<p>microRNA-612 suppresses the malignant development of non-small-cell lung cancer by directly targeting bromodomain-containing protein 4</p>
Authors: Fanwu Kong
Chengcheng Yang
Xiaowen Kang
Qiushuang Liu
Wei Zhang
Shijie Wu
Xiaomei Wu
Journal: OncoTargets and Therapy (2019)
Discipline: CLINICAL MEDICINE
Keywords: biomarker indicator; bromodomain-containing protein 4; microRNA-612; non-small-cell lung cancer.
Affiliations: Department of Respiration, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, People's Republic of China., Department of Nephrology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, People's Republic of China., Department of Respiration, General Hospital of Daqing Oil Field, Daqing, Heilongjiang 163000, People's Republic of China., Department of Pharmacy, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, People's Republic of China., Department of Respiration, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150000, People's Republic of China.

Abstract:  Background: Aberrant expression of microRNAs (miRNAs) in non-small-cell lung cancer (NSCLC) has been reported. Dysregulation of miRNAs exerts tumor-suppressing or tumor-promoting actions on the pathology and biological behaviors of NSCLC. miR-612 is associated with many types of human cancer; however, the expression, potential roles, and regulatory mechanisms of miR-612 in NSCLC remain unclear. Material and methods: Here, the expression level of miR-612 in NSCLC tissue specimens and a panel of cell lines were evaluated by RT-qPCR. Cell-Counting Kit 8, flow cytometry, Transwell migration and invasion, and in vivo tumor growth assays were performed to determine the functional role of miR-612 in malignant phenotypes of NSCLC cells. The molecular mechanism underlying the tumor-suppressive roles of miR-612 in NSCLC was investigated. Results: miR-612 was expressed at low levels in NSCLC, and low miR-612 expression was significantly correlated with TNM stage and lymph node metastasis. NSCLC patients with low miR-612 expression had shorter overall survival rate than those with high levels. Exogenous miR-612 expression decreased proliferation, migration, and invasion, and promoted apoptosis of NSCLC cells in vitro. miR-612 upregulation hindered NSCLC tumor growth in vivo. Bromodomain-containing protein 4 (BRD4) was confirmed as a direct target gene of miR-612 in NSCLC cells. BRD4 was obviously overexpressed in human NSCLC tissues and inverse correlated with miR-612 expression. Inhibition of BRD4 expression simulated the tumor-suppressive functions of miR-612 overexpression in NSCLC cells. Reintroduction of miR-612 expression abrogated the miR-612-mediated suppressive effects on NSCLC cells. BRD4 upregulation inhibited activation of the PI3K/Akt pathway in NSCLC cells in vitro and in vivo. Conclusion: This study supports the first evidence that miR-612 exerts tumor-suppressive roles in the aggressive behaviors of NSCLC cells in vitro and in vivo through direct targeting BRD4 and deactivating the PI3K/Akt pathway. Thus, miR-612 might be a promising target for anticancer therapies in patients with NSCLC. 

Retracted: Yes

PubPeer comments:
 Figs 2C, 5C, 6C.  Apoptotic cells were identified using flow cytometry analysis, and the results showed that miR-612 overexpression increased the apoptosis of H522 and A549 cells ;  The ... apoptosis in indicated cells were examined using ... flow cytometry analysis ;  apoptosis ... of H522 and A549 cells treated as described were determined using ... flow cytometry analysis . ![file] Purple box marks a panel that also appears in Figs 2D, 4D, 5D of [Zhang et al (2019)] . ![file] ![file] I did not search further for other matches, or matches with other papers from this oeuvre. https: docs.google.com spreadsheets d 1X401JM6CcpTSkbKsPsOl20pRKQ-ns1S_J5pTwtBjqPw edit?usp=sharing , 
 *[right]* Fig 8A.  Representative images of tumor xenografts derived from nude mice implanted with agomiR-612 or agomiR-NCtransfected H522 cells . *[left]* Fig 8A from [Song et al (2019)] .  Images of tumor xenografts derived from miR-564 mimics or miR-NC-transfected TPC-1 cells . Rotated through 90 deg. *[middle]* Fig 8A from [Yun et al (2019)] .  Photographs of tumor xenografts derived from Hep3B cells transfected with miR-548b mimics or negative control (NC) mimics . ![file] Coloured boxes indicate xenograft tumors that appear to be the same (slightly rotated). , 
 Fig 5C in isolation. ![] Coloured boxes mark matching panels in Figs 2D, 3D of [Yu et al (2020)] . ![] , 
 Dear Hoya Camphorifolia Thanks for your comments and noticing our mistake. We apologize for the delay in responding to you. We are also very sorry for these problems Our team are checking the original results and pictures. To verify our research results, we also will repeat some experiments and contact the journal for erratum. We do appreciate your careful review, and will give further response as soon as possible. Xiaowen kang , 
 Dear Hoya Camphorifolia Thanks for your comments and noticing our mistake. We apologize for the delay in responding to you. We are also very sorry for these problems Our team are checking the original results and pictures. To verify our research results, we also will repeat some experiments and contact the journal for erratum. We do appreciate your careful review, and will give further response as soon as possible. Xiaowen kang , 
 ![file] Left-hand panel is from Fig 5C. Central panel from Fig 3D of [Yu et al (2020)] . Right-hand panel from Fig 6B of [Yin et al (2020)] . Here in context: ![file] , 
 Retracted, [3 September 2021] . --------------------------------------------- The Editor and Publisher of *OncoTargets and Therapy* wish to retract the published article. Concerns were raised regarded the alleged duplication of images in Figures 5C and 8A with similar images from unrelated articles, specifically: - Figure 5C, panel A549 BRD4 siRNA appears to have been duplicated with a similar image in Figure 2D, panel H522 miR-802 mimics from Zhang et al, 2019 . - Figure 5C panels H522 and A549; NC siRNA, BRD4 siRNA appear to have been duplicated with similar images in Figures 2D, panel pcDNA3.1; pHAND2-AS1 and 3D panels si-NC; siHAND2-AS1 from Yu et al, 2020 . - Figure 8A, agomir-NC appears to have been duplicated with a similar image in Figure 8A, NC mimics from Yun et al, 2019 . The authors responded to our queries but were unable to provide an explanation for the alleged image duplication and could not provide the original raw data for their study and have therefore agreed to retract the article. Our decision-making was informed by our policy on publishing ethics and integrity and the COPE guidelines on retraction. The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as u201cRetractedu201d. ,