Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Simplified schematic illustration of pathways for Wnt/β-catenin, ERK/MAPK and PI3K/Akt and PGE 2 -metabolism. A) Canonical Wnt/β-catenin signaling. The engagement of the Wnt receptor, Frizzled, leads to the inhibition of the β-catenin destruction complex, composed of APC, axin and GSK3β. β-catenin thereby avoids ubiquitination and subsequent degradation, thus allowing it to translocate to the nucleus to activate an array of regulatory genes. B) The RAS/RAF/MEK/ERK MAPK pathway. Stimulation of the receptor tyrosine kinase (RTK) or G-protein coupled receptors (GPCRs) leads to sequential activation of RAS, RAF, MEK, and ERK causing modification of substrates promoting cell survival and proliferation. C) In the PI3K/Akt pathway, activation of the RTK or GPCRs leads to sequential modification of phosphatidyl inositol residues of the phospholipid bilayer. In this process, PI3K generates PIP3. PIP3 in association with PDK1 activates Akt. Akt then modulates the activity of downstream substrates including mTOR, thus promoting proliferation and cell survival. D) PGE 2 -metabolism. PGE 2 -synthesis begins with catalytic hydrolysis of membrane phospholipids by cytoplasmic phospholipase A2 (cPLA 2 ), thus releasing arachidonic acid (AA). By the action of the COX-1 and COX-2, AA is converted to prostaglandin H 2 (PGH 2 ). PGH 2 is then converted to PGE 2 by prostaglandin E synthase (PTGES). The main exporter of PGE 2 is thought to be multi-drug resistance related polypeptide 4 (MRP4). Removal of PGE 2 from the extracellular compartment around target cells occurs by diffusion to the blood stream and subsequent uptake and degradation in lung, liver or kidney endothelial cells or by import to colonic epithelial cells through the prostaglandin transporter (PGT) and subsequent degradation by 15-prostaglandin dehydrogenase (15-PGDH). Through autocrine and paracrine signaling, extracellular PGE 2 stimulates the prostaglandin receptors EP1–4. The EPs are GPCRs with EP1 being Gαq-coupled while EP2 and EP4 are Gαs-coupled. EP3 is capable of coupling with different G-proteins including Gαi, Gαs and Gαq

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Inhibition, Activation Assay, Modification, Activity Assay, Diffusion-based Assay

Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Quantitative real-time PCR primer sequences

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Real-time Polymerase Chain Reaction

Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Difference in mRNA expression in CRN versus controls

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Expressing

Journal: Cancers

Article Title: Genome-Wide Comparison of the Target Genes of the Reactive Oxygen Species and Non-Reactive Oxygen Species Constituents of Cold Atmospheric Plasma in Cancer Cells

doi: 10.3390/cancers12092640

Figure Lengend Snippet: Pathway analysis of genes deregulated by H 2 O 2 and CAP. ( A ) KEGG pathway analysis of genes regulated by H 2 O 2 or CAP. Pathways observed in both U937 and SK-mel-147 cells are shown. The larger the size of the circle, the more genes included in the pathway. The ‘MAPK pathway’ and ‘Pathways in cancer’, which include PTGER3 and HSPA6 from CAP-treated cells, are indicated with an asterisk (*). ( B ) Venn diagram of common genes regulated in U937 and SK-mel-147 cells by H 2 O 2 or CAP. There were four and two common genes significantly altered by H 2 O 2 and CAP, respectively, in both cells. ( C ) Expression profile of PTGER3 and HSPA6 in various tumors. The expression level of the genes was extracted from the CCLE database and aligned with the box plot. The horizontal line indicates the expression level of control normal tissues. The numeric value in the parenthesis represents the number of examined tissues.

Article Snippet: The antibodies used were anti-HSPA6 (1:1000, NBP1-32761; Novus Biologicals, Centennial, CO, USA), anti-PTGER3 (1:1000, PAB25115; Abnova, Taipei, Taiwan), and anti-β-actin (1:1000, bs-0061R; Bioss, Woburn, MA, USA).

Techniques: Expressing, Control

Journal: Cancers

Article Title: Genome-Wide Comparison of the Target Genes of the Reactive Oxygen Species and Non-Reactive Oxygen Species Constituents of Cold Atmospheric Plasma in Cancer Cells

doi: 10.3390/cancers12092640

Figure Lengend Snippet: Regulation of PTGER3 and HSPA6 by the non-ROS components of CAP at the protein level. Western blot analysis was performed for PTGER3 and HSPA6 after treating U937 cells with H 2 O 2 or CAP in the presence of NAC. ( A ) PTGER3 after H 2 O 2 and CAP treatment with NAC. ( B ) HSPA6 after H 2 O 2 and CAP treatment with NAC. All assays were performed in triplicate, and the results are presented as the mean ± SE. *** p < 0.001; ns: Non-significant.

Article Snippet: The antibodies used were anti-HSPA6 (1:1000, NBP1-32761; Novus Biologicals, Centennial, CO, USA), anti-PTGER3 (1:1000, PAB25115; Abnova, Taipei, Taiwan), and anti-β-actin (1:1000, bs-0061R; Bioss, Woburn, MA, USA).

Techniques: Western Blot

Journal: Cancers

Article Title: Genome-Wide Comparison of the Target Genes of the Reactive Oxygen Species and Non-Reactive Oxygen Species Constituents of Cold Atmospheric Plasma in Cancer Cells

doi: 10.3390/cancers12092640

Figure Lengend Snippet: Suppression of U937 cancer cell growth by siRNAs for PTGER3 and HSPA6. PTGER3 and HSPA6 were downregulated in U937 cells by transient transfection with siRNAs, and the effect on cell proliferation and apoptosis was assessed by ( A ) CCK-8 assay and ( B , C ) flow cytometry analysis, respectively. siNC: Negative control siRNA; siPTGER3: siRNA for PTGER3; siHSPA6: siRNA for HSPA6. Representative images are shown for the flow cytometry analysis. All experiments were performed at least three times, and the results are presented as the mean ± SE. * p < 0.05; ** p < 0.01; ns: Non-significant.

Article Snippet: The antibodies used were anti-HSPA6 (1:1000, NBP1-32761; Novus Biologicals, Centennial, CO, USA), anti-PTGER3 (1:1000, PAB25115; Abnova, Taipei, Taiwan), and anti-β-actin (1:1000, bs-0061R; Bioss, Woburn, MA, USA).

Techniques: Transfection, CCK-8 Assay, Flow Cytometry, Negative Control