Journal: Frontiers in Cell and Developmental Biology

Article Title: Network Pharmacology and Experimental Validation Reveal the Effects of Chidamide Combined With Aspirin on Acute Myeloid Leukemia-Myelodysplastic Syndrome Cells Through PI3K/AKT Pathway

doi: 10.3389/fcell.2021.685954

Figure Lengend Snippet: CDM combined with ASA inhibited the activation of PI3K/AKT pathway in AML-MDS cells. (A) Western blot analysis of PI3K, p-PI3K AKT, p-AKT. (B) Western blotting of PI3K, p-PI3K, AKT and p-AKT. (C) Western blotting of CDK2, CDK4 and p21. (D) Western blotting of Bcl-2, caspase-3 and cleaved caspase-3. IGF-1 reversed the effect of CDM combined with ASA on AML-MDS cells. β-actin served as a loading control. Data are mean ± SD of three independent experiments. “ ∗ ” indicates a significant difference relative to the control group ( ∗ p < 0.05), “&” indicates a significant difference relative to ASA-treated group (& p < 0.05), “NS” indicates no significant difference relative to the control group.

Article Snippet: PI3K (bsm-33219M), p-PI3K (AB1235888), and Caspase-3 (bs-0081R) from Bioss (Beijing, China).

Techniques: Activation Assay, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Network Pharmacology and Experimental Validation Reveal the Effects of Chidamide Combined With Aspirin on Acute Myeloid Leukemia-Myelodysplastic Syndrome Cells Through PI3K/AKT Pathway

doi: 10.3389/fcell.2021.685954

Figure Lengend Snippet: A schematic representation of the proposed pathway responsible for CDM combined with ASA in AML-MDS cells. (A) The predictive pathways of CDM and ASA in AML-MDS through network pharmacology. (B) CDM combined with ASA could inhibited the activation of PI3K/AKT signaling pathways, and then affected the expression of cell cycle and apoptosis-related proteins to induce cell cycle arrest and apoptosis in AML-MDS cells through experimental validation.

Article Snippet: PI3K (bsm-33219M), p-PI3K (AB1235888), and Caspase-3 (bs-0081R) from Bioss (Beijing, China).

Techniques: Activation Assay, Expressing

Journal: International Dental Journal

Article Title: High-Glucose Microenvironment Accelerates Malignant Progression Via O-GlcNAcylation in Oral Squamous Cell Carcinoma

doi: 10.1016/j.identj.2025.103897

Figure Lengend Snippet: The effects of OGT on cell proliferation, metastasis, and apoptosis partially depend upon the PI3K/AKT/mTOR signalling pathway in OSCC cells. (A-C) Levels of PI3K, AKT, mTOR, p-PI3K, p-AKT, p-mTOR, OGT, and O-GlcNAcylation proteins were assessed in OSCC cells exposed to a medium with different glucose concentrations and OGT knockdown. n = three independent experiments. Data were presented as the mean ± SD. P values were determined by one-way ANOVA. * P < .05, ** P < .01, *** P < .001, **** P < .0001.

Article Snippet: Antibodies against N-cadherin (N-cad) (1:1000 for WB, HY- P80238 ), E-cadherin (E-cad) (1:1000 for WB, P80113), vimentin (1:1000 for WB, HY- P80371 ), Bcl2 (1:1000 for WB, P80566 ), Bax (1:1000 for WB, P80028 ), PI3K (1:1000 for WB, HY- P80867 ), and phosphorylated P13K (p-PI3K) (1:1000 for WB, HY- P80846 ) were purchased from MedChemExpress; antibodies against AKT (1:1000 for WB, 4691T) and p-AKT (1:1000 for WB, 13038T) were purchased from Cell Signaling Technology; antibodies against OGT (1:5000 for WB, 1:250 for IHC, 66823-1-Ig), mammalian target of rapamycin (mTOR) (1:5000, 66888-1-Ig), p-mTOR (1:5000, 67778-1-Ig), and β-Actin (1:20,000 for WB, 66009-1-Ig) were obtained from Proteintech; and the O-GlcNAc antibody (1:1000 for WB, 1:200 for IHC, MA1-072) was purchased from Invitrogen.

Techniques: Knockdown

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Silencing MEG3 protects PC12 cells from hypoxic injury by targeting miR-21.

doi: 10.1080/21691401.2020.1725533

Figure Lengend Snippet: Figure 6. Silence of MEG3 activates PI3K/AKT pathway while deactivates NF-jB pathway via targeting miR-21. PC12 cells were co-transfected with si-MEG3/si-NC and miR-21 inhibitor/NC inhibitor, and then incubated in hypoxia for 24 h. The cells without transfection in normoxic condition served as control (CTRL). The expres- sion of proteins involved in (A,B) PI3K/AKT and (C,D) NF-jB pathways was tested by western blot. n ¼ 3.p < .05 represents a comparison of marked groups.

Article Snippet: Proteins were carried out by using primary antibodies specific for CyclinD1 (orb77046), p53 (orb178524), p21 (orb38089), Bcl-2 (orb378569), Bax (orb378566), pro-caspase-3 (orb13278), cleaved-caspase-3 (orb310032), PTEN (orb39110), PI3K (orb395443), p-PI3K (orb106105), AKT (orb29949), p-AKT (orb344403), IjBa (orb13487), p-IjBa (orb338946), p65 (orb344389), p-p65 (orb106211), and b-actin (orb378579, all from Biorbyt, San Francisco, CA).

Techniques: Transfection, Incubation, Control, Western Blot, Comparison

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Silencing MEG3 protects PC12 cells from hypoxic injury by targeting miR-21.

doi: 10.1080/21691401.2020.1725533

Figure Lengend Snippet: Figure 6. Silence of MEG3 activates PI3K/AKT pathway while deactivates NF-jB pathway via targeting miR-21. PC12 cells were co-transfected with si-MEG3/si-NC and miR-21 inhibitor/NC inhibitor, and then incubated in hypoxia for 24 h. The cells without transfection in normoxic condition served as control (CTRL). The expres- sion of proteins involved in (A,B) PI3K/AKT and (C,D) NF-jB pathways was tested by western blot. n ¼ 3.p < .05 represents a comparison of marked groups.

Article Snippet: Proteins were carried out by using primary antibodies specific for CyclinD1 (orb77046), p53 (orb178524), p21 (orb38089), Bcl-2 (orb378569), Bax (orb378566), pro-caspase-3 (orb13278), cleaved-caspase-3 (orb310032), PTEN (orb39110), PI3K (orb395443), p-PI3K (orb106105), AKT (orb29949), p-AKT (orb344403), IjBa (orb13487), p-IjBa (orb338946), p65 (orb344389), p-p65 (orb106211), and b-actin (orb378579, all from Biorbyt, San Francisco, CA).

Techniques: Transfection, Incubation, Control, Western Blot, Comparison

Journal: Journal of Oncology

Article Title: RGS20 Promotes Tumor Progression through Modulating PI3K/AKT Signaling Activation in Penile Cancer

doi: 10.1155/2022/1293622

Figure Lengend Snippet: RGS20 regulates PI3K/AKT signaling activation in PC cells. ( a) RNA-Seq analysis identified differentially expressed genes in Penl1 cells with or without RGS20 knockdown. (b) Top 5 enriched pathways for downregulated genes ( n = 118) following RGS20 knockdown in Penl1 cells. (c) GSEA analysis revealed that RGS20-high PC cases exhibited highly enriched PI3K/AKT signaling in GSE57955 dataset. (d) Coimmunoprecitation analysis on RGS20 interaction with PI3K p85 α subunit in PC cells. IgG was used as coimmunoprecitation control. (e) PI3K activity of RGS20-depleted 149RM and Penl1 cells. The PI3K activity in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001. (f) PI3K activity of 149RCa and LM156 cells with or without the RGS20 overexpression. The PI3K activity in EV control was regards as 100%. n = 4, ∗∗∗ P < 0.001. (g) PI3K activity of RGS20-depleted Penl1 cells after EGF stimulation. After serum deprivation for 36 hours, Penl1 cells (Scr or shRGS20) were stimulated with EGF (50 ng/mL) for 30 min, and then PI3K activity was analyzed. n = 4, ∗∗∗ P < 0.001. (h) Western blotting analysis on PI3K/AKT signaling proteins following RGS20 depletion in 149RM and Penl1 cells. (i) Western blotting analysis on PI3K/AKT signaling proteins following the overexpression of RGS20 in 149RCa and LM156 cells. (j) Western blotting analysis on PI3K/AKT signaling proteins in RGS20-depleted Penl1 cells after EGF stimulation. After serum deprivation for 36 hours, Penl1 cells (Scr or shRGS20) were stimulated with EGF (50 ng/mL) for 30 min. Western blotting was conducted to analyze PI3K/AKT signaling proteins.

Article Snippet: HA-tagged pCMV3 plasmid encoding empty vector (EV) or PI3K p110 α was purchased from Sinobiological (Beijing, China).

Techniques: Activation Assay, RNA Sequencing Assay, Activity Assay, Over Expression, Western Blot

Journal: Journal of Oncology

Article Title: RGS20 Promotes Tumor Progression through Modulating PI3K/AKT Signaling Activation in Penile Cancer

doi: 10.1155/2022/1293622

Figure Lengend Snippet: Knockdown of PI3K p85 α or p110 α suppresses cell proliferation, soft agar clonogenesis, and migration/invasion in PC cell lines. (a) Western blotting on PI3K p85 α or p110 α expression in 149RM and Penl1 cells transfected with scramble (Scr) control or shRNAs targeting p85 α or p110 α . β -Actin was used as loading control. (b) CCK-8 analysis on cell viability after p85 α or p110 α knockdown in 149RM and Penl1 cells. The cell viability in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001. (c) BrdU incorporation analysis on cell proliferation following p85 α or p110 α knockdown in 149RM and Penl1 cells. The BrdU incorporation in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001. (d) Soft agar clonogenesis of PC cells following p85 α or p110 α knockdown. The soft agar clonogenesis in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001. Bars: 100 μ m. (e) Caspase-3 activity of PC cells following p85 α or p110 α knockdown. The caspase-3 activity in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001. (f) Wound healing assay on PC cells following p85 α or p110 α knockdown. Micrographs showed the results of Penl1 cells. Bars: 100 μ m. The cell migration in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001. (g) Transwell invasion assay on PC cells following p85 α or p110 α knockdown. Bars: 50 μ m. The cell invasion in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001.

Article Snippet: HA-tagged pCMV3 plasmid encoding empty vector (EV) or PI3K p110 α was purchased from Sinobiological (Beijing, China).

Techniques: Migration, Western Blot, Expressing, Transfection, CCK-8 Assay, BrdU Incorporation Assay, Activity Assay, Wound Healing Assay, Transwell Invasion Assay

Journal: Journal of Oncology

Article Title: RGS20 Promotes Tumor Progression through Modulating PI3K/AKT Signaling Activation in Penile Cancer

doi: 10.1155/2022/1293622

Figure Lengend Snippet: The overexpression of constitutively active PI3K p110 α rescued malignant phenotypes impaired by RGS20 depletion. (a) Western blotting analysis on the expression of PI3K/AKT signaling pathway proteins after transfection with empty vector (EV), PI3K p110 α Myr, or PI3K p110 α KD plasmids in RGS20-depleted 149RM and Penl1cells. (b) CCK-8 analysis on cell viability after PI3K p110 α Myr and KD plasmids transfection in RGS20-depleted 149RM and Penl1 cells. The cell viability in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001, as compared with EV. (c) Soft agar assay on clonogenesis after PI3K p110 α Myr and KD plasmids transfection in RGS20-depleted 149RM and Penl1 cells. The soft agar clonogenesis in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001, as compared with EV. (d) Caspase-3 activity after PI3K p110 α Myr and KD plasmid transfection in RGS20-depleted 149RM and Penl1 cells. The caspase-3 activity in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001, as compared with EV. (e) Wound healing assay on PC cells following PI3K p110 α Myr and KD plasmid transfection in RGS20-depleted 149RM and Penl1 cells. Bars: 100 μ m. The cell migration in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001, as compared with EV. (f) Transwell invasion assay on PC cells following PI3K p110 α Myr and KD plasmid transfection in RGS20-depleted 149RM and Penl1 cells. Bars: 50 μ m. The cell invasion in Scr control was regards as 100%. n = 4, ∗∗∗ P < 0.001, as compared with EV.

Article Snippet: HA-tagged pCMV3 plasmid encoding empty vector (EV) or PI3K p110 α was purchased from Sinobiological (Beijing, China).

Techniques: Over Expression, Western Blot, Expressing, Transfection, Plasmid Preparation, CCK-8 Assay, Soft Agar Assay, Activity Assay, Wound Healing Assay, Migration, Transwell Invasion Assay

Journal: Journal of Oncology

Article Title: RGS20 Promotes Tumor Progression through Modulating PI3K/AKT Signaling Activation in Penile Cancer

doi: 10.1155/2022/1293622

Figure Lengend Snippet: RGS20 knockdown suppressed in vivo tumor growth and disrupted PI3K/AKT signaling activation in the Penl1 xenograft model . ( a). Nude mice xenograft study showed that RGS20 depletion attenuated subcutaneous tumor growth in nude mice. Tumor volume was measured every two days after Penl1 inoculation. n = 6, ∗ P < 0.05, shRGS20 vs. Scr control. (b) Tumor weight of Penl1 xenografts with or without RGS20 knockdown. ∗ P < 0.001. (c) Western blotting analysis on protein lysates extracted from Penl1 xenografts with or without RGS20 depletion ( n = 3). (d) IHC staining on Penl1 xenografts with or without RGS20 depletion at day 26 after inoculation. The tissue sections were incubated with antibodies against indicated antibodies (RGS20, p-AKT (T308), p-AKT (S473), Ki-67, and cleaved caspase-3). Bar, 50 μ m.

Article Snippet: HA-tagged pCMV3 plasmid encoding empty vector (EV) or PI3K p110 α was purchased from Sinobiological (Beijing, China).

Techniques: In Vivo, Activation Assay, Western Blot, Immunohistochemistry, Incubation

Journal: Journal of Oncology

Article Title: RGS20 Promotes Tumor Progression through Modulating PI3K/AKT Signaling Activation in Penile Cancer

doi: 10.1155/2022/1293622

Figure Lengend Snippet: The high RGS20 expression is associated with PI3K/AKT signaling activation and unfavorable patient survival in PC. (a) IHC micrographs showed consistent expression of RGS20 and p-AKT (T308, S473) in RGS20-high or RGS20-low PCs. Bars: 100 μ m. (b) The RGS20 expression was significantly correlated with p-AKT (T308) and p-AKT (S473) in our PC cohort ( n = 94). (c, d) Log-rank survival analysis showed that the high RGS20 expression was associated with progression-free survival and overall survival in our PC cohort ( n = 94).

Article Snippet: HA-tagged pCMV3 plasmid encoding empty vector (EV) or PI3K p110 α was purchased from Sinobiological (Beijing, China).

Techniques: Expressing, Activation Assay

Journal: PLOS ONE

Article Title: Distinctive genes and signaling pathways associated with type 2 diabetes-related periodontitis: Preliminary study

doi: 10.1371/journal.pone.0296925

Figure Lengend Snippet: The DEGs that are a part of the most critical canonical pathways in patients with T2DM and periodontitis compared with the patients with T2DM and without periodontitis.

Article Snippet: Afterward, the sections were incubated with primary antibodies against PTPN2 (1:500, polyclonal, rabbit, PA582516, ThermoFisher Scientific, Waltham, MA, USA), PTPN13 (1:500, polyclonal, rabbit, 25944-1-AP, Proteintech, Rosemont, IL, USA), DHCR24 (1:750, polyclonal, rabbit, 10471-1-AP), PIK3R2 (1:100, monoclonal, mouse, 67644-1-Ig, Proteintech, Rosemont, IL, USA), CALCRL (1:400, polyclonal, rabbit, PA550644, ThermoFisher Scientific, Waltham, MA, USA), IL1RN (1:2000, monoclonal, mouse, CF803396, ThermoFisher Scientific, Waltham, MA, USA), IL-6R alpha (1:350, polyclonal, rabbit, 23457-1-AP, Proteintech, Rosemont, IL, USA), and ITGA4 (1:500, polyclonal, rabbit, 10471-1-AP, ThermoFisher Scientific, Waltham, MA, USA) for 30 minutes.

Techniques: Protein Binding, Coagulation, Binding Assay, Activity Assay

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Design, Synthesis, and Biological Evaluation of Sulfonamide Methoxypyridine Derivatives as Novel PI3K/mTOR Dual Inhibitors.

doi: 10.3390/ph16030461

Figure Lengend Snippet: Figure 3. Compound 22c inhibited phosphorylation of PI3K downstream effectors AKT in HCT- 116 cells.

Article Snippet: To the kinase buffer, the compound solution, PI3K enzyme (SignalChem, Richmond, BC, Canada, catalog No. P27-122DH), PIP2 substrate, and ATP were diluted.

Techniques: Phospho-proteomics

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Design, Synthesis, and Biological Evaluation of Sulfonamide Methoxypyridine Derivatives as Novel PI3K/mTOR Dual Inhibitors.

doi: 10.3390/ph16030461

Figure Lengend Snippet: Figure 7. Predicted binding modes between 22c and the PI3Kα (left, PDB code for PI3Kα is 4JPS), mTOR (right, PDB code for mTOR is 4JT6). Hydrogen bonds are shown as yellow dashed lines. Images were generated with PyMOL.

Article Snippet: To the kinase buffer, the compound solution, PI3K enzyme (SignalChem, Richmond, BC, Canada, catalog No. P27-122DH), PIP2 substrate, and ATP were diluted.

Techniques: Binding Assay, Generated