Knockdown of mediator subunit Med19 suppresses bladder cancer cell proliferation and migration by downregulating Wnt/β-catenin signalling pathway
Authors: Zhenli Gao
Diandong Yang
Hejia Yuan
Zhe Zhu
Jitao Wu
Yuanshan Cui
Changping Men
Shengqiang Yu
Journal: Journal of Cellular and Molecular Medicine (2017)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Med19; Wnt/β-catenin pathway; bladder cancer; migration; proliferation.
Affiliations: Department of Urology, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong, China., Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA.

Abstract:  Mediator complex subunit 19 (Med19), a RNA polymerase II-embedded coactivator, is reported to be involved in bladder cancer (BCa) progression, but its functional contribution to this process is poorly understood. Here, we investigate the effects of Med19 on malignant behaviours of BCa, as well as to elucidate the possible mechanisms. Med19 expression in 15 BCa tissues was significantly higher than adjacent paired normal tissues using real-time PCR and Western blot analysis. Immunohistochemical staining of 167 paraffin-embedded BCa tissues was performed, and the results showed that high Med19 protein level was positively correlated with clinical stages and histopathological grade. Med19 was knocked down in BCa cells using short-hairpin RNA. Functional assays showed that knocking-down of Med19 can suppress cell proliferation and migration in T24, UM-UC3 cells and 5637 in vitro, and inhibited BCa tumour growth in vivo. TOP/FOPflash reporter assay revealed that Med19 knockdown decreased the activity of Wnt/β-catenin pathway, and the target genes of Wnt/β-catenin pathway were down-regulated, including Wnt2, β-catenin, Cyclin-D1 and MMP-9. However, protein levels of Gsk3β and E-cadherin were elevated. Our data suggest that Med19 expression correlates with aggressive characteristics of BCa and Med19 knockdown suppresses the proliferation and migration of BCa cells through down-regulating the Wnt/β-catenin pathway, thereby highlighting Med19 as a potential therapeutic target for BCa treatment. 

Retracted: No

PubPeer comments:
 Figure 3A 5637 cells. There seems a partial duplication after vertical flip ![file] This work was supported by grants from the National Nature Science Foundation of China (Nos. 81572835; 81302234), Nature Science Foundation of Shandong Province (No. ZR2015HM021). , 
 Fig 5A.  Inhibition of BCa cell proliferation and migration by Med19 knockdown can be rescued by LiCl. (A) BCa cells were treated with or without 10 mM LiCl, the protein expression of u03b2u2010catenin was examined by Western blot. ![file] There seems to be an error in the loading control bands. , 
 An image from Figure 5D of this paper seems to overlap (after rotation) with an image in Figure 4B April 2017, Oncotarget, DOI:10.18632 oncotarget.17021, discussed here: https: pubpeer.com publications 790FFBC9A029E5E1E345EBE5ABDE9B Because this image only partially overlaps with its cousin, it seems that whomever prepared this figure had access to the original source. *** ![file] , 
 Thank you for raising this concern, we are so sorry to find that the wound healing pictures of the bladder cancer cells were chosen by mistake. The loading control bands were mismatched in Fig. 5A. As for the overlap of the image between two papers, these two articles came from the same center, close in time, cross membership and similar experiments. We work as a multidisciplinary team, always learning, collaborating and debating. The picture of Oncotarget was chosen by mistake. We have reached the editorial office of JCMM and Oncotarget for mistakes correction. We ll keep you informed of the progress made. Yours sincerely, Hejia Yuan , 
 #4 Thank you for your reply. Could the authors please post here the original uncropped scans for the images in Figure 5A, discussed in #2 above? ,