Fusobacterium nucleatum Promotes Metastasis in Colorectal Cancer by Activating Autophagy Signaling via the Upregulation of CARD3 Expression
Authors: Yan Chen
Pan Cao
Yongyu Chen
Yunchao Deng
Yun Huang
Xiangsheng Fu
Na Zhan
Wenhao Su
Weiguo Dong
Jixiang Zhang
Journal: Theranostics (2020)
Discipline: CLINICAL MEDICINE
Keywords: colorectal cancer; gene regulation; gene targeting; microbe.
Affiliations: Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China., Key Laboratory of Hubei Province for Digestive System Disease, Wuhan, Hubei Province, China., Central Laboratory, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China., Department of Gastroenterology, The Affiliated Hospitalof North Sichuan Medical College, Road Wenhua 63#, Region Shunqing, Nanchong City 637000, China., Center for Epigenetics & Disease Prevention, Institute of Biosciences and Technology, Texas A&M University, Houston, TX77030, USA.

Abstract:  Aims: We aimed to measure the abundance of Fusobacterium nucleatum (F. nucleatum) in colorectal cancer (CRC) tissues from patients and to uncover the function of this bacterium in colorectal tumor metastasis. Methods: We collected metastatic and non-metastatic CRC tissues to analyze F. nucleatum abundance. Cells were incubated with F. nucleatum or chloroquine (CQ) or were transfected with CARD3-targeting siRNA; the expression of mRNAs and proteins was then measured. CRC cells stably transfected with shRNA-luc were mixed with F. nucleatum and intravenously injected into BALB/cJ mice. APCMin/+, CARD3-/-and CARD3wt C57BL mice were given F. nucleatum; some mice were given azoxymethane (AOM) and dextran sodium sulfate (DSS). Results: F. nucleatum was abundant in CRC tissues from patients with metastasis. F. nucleatum infection increased CRC cell motility and upregulated the expression of CARD3, LC3-II, Beclin1 and Vimentin, and downregulated the expression of E-cadherin and P62 in CRC cells. These effects were attenuated by treatment with CQ, siCARD3 or both. APCMin/+ mice gavaged with F. nucleatum developed more aggressive tumors than control mice. After AOM/DSS administration, the colorectums of CARD3-/- mice had fewer tumors than those of control mice. Tumors from CARD3-/- mice had lower levels of LC3-II and Beclin1 and higher levels of P62 than those from control mice. BALB/cJ mice injected with both CT26-luc cells and F. nucleatum formed more metastases than control mice. CQ treatment, CARD3 knockdown or both reduced the ability of CT26-luc cells to form metastases in vivo. Conclusions: F. nucleatum is enriched in CRC tissues from patients with metastasis. F. nucleatum orchestrates CARD3 and autophagy to control CRC metastasis. Measuring and targeting F. nucleatum and its associated pathways will yield approaches for the prevention and treatment of CRC metastasis. 

Retracted: No

PubPeer comments:
 Figure 3B and Figure 6A appear to have images which overlap. *** ![file] ![file] , 
 Figure 3B appears to have two images which overlap. *** ![file] , 
 Figure 3B and Figure 6A appear to have a repeated image and another two images which overlap. *** ![file] ![file] *** ![file] ![file] , 
 6B overlap. Iu2019ll stop there. *** ![file] , 
 Thanku00a0youu00a0foru00a0the commentsu00a0concerningu00a0ouru00a0manuscript entitledu00a0u201cFusobacterium nucleatum Promotes Metastasis in Colorectal Cancer by Activating Autophagy Signaling via the Upregulation of CARD3 Expressionu201d. The carelessness in combining the images caused the repetition of the images. It is indeed because there are more research data, and these problems have been caused by negligence. We are very sorry that our negligence caused the problem. We have replaced themu00a0with the correct imagesu00a0and will contact with the  Theranostics to correct it. We feel fortunate that the commentsu00a0will help improve our manuscript and we are once again deeply sorry for the problem caused by our negligence. , 
 Consolidated illustration of 3A, 6A, and 6B. https: imgur.com gallery dWHcmyI , 
 We thank the kind reader for indicating the mistake regarding to the representative pictures of trans well experiments in our paper. We did make a mistake of mixing up the transwell pictures of HCT116 in Figure3B and SW480 inFigure 6B. We have corrected the wrong pictures and provided the corrected Figure 3B and Figure 6B as shown as follow. We also applied to the editor of  Theranostics to help us correct the misplaced pictures. We apologize for the negligence in our work! We thank the reader again and thank the staff working for PuPeer very much! ![file] ![file] , 
 #7 Yes, well... Even your corrected figure appears to include an overlap (below). I suggest the senior author request a retraction and attempt to submit a new paper with more careful results. Given the propensity of what appears to be the same research group carelessly commingling images (see thread below), I would think it is difficult to trust the research results from this lab. I will be interested in seeing how the journal digests these facts. u201cNegligenceu201d seems insufficient to describe problems of this sort. I do apologize for corresponding in English, as I assume that is not the authorsu2019 native language. https: pubpeer.com search?q=authors%3A%22Weiguo+Dong%22 ![file] , 
 Erratum Published: 2022.01.01 The authors regret that the original version of our paper [1] unfortunately contained two inappropriate representative images of WB. Noting that WB images (the Vimentin of ATCC10953 and the CARD3 of F01) in the first submitted version were correct. For presenting high resolution images according to one of the reviewer s suggestion, we assembled inappropriate images in our revised manuscript (Figure 1I and Figure 4H). We regret that at the time of figure assembly, we choose representative images by mistake. Besides, we apologize for the original version of our paper unfortunately contained some inappropriate representative images of transwell. We mixed up some of them, which caused some confusion in Figure 3B, Figure 6AB and Supplementary Figure 1B. Although we have checked the paper before submission, we have not been careful enough to find out the problems in time. We apologize for our careless at the time of figure assembly. We confirm that it would not affect any results and conclusions of the paper. The authors apologize for any inconvenience that this error may have caused. ![file] ![file] ![file] ,