Journal: Molecules

Article Title: The Potent G-Quadruplex-Binding Compound QN-302 Downregulates S100P Gene Expression in Cells and in an In Vivo Model of Pancreatic Cancer

doi: 10.3390/molecules28062452

Figure Lengend Snippet: ( A ) Structure of QN-302 (2,7-bis(3-morpholinopropyl)-4-((2-(pyrrolidin-1-yl)ethyl)amino)-9-(4-(pyrrolidin-1-ylmethyl)phenyl)benzo [lmn] [ , ]phenanthroline-1,3,6,8(2H,7H)-tetraone). ( B ) DNA sequence in the promoter region of the S100P gene found to form a G-quadruplex. The G4 sequence itself is bounded within the red box and the individual G-tracts are highlighted in red.

Article Snippet: Antibodies: Target Supplier Catalogue Number Dilution Incubation Details S100P Cell Signaling Technology 7677 1:1000 Overnight @4 °C GAPDH Cell Signaling Technology 5174 1:1000 Overnight @4 °C Anti-Rabbit IgG Cell Signaling Technology 7074 1:3000 RT for 1 h qPCR.

Techniques: Sequencing

Journal: Molecules

Article Title: The Potent G-Quadruplex-Binding Compound QN-302 Downregulates S100P Gene Expression in Cells and in an In Vivo Model of Pancreatic Cancer

doi: 10.3390/molecules28062452

Figure Lengend Snippet: Expression changes in selected genes previously identified as human PDAC-associated and responsive in QN-302 cell studies. Gene data taken from list of 229 genes with mutations and/or altered expression in PDAC, detailed in the Cancer Genetics Web site: http://www.cancerindex.org/geneweb/X0603.htm , accessed on 17 November 2022. mRNA data were taken from UCL RNA-seq transcriptome data on MIA PaCa-2 cells dosed with QN-302 [ 21 ]. PSQs are the number of putative quadruplexes found in a gene sequence.

Article Snippet: Antibodies: Target Supplier Catalogue Number Dilution Incubation Details S100P Cell Signaling Technology 7677 1:1000 Overnight @4 °C GAPDH Cell Signaling Technology 5174 1:1000 Overnight @4 °C Anti-Rabbit IgG Cell Signaling Technology 7074 1:3000 RT for 1 h qPCR.

Techniques: Expressing, Sequencing, Migration, Binding Assay, Transduction, Marker

Journal: Molecules

Article Title: The Potent G-Quadruplex-Binding Compound QN-302 Downregulates S100P Gene Expression in Cells and in an In Vivo Model of Pancreatic Cancer

doi: 10.3390/molecules28062452

Figure Lengend Snippet: ( A ) Western blots of S100P protein and GAPDH control, for protein extracted from the results of the xenograft experiment with MIA PaCa-2 implanted tumors and a vehicle control arm, at day 28 of the study. The number at the top of each individual column represents an individual mouse. QW: once-weekly dosing. BiW: twice-weekly dosing. ( B ) Quantitation of the Western blot data. Statistical significances are indicated by * p < 0.05, ** p < 0.01. ( C ) Quantitation of the qPCR data for the S100P gene in the treated vs. vehicle control animals. Statistical significances are indicated by * p < 0.05, ** p < 0.01 (one-way ANOVA test).

Article Snippet: Antibodies: Target Supplier Catalogue Number Dilution Incubation Details S100P Cell Signaling Technology 7677 1:1000 Overnight @4 °C GAPDH Cell Signaling Technology 5174 1:1000 Overnight @4 °C Anti-Rabbit IgG Cell Signaling Technology 7074 1:3000 RT for 1 h qPCR.

Techniques: Western Blot, Control, Quantitation Assay

Journal: Molecules

Article Title: The Potent G-Quadruplex-Binding Compound QN-302 Downregulates S100P Gene Expression in Cells and in an In Vivo Model of Pancreatic Cancer

doi: 10.3390/molecules28062452

Figure Lengend Snippet: Top predicted quadruplex sequences in the S100P gene, as found using the QGRS Mapper program ( https://bioinformatics.ramapo.edu/QGRS/index.php ), accessed on 12 June 2022. The G4 score, as defined in [ 43 ], is a measure of the likelihood of the sequence forming a stable quadruplex under physiological conditions. Only those sequences with a G-score >35 are listed here. G-tracts are underlined. The highlighted sequence, in the S100P promoter, is discussed further below.

Article Snippet: Antibodies: Target Supplier Catalogue Number Dilution Incubation Details S100P Cell Signaling Technology 7677 1:1000 Overnight @4 °C GAPDH Cell Signaling Technology 5174 1:1000 Overnight @4 °C Anti-Rabbit IgG Cell Signaling Technology 7074 1:3000 RT for 1 h qPCR.

Techniques: Sequencing

Journal: Molecules

Article Title: The Potent G-Quadruplex-Binding Compound QN-302 Downregulates S100P Gene Expression in Cells and in an In Vivo Model of Pancreatic Cancer

doi: 10.3390/molecules28062452

Figure Lengend Snippet: ( A ) Thermal difference spectra of 5 µM S100P PQS in 10 mM lithium cacodylate, 100 mM KCl buffer at pH 7.0. ( B ) CD spectra of 10 µM S100P quadruplex sequence ( B) in 10 mM lithium cacodylate, pH 7.0 and 100 mM of either KCl, NaCl or LiCl, as indicated. ( C ) Representative CD melting experiments with10 µM S100P quadruplex in 10 mM lithium cacodylate, pH 7.0, 100 mM KCl and 0, 10, 20 or 50 µM QN-302 as indicated.

Article Snippet: Antibodies: Target Supplier Catalogue Number Dilution Incubation Details S100P Cell Signaling Technology 7677 1:1000 Overnight @4 °C GAPDH Cell Signaling Technology 5174 1:1000 Overnight @4 °C Anti-Rabbit IgG Cell Signaling Technology 7074 1:3000 RT for 1 h qPCR.

Techniques: Circular Dichroism, Sequencing

Journal: Pharmaceuticals

Article Title: Jietacin Derivative Inhibits TNF-α-Mediated Inflammatory Cytokines Production via Suppression of the NF-κB Pathway in Synovial Cells

doi: 10.3390/ph16010005

Figure Lengend Snippet: Effect of jietacin derivative on the NF-κB pathway in SW982. Western blotting for p65, phosphorylated p65 (p-p65), importin α3, importin β1 and GAPDH ( A ). Densitometry of western blot protein bands for p-p65 ( B ), p65 ( C ), importin α3 ( D ), and importin β1 ( E ) were normalized to the expression of GAPDH ( n = 3). Vehicle, Lane 1,6,11; TNF-α, Lane 2,7,12; TNF-α+ 2.5 JD, Lane 3,8,13; TNF-α+ 1.25 JD, Lane 4,9,14; TNF-α+ 0.625 JD, Lane 5,10,15. a p < 0.05 in comparison with vehicle, b p < 0.05 compared to TNF-α.

Article Snippet: It was then incubated with anti-p65 mouse monoclonal antibody (1:10,000; cat no. #6956, Cell Signaling Technology, Boston, MA, USA), anti-phospho-p65 (Ser536) rabbit monoclonal antibody (1:10,000; cat no. #76778, Cell Signaling Technology), anti-importin α3 (affinity-purified rabbit polyclonal importin α3 antibody was raised against a synthetic peptide (C)GFNSSTNVPTEGFQF corresponding to the mouse importin α3 C-terminal region), anti-importin β1 rabbit monoclonal antibody (1:1000; cat no. #51186), anti-p38 MAPK (1:1000; cat no. #9212, Cell Signaling Technology), anti-phospho-p38 MAPK (Thr180/Tyr182) (1:1000; cat no. 9211, Cell Signaling Technology), or anti-GAPDH antibody (1:5000; FUJIFILM Wako Pure Chemical Co., Osaka, Japan) at 25 °C for 60 min.

Techniques: Western Blot, Expressing, Comparison

Journal: Pharmaceuticals

Article Title: Jietacin Derivative Inhibits TNF-α-Mediated Inflammatory Cytokines Production via Suppression of the NF-κB Pathway in Synovial Cells

doi: 10.3390/ph16010005

Figure Lengend Snippet: Effect of jietacin derivative on the NF-κB pathway in human primary synovial fibroblasts. Western blotting for p65, phosphorylated p65 (p-p65), importin α3, importin β1, and GAPDH ( A ). Densitometry of Western blot protein bands for p-p65 ( B ), p65 ( C ), importin α3 ( D ), and importin β1 ( E ) were normalized to the expression of GAPDH ( n = 3). Vehicle, Lane 1,4,7; TNF-α, Lane 2,5,8; TNF-α+ 2.5 JD Lane 3,6,9. a p < 0.05 compared with vehicle, b p < 0.05 compared to TNF-α.

Article Snippet: It was then incubated with anti-p65 mouse monoclonal antibody (1:10,000; cat no. #6956, Cell Signaling Technology, Boston, MA, USA), anti-phospho-p65 (Ser536) rabbit monoclonal antibody (1:10,000; cat no. #76778, Cell Signaling Technology), anti-importin α3 (affinity-purified rabbit polyclonal importin α3 antibody was raised against a synthetic peptide (C)GFNSSTNVPTEGFQF corresponding to the mouse importin α3 C-terminal region), anti-importin β1 rabbit monoclonal antibody (1:1000; cat no. #51186), anti-p38 MAPK (1:1000; cat no. #9212, Cell Signaling Technology), anti-phospho-p38 MAPK (Thr180/Tyr182) (1:1000; cat no. 9211, Cell Signaling Technology), or anti-GAPDH antibody (1:5000; FUJIFILM Wako Pure Chemical Co., Osaka, Japan) at 25 °C for 60 min.

Techniques: Western Blot, Expressing

Journal: Frontiers in Oncology

Article Title: CircRNA circ_0092314 Induces Epithelial-Mesenchymal Transition of Pancreatic Cancer Cells via Elevating the Expression of S100P by Sponging miR-671

doi: 10.3389/fonc.2021.675442

Figure Lengend Snippet: MiR-671 is downregulated in PAAD tissues and positively correlates with patient prognosis. (A) The predicted binding site of miR-671 within circ_0092314 and the S100P 3′-UTR sequence were shown. (B) The expression of miR-671 in PAAD tissues and adjacent normal tissues. (C) qRT-PCR analysis of miR-671 expression in four PAAD cell lines and normal pancreatic cells. (D) Kaplan-Meier curves for the overall survival of PAAD patients with high or low levels of miR-671 (KM Plotter database). ** P < 0.01, *** P < 0.001.

Article Snippet: Next, membranes were incubated with the primary antibodies: E-cadherin (Cell Signaling, MA, USA; 1:1000), Vimentin (Cell Signaling; 1:1000), S100P (Cell Signaling; 1:1000), AKT (Cell Signaling; 1:1000), p-AKT (Cell Signaling; 1:1000) and β-actin (Cell Signaling; 1:5000).

Techniques: Binding Assay, Sequencing, Expressing, Quantitative RT-PCR

Journal: Frontiers in Oncology

Article Title: CircRNA circ_0092314 Induces Epithelial-Mesenchymal Transition of Pancreatic Cancer Cells via Elevating the Expression of S100P by Sponging miR-671

doi: 10.3389/fonc.2021.675442

Figure Lengend Snippet: High S100P expression predicts poor prognosis in PAAD. (A) qRT-PCR analysis of S100P mRNA expression in PAAD tissues and adjacent normal tissues. (B) Immunohistochemical data was downloaded from the Human Protein Atlas database. The staining pattern for S100P protein in PAAD tissues and adjacent normal tissues were shown. (C) S100P mRNA expression in PAAD cells and HPDE6-C7 cells. (D) Kaplan-Meier curves for the overall survival of PAAD patients with high or low S100P expression (KM Plotter database). *** P < 0.001.

Article Snippet: Next, membranes were incubated with the primary antibodies: E-cadherin (Cell Signaling, MA, USA; 1:1000), Vimentin (Cell Signaling; 1:1000), S100P (Cell Signaling; 1:1000), AKT (Cell Signaling; 1:1000), p-AKT (Cell Signaling; 1:1000) and β-actin (Cell Signaling; 1:5000).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining, Staining

Journal: Frontiers in Oncology

Article Title: CircRNA circ_0092314 Induces Epithelial-Mesenchymal Transition of Pancreatic Cancer Cells via Elevating the Expression of S100P by Sponging miR-671

doi: 10.3389/fonc.2021.675442

Figure Lengend Snippet: MiR-671 suppresses EMT via targeting S100P. (A) The levels of S100P mRNA in PAAD cells transfected with miR-671 mimic or miR-671 inhibitor, respectively. (B) The activities of S100P 3′-UTR reporter containing WT or MUT miR-671 binding sites were determined using luciferase assay following co-transfection with miR-671 mimic or miR-671 inhibitor. (C) Western blotting analysis of the indicated proteins in PAAD cells transfected as indicated. (D, E) Cell invasion assay (left) and tumor sphere formation assay (right) in PaCa-2 cells transfected with (or without) miR-671 mimic, with (or without) S100P expression vector (D) , and in AsPC-1 cells transfected with (or without) miR-671 inhibitor, with (or without) S100P siRNA (E) . *** P < 0.001.

Article Snippet: Next, membranes were incubated with the primary antibodies: E-cadherin (Cell Signaling, MA, USA; 1:1000), Vimentin (Cell Signaling; 1:1000), S100P (Cell Signaling; 1:1000), AKT (Cell Signaling; 1:1000), p-AKT (Cell Signaling; 1:1000) and β-actin (Cell Signaling; 1:5000).

Techniques: Transfection, Binding Assay, Luciferase, Cotransfection, Western Blot, Invasion Assay, Tube Formation Assay, Expressing, Plasmid Preparation

Journal: Frontiers in Oncology

Article Title: CircRNA circ_0092314 Induces Epithelial-Mesenchymal Transition of Pancreatic Cancer Cells via Elevating the Expression of S100P by Sponging miR-671

doi: 10.3389/fonc.2021.675442

Figure Lengend Snippet: The correlation between circ_0092314 and miR-671/S100P expression in PAAD tissues. (A) The correlation between circ_0092314 expression and miR-671/S100P expression in PAAD tissues was examined using qRT-PCR assay. (B) A proposed mechanistic model in which circ_0092314 can sponge miR-671 to increase S100P expression, thereby promoting EMT and PAAD cell invasion.

Article Snippet: Next, membranes were incubated with the primary antibodies: E-cadherin (Cell Signaling, MA, USA; 1:1000), Vimentin (Cell Signaling; 1:1000), S100P (Cell Signaling; 1:1000), AKT (Cell Signaling; 1:1000), p-AKT (Cell Signaling; 1:1000) and β-actin (Cell Signaling; 1:5000).

Techniques: Expressing, Quantitative RT-PCR