18
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Screening of genes involved in epithelial‐mesenchymal transition and differential expression of complement‐related genes induced by PAX2 in renal tubules

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          ABSTRACT

          Aim

          The aim of the present study was to screen and verify downstream genes involved in the epithelial mesenchymal transition (EMT) induced by paired box 2 (PAX2) in NRK‐52E cells.

          Methods

          NRK‐52E cells were transfected with lentivirus carrying PAX2 gene or no‐load virus respectively. Total RNA was isolated 72 h after transfection from PAX2‐overexpressing cells and control cells. Isolated RNA was then hybridized with the Rat OneArray Plus expression profile chip. The chips were examined by Agilent 0.1 XDR to screen for differentially expressed genes, which were further analyzed to investigate complement‐related genes as genes of interest.

          Results

          In NRK‐52E cells, PAX2 overexpression promoted EMT followed by upregulation of 298 genes and downregulation of 293 genes. KEGG analysis indicated the differential expression of genes related to cytokines and their receptors, extracellular matrix (ECM), MAPKs, local adhesion, cancer, the complement cascade, and coagulation. Gene oncology analysis screened out genes related to molecular functions (e.g., hydrolase activity, phospholipase activity, components of the ECM) and biological processes (e.g., cell development, signal transduction, phylogeny), and cell components (e.g., cytoplasm, cell membrane, and ECM). Analysis of the complement system revealed upregulation of C3 and downregulation of CD55 and complement regulator factor H (CFH).

          Conclusion

          PAX2 overexpression upregulates EMT in vitro and may regulate C3, CD55, and CFH.

          Summary at a Glance

          This molecular analysis examines the effect of overexpressing paired box 2 (PAX2) in a tubule epithelial cell line. Results establish a link between pax2 and both epithelial‐mesenchymal transition (EMT) and the complement pathway.

          Related collections

          Most cited references26

          • Record: found
          • Abstract: found
          • Article: not found

          Local extravascular pool of C3 is a determinant of postischemic acute renal failure.

          The third complement component (C3) is an acute phase protein that plays a central role in reperfusion injury in several organ models. To investigate the contribution of local synthesis of C3 and distinguish it from that of circulating complement mainly produced by hepatic synthesis, we employed a mouse renal isograft model. Our model demonstrated a close relationship between the extent of intrarenal expression of C3 and cold-ischemia induced injury. Ischemic C3-positive donor kidneys transplanted into C3-positive or C3-negative recipients developed widespread tissue damage and severe acute renal failure. In contrast, ischemic C3-negative isografts exhibited only mild degrees of functional and structural disturbance, even when transplanted into normal C3-positive recipients. Thus local synthesis of C3, mostly identified in the tubular epithelium, was essential for complement-mediated reperfusion damage, whereas circulating C3 had a negligible effect. Our results suggest a two-compartment model for the pathogenic function of C3, in which the extravascular compartment is the domain of local synthesis of C3, and where the role of circulating C3 is redundant. Our data cast new light on the mechanism of complement-mediated tissue injury in nonimmunological disorders, and challenges the longstanding dogma that circulating components are the main complement effectors of extravascular tissue damage.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Endothelial-to-mesenchymal transition and renal fibrosis in ischaemia/reperfusion injury are mediated by complement anaphylatoxins and Akt pathway.

            Increasing evidence demonstrates a phenotypic plasticity of endothelial cells (ECs). Endothelial-to-mesenchymal transition (EndMT) contributes to the development of tissue fibrosis. However, the pathogenic factors and signalling pathways regulating this process in ischaemia/reperfusion (I/R) injury are still poorly understood. We investigated the possible role of complement in the induction of this endothelial dysfunction in a swine model of renal I/R injury by using recombinant C1 inhibitor in vivo. Here, we showed that I/R injury reduced the density of renal peritubular capillaries and induced tissue fibrosis with generation of CD31(+)/α-SMA(+) and CD31(+)/FPS-1(+) cells indicating EndMT. When we inhibited complement, the process of EndMT became rare, with preserved density of peritubular capillaries and significant reduction in renal fibrosis. When we activated ECs by anaphylatoxins in vitro, C3a and C5a led to altered endothelial phenotype with increased expression of fibroblast markers and decrease expression of specific endothelial markers. The activation of Akt pathway was pivotal for the C3a and C5a-induced EndMT in vitro. In accordance, inhibition of complement in vivo led to the abrogation of Akt signalling, with hampered EndMT and tissue fibrosis. Our data demonstrate a critical role for complement in the acute induction of EndMT via the Akt pathway. Therapeutic inhibition of these systems may be essential to prevent vascular damage and tissue fibrosis in transplanted kidney.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              C3a mediates epithelial-to-mesenchymal transition in proteinuric nephropathy.

              Tubulointerstitial inflammation and progressive fibrosis are common pathways that lead to kidney failure in proteinuric nephropathies. Activation of the complement system has been implicated in the development of tubulointerstitial injury in clinical and animal studies, but the mechanism by which complement induces kidney injury is not fully understood. Here, we studied the effect of complement on the phenotype of tubular epithelial cells. Tubular epithelial cells exposed to serum proteins adopted phenotypic and functional characteristics of mesenchymal cells. Expression of E-cadherin protein decreased and expression of both alpha-smooth muscle actin protein and collagen I mRNA increased. Exposure of the cells to the complement anaphylotoxin C3a induced similar features. Treating with a C3a receptor (C3aR) antagonist prevented both C3a- and serum-induced epithelial-to-mesenchymal transition. In the adriamycin-induced proteinuria model, C3aR-deficient mice demonstrated less injury, preserved renal function, and improved survival compared with wild-type mice. Furthermore, the kidneys of C3aR-deficient mice had significantly less interstitial collagen I and alpha-smooth muscle actin. In summary, the complement anaphylotoxin C3a is an important mediator of glomerular and tubulointerstitial injury and can induce tubular epithelial-to-mesenchymal transition.
                Bookmark

                Author and article information

                Contributors
                wuyb001@163.com
                Journal
                Nephrology (Carlton)
                Nephrology (Carlton)
                10.1111/(ISSN)1440-1797
                NEP
                Nephrology (Carlton, Vic.)
                John Wiley & Sons Australia, Ltd (Melbourne )
                1320-5358
                1440-1797
                24 March 2018
                February 2019
                : 24
                : 2 ( doiID: 10.1111/nep.2019.24.issue-2 )
                : 263-271
                Affiliations
                [ 1 ] Department of Pediatric Nephrology Shengjing Hospital of China Medical University Shenyang China
                Author notes
                [*] [* ] Correspondence:

                Professor Yu‐Bin Wu, Department of Pediatric Nephrology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, China. Email: wuyb001@ 123456163.com

                Article
                NEP13216
                10.1111/nep.13216
                6585862
                29280536
                91c2bd6e-1b2d-4198-833f-7829d435e6cc
                © 2017 The Authors Nephrology published by John Wiley & Sons Australia, Ltd on behalf of Asian Pacific Society of Nephrology

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

                History
                : 15 December 2017
                Page count
                Figures: 3, Tables: 5, Pages: 9, Words: 5070
                Funding
                Funded by: Science Plan Program of Liaoning Province, China
                Award ID: 2013021099
                Categories
                Original Article
                Original Articles
                Basic Research
                Custom metadata
                2.0
                nep13216
                February 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.4 mode:remove_FC converted:20.06.2019

                complement genes,epithelial‐mesenchymal cell transition,gene ontology,kyoto encyclopedia of genes and genomes,nrk‐52e cells,paired box 2

                Comments

                Comment on this article