of the upregulated ( P-value<0.05 and log2FC≥1) and downregulated genes ( P-value<0.05 and log2FC<1) between gefitinib-resistant (gefi-R) and gefitinib-sensitive (gefi-S) NSCLC cells. (B) KEGG pathway analysis of gefitinib-resistant and gefitinib-sensitive NSCLC samples in the GSE122005 dataset. (C) Volcano plot showing genes that were differentially expressed between gefitinib-resistant and gefitinib-sensitive NSCLC cells. (D,E) Cells were treated with different concentrations of gefitinib or osimertinib for 72 h. CCK-8 assays were used to detect the IC 50 values of gefitinib in HCC827 and HCC827-GR cells (D) and the IC 50 values of osimertinib in HCC827 and HCC827-OR cells (E). (F) qRT-PCR was used to assess the mRNA expression of CCL2 in HCC827, HCC827-GR, and HCC827-OR cells. (G) ELISA was performed to detect the protein expressions of CCL2 in HCC827, HCC827-GR, and HCC827-OR cells. Data are shown as the mean±SEM of three independent experiments (*** P<0.001).

Article Snippet: Osimertinib (AZD-9291, GC16308) and gefitinib (IG0060) were procured from GLPBIO (Shanghai, China) and Solarbio (Beijing, China), respectively.

Techniques: CCK-8 Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Acta Biochimica et Biophysica Sinica

Article Title: CCL2 promotes EGFR-TKIs resistance in non-small cell lung cancer via the AKT-EMT pathway

doi: 10.3724/abbs.2024106

Figure Lengend Snippet: CCL2 upregulation induces resistance to EGFR-TKIs in NSCLC cells (A,B) The viability of HCC827 cells treated with gefitinib (A) and osimertinib (B) was determined by a CCK-8 assay after pretreatment with rhCCL2 (100 ng/mL) or PBS (control) for 120 h. (C‒E) Validation of CCL2 overexpression in HCC827 cells at the mRNA (D) and protein (C,E) levels. (F,G) The viability of gefitinib-(F) and osimertinib-(G) treated HCC827 cells after CCL2 overexpression was determined by CCK-8 assay. Data are shown as the mean±SEM of three independent experiments (** P<0.01, *** P<0.001).

Article Snippet: Osimertinib (AZD-9291, GC16308) and gefitinib (IG0060) were procured from GLPBIO (Shanghai, China) and Solarbio (Beijing, China), respectively.

Techniques: CCK-8 Assay, Control, Biomarker Discovery, Over Expression

Journal: Acta Biochimica et Biophysica Sinica

Article Title: CCL2 promotes EGFR-TKIs resistance in non-small cell lung cancer via the AKT-EMT pathway

doi: 10.3724/abbs.2024106

Figure Lengend Snippet: CCL2 suppression reverses EGFR-TKIs resistance in vitro (A,B) In the presence or absence of bindarit (300 nM), HCC827 cells were pretreated with rhCCL2 (100 ng/mL) for 120 h, and the sensitivities to gefitinib (A) and osimertinib (B) were tested by CCK-8 assay. (C,D) Viability of HCC827-GR cells treated with gefitinib (C) and HCC827-OR cells treated with osimertinib (D) pretreated with or without bindarit (300 nM) was assayed by CCK-8 assay. (E,F) Validation of CCL2 knockdown in HCC827-GR cells and HCC827-OR cells at the mRNA (E) and protein levels (F). (G,H) The viability of HCC827-GR cells treated with gefitinib (G) and HCC827-OR cells treated with osimertinib (H) was determined by CCK-8 assay after CCL2 knockdown. Data are shown as the mean±SEM of three independent experiments (* P<0.05, ** P<0.01, *** P<0.001).

Article Snippet: Osimertinib (AZD-9291, GC16308) and gefitinib (IG0060) were procured from GLPBIO (Shanghai, China) and Solarbio (Beijing, China), respectively.

Techniques: In Vitro, CCK-8 Assay, Biomarker Discovery, Knockdown

Journal: Cell Communication and Signaling : CCS

Article Title: Dual blockade of EGFR and PI3K signaling pathways offers a therapeutic strategy for glioblastoma

doi: 10.1186/s12964-023-01400-0

Figure Lengend Snippet: AZD-9291 and GDC-0084 induce cell cycle arrest in G0/G1 phase. A The differentially expressed genes (DEGs) were analyzed by transcriptome sequencing after combination treatment. According to the volcano scatter plot of expressed genes, 2021 genes were up-regulated and 2037 genes were down-regulated after AZD-9291 and GDC-0084 combination treatment. |Log2FC| > 1 & p.adj < 0.05. KEGG pathway ( B ) and Gene set enrichment analysis (GSEA) ( C ) enrichment analysis of the DEGs in AZD-9291 and GDC-0084 co-treated cells vs. control cells. D Cell cycle analyses of LN229 and U251 treated by AZD-9291 (2 μΜ) and/or GDC-0084 (2 μΜ) by flow cytometry. E , F LN229 and U251 cells were treated with AZD-9291 (2 μΜ), GDC-0084 (2 μΜ) or combination for 24 h. Cell lysates were analyzed for Cyclin D1 and p21 by western blot analysis

Article Snippet: EGFR inhibitor AZD-9291 and dual PI3K/mTOR inhibitor GDC-0084 were obtained from CSNpharm (CSNpharm, Chicago, IL, US).

Techniques: Sequencing, Flow Cytometry, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Dual blockade of EGFR and PI3K signaling pathways offers a therapeutic strategy for glioblastoma

doi: 10.1186/s12964-023-01400-0

Figure Lengend Snippet: The combination of AZD-9291 and GDC-0084 can simultaneously block EGFR/MEK/ERK and PI3K/AKT/mTOR signaling pathways. A DEGs in the control, GDC-0084, AZD-9291 and AZD + GDC treatment groups with triplicates are shown in the heat map. Gradient color barcode indicated fold change of expression (Log2). B GSEA was used to analyze the signaling pathways enrichment in different groups. C , D Representative western blot analysis showing the effects of AZD-9291 combined with GDC-0084 on PI3K/AKT/mTOR and EGFR/MEK/ERK signaling pathways in LN229 and U251 cells. The expression levels of core protein of these two signaling pathways were examined by using indicated antibodies

Article Snippet: EGFR inhibitor AZD-9291 and dual PI3K/mTOR inhibitor GDC-0084 were obtained from CSNpharm (CSNpharm, Chicago, IL, US).

Techniques: Blocking Assay, Expressing, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Dual blockade of EGFR and PI3K signaling pathways offers a therapeutic strategy for glioblastoma

doi: 10.1186/s12964-023-01400-0

Figure Lengend Snippet: AZD-9291 combined with GDC-0084 synergistically decreases the survival and inhibits colony formation of primary GBM cells. A Three primary GBM cell lines were treated with AZD-9291 alone (1 μΜ), GDC-0084 alone (0.5 μΜ) or their combination for 72 h. Cell viability was then measured by CCK-8 assay. B , C Representative images of the EdU incorporation assay and Quantitative analysis of the results, scale bar: 100 μm. D , E Colony formation ability of GBM2 and GBM3 cells following AZD-9291 (1 μΜ) and/or GDC-0084 (0.5 μΜ). The numbers of colony formation were normalized to the control group. F GBM2 and GBM3 cells were exposed to 0.1%DMSO, 1 μΜ AZD-9291 alone, 0.5 μM GDC-0084 alone and AZD-9291 combined with GDC-0084 for 24 h. The expression levels of EGFR, p-EGFR, ERK1/2, p-ERK1/2, AKT and p-AKT were evaluated by Western blotting. GAPDH was used as loading control. All the Data are presented as means ± SD. **, P < 0.01, ***, P < 0.001

Article Snippet: EGFR inhibitor AZD-9291 and dual PI3K/mTOR inhibitor GDC-0084 were obtained from CSNpharm (CSNpharm, Chicago, IL, US).

Techniques: CCK-8 Assay, Expressing, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Dual blockade of EGFR and PI3K signaling pathways offers a therapeutic strategy for glioblastoma

doi: 10.1186/s12964-023-01400-0

Figure Lengend Snippet: Combining AZD-9291 with GDC-0084 attenuated the growth of glioma in vivo. A Representative tumors isolated from the control, GDC-0084 (15 mg/kg), AZD-9291 (15 mg/kg) and AZD + GDC-treated groups of subcutaneous tumor model. B Tumor volume were recorded every 3 days. C Tumors isolated from each treatment group were weighted. The tumor weight was analyzed statistically. D Whole-tumor protein lysates were prepared from three randomly chosen tumors in each group to detect the levels of p-AKT and p-ERK1/2 using western blot analysis in vivo. E Schematic representation of the LN229-derived orthotopic xenograft experimental workflow. F Representative images of H&E staining of whole-brain sections from groups with different drugs administration. G Representative bioluminescence images of intracranial xenografts of each group on the indicated days after implantation. H Quantitative analysis of the results in ( G ). I Kaplan-Meier survival curves of mice implanted with LN229 cells with different drugs administration. A log-rank test was used to assess the statistical significance of the differences ( n = 6). J Representative IHC staining images of Ki67 expression in LN-229-derived xenograft tumor of each group. Scale bar: 20 μm. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: EGFR inhibitor AZD-9291 and dual PI3K/mTOR inhibitor GDC-0084 were obtained from CSNpharm (CSNpharm, Chicago, IL, US).

Techniques: In Vivo, Isolation, Western Blot, Derivative Assay, Staining, Immunohistochemistry, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Dual blockade of EGFR and PI3K signaling pathways offers a therapeutic strategy for glioblastoma

doi: 10.1186/s12964-023-01400-0

Figure Lengend Snippet: Combinatorial treatment with AZD-9291 and GDC-0084 synergistically inhibits cell viability and colony formation in GBM cells. A - D Increasing concentrations of AZD-9291, GDC-0084 or both were used to treat four GBM cell lines for 72 h. Cell viability was then measured by CCK-8 assay. E , F LN229 and U251 cells were treated with AZD-9291 (2 μΜ) and/or GDC-0084 (2 μΜ) for 24 h, and then changed with drug-free medium for another 14 days. The numbers of colony formation were counted. F Quantitative analysis of the results in ( E ). The numbers of colony formation were normalized to the control group. G , H Representative images ( H ) and quantitative results ( G ) of EdU assay in LN229 and U251 cells treated with AZD-9291 (2 μΜ) and/or GDC-0084 (2 μΜ), scale bar: 100 μm. All the data were presented as means ± SD from three independent experiments (* P < 0.05, ** P < 0.01)

Article Snippet: EGFR inhibitor AZD-9291 and dual PI3K/mTOR inhibitor GDC-0084 were obtained from CSNpharm (CSNpharm, Chicago, IL, US).

Techniques: CCK-8 Assay, EdU Assay