MiRNA 17 family regulates cisplatin-resistant and metastasis by targeting TGFbetaR2 in NSCLC
Authors: Youguang Pan
Zeyong Jiang
Siwen Li
Lu Dai
Wenfan Fu
Yijun Mo
Jian Zhao
Jun Yin
Haoda Huang
Journal: PLoS ONE (2014)
Discipline: Multidisciplinary
Keywords: MicroRNAs; cancer; chemotherapy resistance; metastasis; non-small-cell lung cancer
Affiliations: Department of Chest Surgery, Cancer Center of Guangzhou Medical University, Guangzhou, Guangdong, China.

Abstract:  MicroRNAs (miRNAs) have been proven to play crucial roles in cancer, including tumor chemotherapy resistance and metastasis of non-small-cell lung cancer (NSCLC). TGFβ signal pathway abnormality is widely found in cancer and correlates with tumor proliferation, apoptosis and metastasis. Here, miR-17, 20a, 20b were detected down-regulated in A549/DDP cells (cisplatin resistance) compared with A549 cells (cisplatin sensitive). Over-expression of miR-17, 20a, 20b can not only decrease cisplatin-resistant but also reduce migration by inhibiting epithelial-to-mesenchymal transition (EMT) in A549/DDP cells. These functions of miR-17, 20a, 20b may be caused at least in part via inhibition of TGFβ signal pathway, as miR-17, 20a, 20b are shown to directly target and repress TGF-beta receptor 2 (TGFβR2) which is an important component of TGFβ signal pathway. Consequently, our study suggests that miRNA 17 family (including miR-17, 20a, 20b) can act as TGFβR2 suppressor for reversing cisplatin-resistant and suppressing metastasis in NSCLC. 

Retracted: No

PubPeer comments:
 2019 expression of concern. https: journals.plos.org plosone article?id=10.1371 journal.pone.0222896 Following publication, concerns were raised about duplicate images in Figs 2, 3 and 4 of this article [1]. Specifically, in Fig 2A the top left two panels have a region of overlap and have been taken from the same microscopy image. In Fig 3C u201cA549 DDP by miRNAs mimicsu201d actin panel lanes 20a and 20b and in Fig 4A actin panel, the same image was used to represent the actin controls from two different experiments. The corresponding author has stated these were mistakes in the preparation of the original figures and provided revised figures. In Fig 2A, the u201cA549 DDP mimics-Conu201d panel was incorrect and has been replaced. Please see the correct Fig 2 and caption here. (A) Morphology of A549 and A549 DDP cells with or without miRNAs treatment. (B) mRNA and protein expression levels of E-cadherin and Vimentin in A549 and A549 DDP cells with or without miRNAs treatment were measured by RT-PCR and Western Blotting. (C) Transfer cells in migration assays were detected by transwell-chamber culture systems. Bar graphs show the number of migratory cells. *P 0.05. Results were representative of three experiments. https: doi.org 10.1371 journal.pone.0222896.g001 In Fig 3C, the u201cA549 DPP by mRNAs mimicsu201d actin panel was incorrect and has been replaced. Please see the correct Fig 3 and caption here. (A) Site-directed mutagenesis targeting potential miR-17, 20a, 20b binding sites (MUT) on the TGFu03b2R2 mRNA 3u2032-UTR-luciferase construct. (B) Luciferase activities significantly decreased in the WT TGFu03b2R2 mRNA 3u2032-UTR-luciferase plasmid transfected A549 cells after transfection of miR-17, 20a, 20b mimics. Effect was blocked in the MUT plasmid transfected A549 cells. (C) By RT-PCR and Western Blotting, the expression of TGFu03b2R2 increased significantly in A549 DDP cells compared with A549 cells. The expression of TGFu03b2R2 decreased in A549 DDP cells after transfection of miR-17, 20a, 20b mimics and increased in A549 cells after transfection of miR-17, 20a, 20b inhibitors. *P 0.05. Results were representative of three experiments. https: doi.org 10.1371 journal.pone.0222896.g002 In Fig 4A, the actin panel was incorrect and has been replaced. Please see the correct Fig 4 and caption here. (A) After inhibition of TGFu03b2R2 in A549 DDP cells by transfected with si-TGFu03b2R2, the EMT phenotype changed evidenced by over-expression of E-cadherin and down-regulation of Vimentin, and (B) the migratory capability also significantly decreased. Bar graphs show the number of migratory cells. (C) TGFu03b2R2 silenced induced by siRNA led to decrease cisplatin resistance, as shown by a significantly growth inhibition curve of cisplatin in A549 DDP cells. *P 0.05. Results were representative of three experiments. https: doi.org 10.1371 journal.pone.0222896.g003 The underlying blot images, microscopy images, and the data points underlying the charts in Figs 2, 3 and 4 are provided as Supporting Information, with the exception of the following items that are no longer available: The underlying blots for Fig 2B left hand panel E-cadherin, right hand panel u201cA549 by inhibitorsu201d Vimentin and actin, and middle panel u201cA549 DPP by mimicsu201d Vimentin. The underlying blot for Fig 2B middle panel u201cA549 DPP by mimicsu201d E-caderin. A blot from a replicate experiment carried out at the time of the original study is included in the Supporting Information. The datapoints underlying the chart in Fig 4A, which shows mRNA levels detected by RT-PCR. The authors have also provided the following clarifications and corrections: The samples used for actin and target protein detection were loaded into separate gels in the same order and marked on the corners of the gel and PVDF membrane, and for this reason, actin control bands appear in different blots to those of the target proteins. There is an error in the seventh sentence of the u201cmiR-17, 20a, 20b regulate EMT and migrationu201d subsection of the Results. The correct sentence is:Results showed that over-expression of miR-17, 20a, 20b could reduce migration in A549 DDP cells and inhibition of them could increase migration in A549 cells (Fig 2C). In the second sentence of the second paragraph of the Discussion section, references 6 and 12 are incorrectly placed and should not be included. The correct sentence is:Secondly, inhibition of miR-17, 20a, 20b increased cisplatin-resistant and migration of A549 cells, and over-expression of miR-17, 20a, 20b decreased cisplatin-resistant and migration of A549 DDP cells. The underlying data for Fig 1 are not available. This includes data from a microarray experiment performed by service provider LC Sciences. The Data Availability Policy in effect at the time of submission of the article required public deposition of microarray data, which is in accordance with standard practice in the field at the time. The PLOS ONE Editors issue this Expression of Concern to alert readers to the concerns about the unavailability of the microarray data, as well as the outlined concerns regarding the originally published images. , 
 Here are the concerns about Figure 2A (red boxes), and Figures 3 and 4 (green and cyan boxes), reported to the journal in April 2014: ![file] ,