SMG-1 inhibition by miR-192/-215 causes epithelial-mesenchymal transition in gastric carcinogenesis via activation of Wnt signaling
Authors: Mengting Yang
Song Li
Xinmin Fan
Zhendan He
Ruibin Yan
Zhe Jin
Xianling Feng
Kuan Li
Liang Wang
Stephen J Meltzer
Shanni Li
Yanqiu Zhao
Yulan Cheng
Xiaojing Zhang
Si Chen
Huijuan Lin
Yanjie Wei
Xi Liu
Shiqi Deng
Huimin Yu
Yin Peng
Zhenfu Zhao
Yong Huang
Journal: Cancer Medicine (2018)
Discipline: CLINICAL MEDICINE
Keywords: EMT; SMG-1; Wnt signaling pathway; gastric cancer; miR-192/-215.
Affiliations: Department of Pathology, The Shenzhen University School of Medicine, Shenzhen, Guangdong, 518060, China., Shenzhen Key Laboratory of Micromolecule Innovatal Drugs, Shenzhen Key Laboratory of translational Medicine of Tumor, Guangdong Key Laboratory for Genome Stability & Disease Prevention, Shenzhen University School of Medicine, Shenzhen, Guangdong, China., Guangdong Province Key Laboratory of Molecular Oncologic Pathology, Guangzhou, Guangdong, China., Department of Pathology, Wuhan University School of Basic Medical Sciences, Hubei, China., Laboratory of Chemical Genomics, The Shenzhen Graduate School of Peking University, Shenzhen, Guangdong, China., Department of Medicine/GI Division, Johns Hopkins University, Sidney Kimmel Comprehensive Cancer Center, Baltimore, Maryland., Department of Pathology and Pathophysiology, The Guangzhou Medical University, Guangzhou, Guangdong, China., Center for High Performance Computing, Shenzhen Institutes of Advanced Technology, Shenzhen, Guangdong, China.

Abstract:  SMG-1,a member of the phosphoinositide kinase-like kinase family, functioned as a tumor suppressor gene. However, the role of SMG-1 in GC remain uncharacterized. In this study, regulation of SMG-1 by miR-192 and-215, along with the biological effects of this modulation, were studied in GC. We used gene microarrays to screening and luciferase reporter assays were to verify the potential targets of miR-192 and-215. Tissue microarrays analyses were applied to measure the levels of SMG-1 in GC tissues. Western blot assays were used to assess the signaling pathway of SMG-1 regulated by miR-192 and-215 in GC. SMG-1 was significantly downregulated in GC tissues.The proliferative and invasive properties of GC cells were decreased by inhibition of miR-192 and-215, whereas an SMG-1siRNA rescued the inhibitory effects. Finally, SMG-1 inhibition by miR-192 and-215 primed Wnt signaling and induced EMT. Wnt signaling pathway proteins were decreased markedly by inhibitors of miR-192 and-215, while SMG-1 siRNA reversed the inhibition apparently. Meanwhile, miR-192 and-215 inhitibtors increased E-cadherin expression and decreased N-cadherin and cotransfection of SMG-1 siRNA reversed these effects. In summary, these findings illustrate that SMG-1 is suppressed by miR-192 and-215 and functions as a tumor suppressor in GC by inactivating Wnt signaling and suppressing EMT. 

Retracted: No

PubPeer comments:
 The DAPI Edu images in this paper look unexpected in that they do not match each other. DAPI binds to DNA, while Edu is incorporated into DNA of dividing cells, so both stains should at least partially co-locate. But the merged images in Figures 2 and 6 do not appear to match each other at all. Here are Figure 2F and 2G to show the non-colocational merged images: ![file] ![file] Also,  Author order was determined by drawing straws made me smile. , 
 Question 1: The DAPI Edu images in this paper look unexpected in that they do not match each other. DAPI binds to DNA, while Edu is incorporated into DNA of dividing cells, so both stains should at least partially co-locate. But the merged images in Figures 2 and 6 do not appear to match each other at all. Responseuff1aThank you for your interest in our work and your contribution to protecting the accuracy of scientific records and scientific integrity; they have always been the most important to me. After receiving your inquiry notice, I have listed it as our top priority, and immediately take measures to investigate your questions to the team and partners, and carefully recheck our manuscripts, original data and laboratory records. Indeed, as you said, DAPI binds to DNA, while EdU is incorporated into DNA of dividing cells, so both stains should at least partially co-locate. And I highly recognize your professional evaluation. In addition, if the blue DAPI and red EdU are co-located, a new color, purplish red, will be displayed in the merged image, which is obviously different from red and blue. After carefully checking Figures 2 and 6 in our article, we found that there was no problem with the original figures uploaded to the Journal of Virology after the last modification, but this part of the merge image information was lost in the final published PDF file, which may be due to the loss of image quality in the process of converting Figures into PDF, which is beyond our control. In order to dispel your doubts, we attach the original pictures submitted to the Journal of Virology, please check the attachment. With regard to the complete mismatch of the pictures you proposed, we think the purple red in the merged image can explain your doubts. There is co-localization of DAPI and EdU in our merged image. Moreover, the exposure of DAPI and EdU is different, because the fluorescence intensity of EdU is often several times that of DAPI, so the exposure of DAPI and EdU is different. If the exposure is the same, the exposure intensity of either of them will be too strong or too weak. Therefore, we can only ensure that all EdU exposures are consistent and all DAPI exposures are consistent. So, the co-location form of the DAPI and EdU can not be completely consistent, and there will be slight differences, I hope to get your understand. Finally, thank you again for your professional evaluation and rigorous attitude, which every scientific research worker should deeply learn for you. Question 2: Also,  Author order was determined by drawing straws made me smile. Response: The issue of  author order was determined by drawing straws mentioned by you is not decided by us, but according to the provisions of the journal. In fact, the contributions made by the first author and the second author in our article are still different, but we are also very helpless because of the requirements of the journal, which not only makes you smile, but also makes us smile. You can look at other articles in Journal of Virology, and these articles basically have this sentence. We have also made a screenshot of this requirement of the journal as an attachment, please check it. This is clearly stipulated in the journal requirements, so I hope to get your understand. Fig. 2F and 2G ![file] Fig.6G ![file] JV-authorship ![file] ,