Journal: Parasites & Vectors

Article Title: Ac -HSP20 regulates autophagy and promotes the encystation of Acanthamoeba castellanii by inhibiting the PI3K/AKT/mTOR signaling pathway

doi: 10.1186/s13071-024-06436-w

Figure Lengend Snippet: Effect of Ac -HSP20 on autophagy when the PI3K/AKT/mTOR pathway was inhibited and activated. Trophozoites were transfected with Ac -HSP20 siRNA, and encystation was induced for 48 h in an ice bath. Signal inhibitor: PI3K/mTOR inhibitor-11 (1 mmol/l). Signal activator: Recilisib (1.25 mmol/l). A Protein bands of LC3B and PI3K/AKT/mTOR pathway-related proteins after siRNA transfection were detected by western blot. B Statistical analysis on the protein expressions of LC3B and PI3K/AKT/mTOR pathway-related proteins. (The experiment was repeated three times. Compared to control group, * P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: PI3K/mTOR Inhibitor-11 (HY-151622) and Recilisib (HY-101625) were purchased from MCE (NJ, USA).

Techniques: Transfection, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: SERPIND1 Affects the Malignant Biological Behavior of Epithelial Ovarian Cancer via the PI3K/AKT Pathway: A Mechanistic Study

doi: 10.3389/fonc.2019.00954

Figure Lengend Snippet: SERPIND1 regulated the EMT of ovarian cancer cells via the PI3K/AKT pathway. (A) SERPIND1 overexpression in ES-2 cells resulted in a significant reduction in E-cadherin expression and increased expressions of N-cadherin, Vimentin, MMP2, MMP9, p-PI3K, and p-AKT. (B) Inhibition of SERPIND1 expression in CAOV3 cells resulted in increased E-cadherin expression and reduced expressions of N-cadherin, Vimentin, MMP2, MMP9, p-PI3K, and p-AKT. (C) Inhibition of SERPIND1 expression in OVCAR3 cells resulted in increased E-cadherin expression and reduced expressions of N-cadherin, Vimentin, MMP2, MMP9, p-PI3K, and p-AKT. SERPIND1, serpin family D member 1; PI3K, phosphoinositide 3-kinase; AKT, protein kinase B.

Article Snippet: To inhibit PI3K/AKT signaling, the cells were treated with a PI3K inhibitor, LY294002 (10 μM; Selleck, Houston, USA) for 48 h.

Techniques: Over Expression, Expressing, Inhibition

Journal: Frontiers in Oncology

Article Title: SERPIND1 Affects the Malignant Biological Behavior of Epithelial Ovarian Cancer via the PI3K/AKT Pathway: A Mechanistic Study

doi: 10.3389/fonc.2019.00954

Figure Lengend Snippet: The transcription factor NF-κB1 could regulate SERPIND1 expression, which in turn could activate the PI3K/AKT signaling pathway. (A) The PI3K/AKT pathway inhibitor LY294002 inhibited the promoting effect of SERPIND1 on the migration capacity of ovarian cancer cells. (B) LY294002 inhibited the promoting effect of SERPIND1 on the invasion capacity of ovarian cancer cells. (C) After the addition of LY294002, the SERPIND1-overexpressing ES-2 cells exhibited a reduced proportion of S-phase cells. (D) LY294002 inhibited the promoting effect of SERPIND1 on the proliferation capacity of ovarian cancer cells. (E) After the addition of LY294002, the SERPIND1-overexpressing ES-2 cells exhibited a significantly increased apoptotic rate. (F) A schematic diagram showing the proposed mechanism of NF-κB1 binding to the promoter region of SERPIND1. (G) NF-κB1 could bind to the promoter region of SERPIND1 at the −258 to −248 (GGTGTTTTCCA) and +131 to +141 (AGGCCTTCCTC) loci. (H) After inhibiting NF-κB1 expression, SERPIND1 expression was also reduced. Data are presented as mean ± standard deviation from three independent experiments. * P < 0.05. Scale bar: 100 μm. NF-κB1, nuclear factor kappa B1; SERPIND1, serpin family D member 1.

Article Snippet: To inhibit PI3K/AKT signaling, the cells were treated with a PI3K inhibitor, LY294002 (10 μM; Selleck, Houston, USA) for 48 h.

Techniques: Expressing, Migration, Binding Assay, Standard Deviation

Journal: Frontiers in Oncology

Article Title: SERPIND1 Affects the Malignant Biological Behavior of Epithelial Ovarian Cancer via the PI3K/AKT Pathway: A Mechanistic Study

doi: 10.3389/fonc.2019.00954

Figure Lengend Snippet: Schematic model showing: When the cells are subjected to external stimuli, NF-κB1 could enter the nucleus and exert its transcription factor function. The transcription factor NF-κB1 could bind to the promoter region of SERPIND1 and regulate SERPIND1 expression, which in turn could affect the PI3K/AKT pathway and promote the malignant biological behavior of ovarian cancer cells.

Article Snippet: To inhibit PI3K/AKT signaling, the cells were treated with a PI3K inhibitor, LY294002 (10 μM; Selleck, Houston, USA) for 48 h.

Techniques: 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: 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