Journal: Biomedical Reports

Article Title: Inhibition of SGK1 potentiates the anticancer activity of PI3K inhibitor in NSCLC cells through modulation of mTORC1, p‑ERK and β‑catenin signaling

doi: 10.3892/br.2023.1676

Figure Lengend Snippet: Combined inhibition of PI3K and SGK1 activity exerts a synergistic anticancer effect in non-small cell lung cancer cell lines. (A) IC 50 values of BYL719 alone or in combination with GSK650394 in CFA and CTG assay. (B) Dose-response curve of BYL719 alone and in combination with GSK650394 in (B) CTG (n=2) and (C) CFA (n=3). (D) Crystal violate stained colonies of NCI-H460 and A549 cells. (E) Bliss, Loewe, ZIP and HSA synergy scores for CFA and CTG assays. Combination scores >10 suggested synergism. SGK1, Serum and Glucocorticoid Kinase 1; IC 50 , Inhibitory Concentration 50%; ZIP, Zero Interaction Potency; HSA, Highest Single Agent; CFA, Colony Formation Assay; CTG, Cell TiterGlo; DRC, Dose Response Curve.

Article Snippet: PI3K inhibitor BYL719 was obtained from MedChemExpress (cat. no. HY-15244) and SGK1 inhibitor GSK650394 from Selleck Chemicals (cat. no. S7209).

Techniques: Inhibition, Activity Assay, CTG Assay, Staining, Concentration Assay, Colony Assay

Journal: Biomedical Reports

Article Title: Inhibition of SGK1 potentiates the anticancer activity of PI3K inhibitor in NSCLC cells through modulation of mTORC1, p‑ERK and β‑catenin signaling

doi: 10.3892/br.2023.1676

Figure Lengend Snippet: Combination effect of BYL719 and GSK650394 on the p-AKT/mTORC1/S6 pathway in non-small cell lung cancer cell lines. Western blot data showed combination regimen caused more robust inhibition of p-S6 (serine 240/244) and p-NDRG1 (threonine 346) levels than single agent treatment in (A) NCI-H460 cells and (B) A549 cells. p-AKT (serine 473) levels were inhibited following BYL719, but not GSK650394, treatment in both (A) NCI-H460 cells and (B) A549 cells. Densitometry analysis of p-AKT, p-NDRG1 and p-S6 levels in (C) NCI-H460 and (D) A549 cells confirmed inhibition of p-S6 by combination treatment. p-, phosphorylated; NDRG1, N-Myc downstream regulated 1.

Article Snippet: PI3K inhibitor BYL719 was obtained from MedChemExpress (cat. no. HY-15244) and SGK1 inhibitor GSK650394 from Selleck Chemicals (cat. no. S7209).

Techniques: Western Blot, Inhibition

Journal: Biomedical Reports

Article Title: Inhibition of SGK1 potentiates the anticancer activity of PI3K inhibitor in NSCLC cells through modulation of mTORC1, p‑ERK and β‑catenin signaling

doi: 10.3892/br.2023.1676

Figure Lengend Snippet: Combined inhibition of PI3K and SGK1 activity induces apoptosis, DNA damage and cell cycle arrest in NCI-H460 cells. (A) Western blot analysis showed enhanced proapoptotic (cleaved PARP1 and cleaved caspase 3) and decreased antiapoptotic markers (Bcl-xl). (B) Levels of the DNA damage marker (p-H2AX) increased following combination treatment. (C) Combination treatment significantly augmented BYL719-induced G0/G1 phase cell cycle arrest ( ** P<0.01, *** P<0.001), as observed by (D) flow cytometry analysis of PI staining. P2, G0/G1; P3, S; P4, G2/M; P5, sub-G1 phase. (E) Combination treatment modulated cyclin D1, CDK4, cyclin E1 and CDK2 levels, as determined using western blot. SGK1, Serum and Glucocorticoid kinase 1; p-, phosphorylated; H2AX, Histone 2A X; ns, non-significant.

Article Snippet: PI3K inhibitor BYL719 was obtained from MedChemExpress (cat. no. HY-15244) and SGK1 inhibitor GSK650394 from Selleck Chemicals (cat. no. S7209).

Techniques: Inhibition, Activity Assay, Western Blot, Marker, Flow Cytometry, Staining

Journal: Biomedical Reports

Article Title: Inhibition of SGK1 potentiates the anticancer activity of PI3K inhibitor in NSCLC cells through modulation of mTORC1, p‑ERK and β‑catenin signaling

doi: 10.3892/br.2023.1676

Figure Lengend Snippet: Combined effect of BYL719 and GSK650394 on the p-ERK/MAPK pathway in NSCLC cell lines. Combination regimen inhibited p-ERK, p-S6 (serine 235/236) and p-p90RSK (serine 380) levels in (A) NCI-H460 and (B) A549 cell line. Densitometry analysis of p-ERK and p-S6 levels in (C) NCI-H460 and (D) A549 cells confirmed higher inhibition of p-S6 (serine 235/236) levels in combination treatment. p-, phosphorylated; RSK, 90 kDa ribosomal S6 kinase.

Article Snippet: PI3K inhibitor BYL719 was obtained from MedChemExpress (cat. no. HY-15244) and SGK1 inhibitor GSK650394 from Selleck Chemicals (cat. no. S7209).

Techniques: Inhibition

Journal: Biomedical Reports

Article Title: Inhibition of SGK1 potentiates the anticancer activity of PI3K inhibitor in NSCLC cells through modulation of mTORC1, p‑ERK and β‑catenin signaling

doi: 10.3892/br.2023.1676

Figure Lengend Snippet: Combination of BYL719 and GSK650394 induces proteosomal degradation of β-catenin and its downstream targets c-Myc and c-Jun. Combination of BYL719 and GSK650394 resulted in a greater decrease of p-β-catenin, β-catenin, c-Myc and c-Jun levels than GSK650394-alone treatment in (A) NCI-H460 and (B) A549 cells. The combination treatment, but not either inhibitor alone, inhibited p-GSK3β levels. Densitometry analysis of β-catenin, c-Myc and c-Jun levels in (C) NCI-H460 and (D) A549 cells after 72 h treatment. (E) Co-treatment with MG-132 rescued β-catenin, c-Myc and c-Jun levels, confirming the proteosomal degradation of these proteins following combination treatment. p-, phosphorylated; GSK3β, Glycogen synthase kinase-3β.

Article Snippet: PI3K inhibitor BYL719 was obtained from MedChemExpress (cat. no. HY-15244) and SGK1 inhibitor GSK650394 from Selleck Chemicals (cat. no. S7209).

Techniques:

Journal: Virulence

Article Title: Neutrophil extracellular trap-borne C3-driven endothelial dysfunction in Klebsiella pneumoniae liver abscess

doi: 10.1080/21505594.2025.2580104

Figure Lengend Snippet: K. pneumoniae promotes net formation in neutrophils via the TLR/PI3K p110α/AKT pathway. (A) neutrophils were co-cultured with K. pneumoniae at different time points, and extracellular dna was detected using SYTOX green dye (scale bar = 100 μm). (B) Representative immunofluorescence images showing untreated neutrophils and neutrophils infected with K. pneumoniae ; dna (blue), mpo (red), and histone H3 (green); scale bar = 50 μm. (C) transmission electron microscopy showing the process of net release in neutrophils infected with K. pneumoniae . a. Untreated neutrophil with a clear lobulated nucleus (scale bar = 2 μm); b. Chromatin decondensation in the nucleus (scale bar = 2 μm); c. Vesicles containing dna chains fusing with the cell membrane and releasing into the extracellular space (scale bar = 1 μm); d. Vesicle rupture releasing NETs (scale bar = 2 μm). (D-E) Western blot analysis of PI3K-AKT signaling pathway proteins in untreated and K. pneumoniae -infected neutrophils, with β-actin as loading control; n = 3. (F-G) Western blot analysis of PI3K-AKT pathway proteins in neutrophils pretreated with TLR4 inhibitor before infection with K. pneumoniae , with β-actin as loading control; n = 3. (H) immunofluorescence staining of neutrophils pretreated with TLR4 inhibitor and PI3K p110α inhibitor after K. pneumoniae stimulation, showing dna (blue), mpo (red), and histone H3 (green); scale bar = 50 μm. (I-J). Western blot analysis and quantification of PAD4 protein in neutrophils pretreated with TLR4 inhibitor and PI3K p110α inhibitor after K. pneumoniae stimulation, with β-actin as loading control; n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: NETs were stained with 100 nM SYTOX Green (Cat: KGE2503-500, KeyGEN BioTECH, China), and the time-dependent induction of NETs was assessed from 0 to 60 min. To investigate the molecular mechanisms, neutrophils were co-incubated with K. pneumoniae and inhibitors, including TLR4 inhibitor (100 nM TAK-242, Cat: HY-11109, MedChemExpress, USA), PI3K p110α inhibitor (1 μM BYL-719, Cat: HY-15244, MedChemExpress, USA), DNase I (40 U, Cat: D8071, Solarbio, China), and C3 peptide inhibitor AMY-101 acetate (5 μM, Cat: HY-P1717B, MedChemExpress, USA).

Techniques: Cell Culture, Immunofluorescence, Infection, Transmission Assay, Electron Microscopy, Membrane, Western Blot, Control, Staining