HBx-mediated decrease of AIM2 contributes to hepatocellular carcinoma metastasis
Authors: Shao-Hang Cai
Shi-Xun Lu
Li-Li Liu
Chun-Hua Wang
Hong Wang
Rong-Zhen Luo
Chris Zhiyi Zhang
Xia Yang
Jing-Ping Yun
Shi-Lu Chen
Dan Xie
Journal: Molecular Oncology (2017)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: EMT; absent in melanoma 2; hepatitis B virus; hepatocellular carcinoma.
Affiliations: Sun Yat-Sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China., Department of Pathology, Sun Yat-Sen University Cancer Center, Guangzhou, China.

Abstract:  Tumor metastasis is responsible for the high mortality rates in patients with hepatocellular carcinoma (HCC). Absent in melanoma 2 (AIM2) has been implicated in inflammation and carcinogenesis, although its role in HCC metastasis remains unknown. In the present study, we show that AIM2 protein expression was noticeably reduced in HCC cell lines and clinical samples. A reduction in AIM2 was closely associated with higher serum AFP levels, vascular invasion, poor tumor differentiation, an incomplete tumor capsule and unfavorable postsurgical survival odds. In vitro studies demonstrated that AIM2 expression was modulated by hepatitis B virus X protein (HBx) at transcriptional and post-translational levels. HBx overexpression markedly blocked the expression of AIM2 at mRNA and protein levels by enhancing the stability of Enhancer of zeste homolog 2 (EZH2). Furthermore, HBx interacted with AIM2, resulting in an increase of AIM2 degradation via ubiquitination induction. Functionally, knockdown of AIM2 enhanced cell migration, formation of cell pseudopodium, wound healing and tumor metastasis, whereas reintroduction of AIM2 attenuated these functions. The loss of AIM2 induced the activation of epithelial-mesenchymal transition (EMT). Fibronectin 1 (FN1) was found to be a downstream effector of AIM2, with its expression reversely modulated by AIM2. Silencing of FN1 significantly halted cell migration induced by AIM2 depletion. These data demonstrate that HBx-induced loss of AIM2 is associated with poor outcomes and facilitates HCC metastasis by triggering the EMT process. The results of the present study therefore suggest that AIM2 is a potential prognostic biomarker in hepatitis B virus-related HCC, as well as a possible therapeutic target for tumor metastasis. 

Retracted: No

PubPeer comments:
 And image in Figure 6 appears to have been duplicated, re-colored, cropped differently, and presented as something else (green boxes). ***  , 
 I am Shi-Lu Chen, the first author of the article titled u201cHBx-mediated decrease of AIM2 contributes to hepatocellular carcinoma metastasisu201d. Thank you for your valuable comments on our published paper. As we got the message, we immediately checked the original data of the experiments. After we checked the manuscript and the raw data, we found that some pictures indeed have been mismatched during the preparation of these panels. After taking the IHC and western blot data, there were lots of similar raw pictures which need further processing. During this process, we made a mistake when arranging the images. We sincerely apologized for these mistakes. We have corrected Figure 3E and Figure 6C in the attachment. On the positive note, the corrected results did not directly affect out hypothesis. We have contacted the editorial office of Molecular Oncology to issue a correction. Once more, we would like to express our most sincere gratitude for the comments to improve our manuscript. In the future, we will do our best, with a more rigorous attitude to review all current work and manuscript to prevent it from happening again. , 
 Thank you for responding. Unfortunately, the concerns about the veracity of the data in this paper seem to be more extensive than just these two figures. I have highlighted some additional potential problems and there may be more. Perhaps a retraction and resubmission would allow the authors to present better organized credible support for the conclusions. *** Supplemental Figure 4B (overlap and stretch)  *** Supplemental Figure 5D (overlap)  *** Figure 4A (band reuse)  , 
 #2 Any update on this? I also contacted the journal in February 2021. , 
 #4 **More seriously, the authors reused the same immunofluorescence results again and again, just in different colors to show the protein expressions in totally different experiments** **e.g. Figure 7C and Figure 8B**    , 
 **Also, more image reuses were found in these immunofluorescence experiments**    **Please be noted these IF experiments were in totally different series** **This type of image reuse is difficult to believe that it is caused by objective carelessness** ,