Effects of ebv‚ÄêmiR‚ÄêBART7 on tumorigenicity, metastasis, and TRAIL sensitivity of non‚Äêsmall cell lung cancer
Authors: Fei Liao
Lei Wang
Li Zhou
Li-Fa Liu
Ju Wang
Journal: Journal of Cellular Biochemistry (2019)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: TNF-related apoptosis-inducing ligand; apoptosis; ebv-miR-BART7; invasion; metastasis
Affiliations: Department of Thoracic Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China., Department of Thoracic Surgery, The Affiliated Hospital of Shandong Medical College, Linyi, Shandong, China., The Central Operating Room, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China.

Abstract:  Objective: To investigate how the Epstein-Barr virus (EBV) encoded microRNA BART7 (miR-BART7) affects tumorigenicity, metastasis, and TRAIL sensitivity of non-small cell lung cancer (NSCLC). Methods: Real time-polymerase chain reaction was performed to detect miR-BART7 expression in NSCLC cell lines. A549 and Calu-1 cells transfected with miR-BART7 inhibitors/mimics were used to do the in-vitro experiments, including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Annexin V-fluorescein isothiocyanate/PI, wound-healing, transwell, clonogenic assays, Western blot analysis, and anchorage-independent growth assay. Additionally, mice were used to inject A549 cells infected with miR-BART7 inhibitors to observe the tumorigenicity and metastasis of NSCLC. Results: TRAIL-resistant NSCLC cell lines (H460R, A549, Calu-1, and H1299) exhibited higher miR-BART7 rather than sensitive H460 and H292 cells. After transfected with miR-BART7 inhibitors, we observed an inhibition in proliferation, migration, invasion, and colony formation, but an enhancement in apoptosis as well as expressions of caspase-3 and caspase-8 in A549 and Calu-1 cells. Besides, TRAIL elevated the migration, invasion, and anchorage-independent growth of A549 cells, which was reversed by silencing DR4 and DR5 (siDRs). However, miR-BART7 inhibitors could reduce migration, invasion, and transformation potential of TRAIL treated A549 cells. Moreover, the expression of transforming growth factor-beta 1 (TGFŒ≤1) could be decreased by miR-BART7 inhibitors with or without TRAIL treatment. Moreover, the tumor growth, epithelial-to-mesenchymal transition, and metastasis was suppressed and tumor-free survival was extended after injection of A549-miR-BART7 inhibitors. Conclusion: Inhibition of miR-BART7 exerted inhibitory effects on cell proliferation, migration, invasion, and colony formation, consequently facilitating cell apoptosis and raising TRAIL sensitivity, providing a new therapeutic target in NSCLC. 

Retracted: Yes

PubPeer comments:
 Fig 3A.  Effects of miRu2010BART7 on apoptosis of TRAIL resistant NSCLC cells. Notes: Au2010D, Effects of miRu2010BART7 on apoptosis of A549 cells (A,B) ... were detected using Annexin Vu2010FITC PI staining . NC and Inhibitor panels are more similar than one might expect.  The Inhibitor panel includes more points in the upper right quadrant, including a constellation of cells that appears to have been sourced from the upper right quadrant of the Blank panel (with a vertical flip). Green box around the Blank panel is to highlight the surprising similarity to the Blank panel of fig 5B of [Qu et al (2016)] :  , 
 Fig 3C.  Caluu20101 cells .  At right, Mimics panel appears similar to 5B(c) of [Qu et al (2016)] , and almost identical to the NC panel of Fig 7A, [Wang et al (2016)] .  Orange and yellow boxes call attention to the similarity of the upper left quadrants of two panels to panels in 7A of Wang et al,  and panels of Fig 6 of [Ao et al (2017)] .  , 
 Fig 7A,C.  Effects of miRu2010BART7 on tumorigenicity of NSCLC cells. Notes: Au2010D, Clonogenic assay was conducted to detect colony formation after transfection in A549 (A,B) and Caluu20101 cells (C,D).  For comparison, Fig. 4 from [Li et al (2016)] .  Effect of miR-196a inhibitor transfection on proliferation of A549 DDP cells (au2013c cell clone formation in blank group, miR-NC group and anti-miR-196a .  Other colors correspond to panels of *[Above]* Fig 5C from [Liu et al (2017)] .  Effect of LncRNA-HOST2 expression on the proliferation of SMMC-7721 cells evaluated by clone-forming assay ... (C) Clone number *[Below]* Fig 5 from [Wang et al (2017)] .  Cell growth (A) and colony number (B) of MG-63 cells as detected using the colony assay .  UPDATE: Another sighting! Fig 4B of [Ba et al (2017)] .  colony formation ability of HCT-116 cells after 21 d of culture was examined by soft agar colony formation assay, which was performed 48 h after transfection .  , 
 #2 Another sighting! Fig 6A of [Zhu et al (2018)] .  , 
 Fig 3A,C again:   Indistringuishable from Fig 4 of [Li et al (2015)] .  Flow cytometry analysis showing the effect of miR-150 on stimulating MG63 and SOSP-9607 cell apoptosis. The apoptosis rates of MG63 and SOSP-9607 cells were significantly higher in the miR-150 group than in the blank group, but apoptosis rates of MG63 and SOSP-9607 cells were markedly lower in the inhibitor group than in the blank group.  , 
 Fig 4C.  Effects of miRu2010BART7 on the ... invasion of TRAILu2010resistant NSCLC cells. Notes: The migration and invasion of ... C,D, Caluu20101 cells were detected by using ... Transwell invasion assays .  Orange, brown, magenta rectangles indicate overlaps with Fig 7A of [Xiong et al (2018)] .  Ishikawa and RL95-2 cell invasion and migration abilities in blank, NC, miR-183 mimic, miR-183 inhibitor, CPEB1, and miR183 mimics + CPEB1 groups for Ishikawa and RL95-2 cells, detected by the transwell assay .  Other manifestations are linked in that other thread. , 
 Retracted [19 July 2021] . ---------------------------------------------- The above article, published online on 19 December 2018 in Wiley Online Library has been retracted by agreement between the journal s Editor in Chief, Prof. Dr. Christian Behl, and Wiley Periodicals LLC. The retraction has been agreed following an investigation based on allegations raised by a third party. A detailed investigation revealed that several image elements of the experimental data were published elsewhere in a different scientific context. Thus, the editors consider the conclusions of this article to be invalid. ,