miR-143 and miR-145 synergistically regulate ERBB3 to suppress cell proliferation and invasion in breast cancer
Authors: Ting Deng
Xi Chen
Xueliang Wang
Baorui Liu
Chihao Zhao
Xiujuan Gao
Ke Zen
Yi Ba
Rui Liu
Minghui Liu
Weixu Chen
Nan Wang
Yanqing Liu
Jialu Li
Hongwei Liang
Suyang Zhang
Xin Yan
Chen-Yu Zhang
Journal: Molecular Cancer (2014)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: ERBB3; miRNA; breast cancer; proliferation; invasion
Affiliations: Department of Gastrointestinal Medical Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy of Tianjin, Huanhuxi Road, Tiyuanbei, Tianjin 300060, China. .

Abstract:  Introduction: ERBB3, one of the four members of the ErbB family of receptor tyrosine kinases, plays an important role in breast cancer etiology and progression. In the present study, we aimed to identify novel miRNAs that can potentially target ERBB3 and their biological functions. Method: The expression levels of miR-143/145 and target mRNA were examined by relative quantification RT-PCR, and the expression levels of target protein were detected by Western blot. We used bioinformatic analyses to search for miRNAs that can potentially target ERBB3. Luciferase reporter plasmids were constructed to confirm direct targeting. Furthermore, the biological consequences of the targeting of ERBB3 by miR-143/145 were examined by cell proliferation and invasion assays in vitro and by the mouse xenograft tumor model in vivo. Results: We identified an inverse correlation between miR-143/145 levels and ERBB3 protein levels, but not between miR-143/145 levels and ERBB3 mRNA levels, in breast cancer tissue samples. We identified specific targeting sites for miR-143 and miR-145 (miR-143/145) in the 3'-untranslated region (3'-UTR) of the ERBB3 gene and regulate ERBB3 expression. We demonstrated that the repression of ERBB3 by miR-143/145 suppressed the proliferation and invasion of breast cancer cells, and that miR-143/145 showed an anti-tumor effect by negatively regulating ERBB3 in the xenograft mouse model. Interestingly, miR-143 and miR-145 showed a cooperative repression of ERBB3 expression and cell proliferation and invasion in breast cancer cells, such that the effects of the two miRNAs were greater than with either miR-143 or miR-145 alone. Conclusion: Taken together, our findings provide the first clues regarding the role of the miR-143/145 cluster as a tumor suppressor in breast cancer through the inhibition of ERBB3 translation. These results also support the idea that different miRNAs in a cluster can synergistically repress a given target mRNA. 

Retracted: No

PubPeer comments:
 Figure 3C appears to have some overlapping images, at different magnifications. Seems to be a recurring theme with this research group. *** ![file] , 
 - *[left]* Fig 1A from  Sirtuin 6 contributes to migration and invasion of osteosarcoma cells via the ERK1 2 MMP9 pathway ([Lin et al 2017] ). - *[right]* Fig 1A ![] , 
 #1 **Dear Actinopolyspora Biskrensis** Thank you very much for pointing out the error! I have carefully checked the raw data of the related figure. This error was made due to editing and assembly of the raw data. As shown in the following clarification, I have checked the raw data and figured out what has happened. I apologize to our readers for any confusion this unintentional error might have caused. I have contacted the editorial office of Molecular Cancer for correction and the corrected data will be published soon. Because I don t have the institutional email address to register in the Pubpeer, I can only reply to your question as a comment. Sincerely, Xin Yan **The contacted email with the editorial office of Molecular Cancer** ![file] **The correct figure** ![file] Figure 3 miR-143 145 regulate proliferation and invasion of breast cancer cells by targeting ERBB3. (A and B) MTT viability assayswere performed 12, 24, 36, 48 and 60 h after the transfection of MCF-7 (A) and MBA-MD-231 (B) cells with pre-miR-control, pre-miR-143, pre-miR-145 or pre-miR-143 145 (left panel), or with anti-miR-control, anti-miR-143, anti-miR-145 or anti-miR-143 145 (middle panel), or witheither pre-miR-control, pre-miR-143 145 or both pre-miR-143 145 and the ERBB3 overexpression vector (right panel). (C-F) Transwell invasionassays were performed after the transfection of MCF-7 (C and D) and MBA-MD-231 (E and F) cells with pre-miR-control, pre-miR-143, pre-miR-145 or pre-miR-143 145 (upper panel), or with anti-miR-control, anti-miR-143, anti-miR-145 or anti-miR-143 145 (middle panel), or with either pre-miR-control,pre-miR-143 145 or both pre-miR-143 145 and the ERBB3 overexpression vector (lower panel). C and E: representative image; D and F: quantitativeanalysis. * P 0.05; ** P 0.01. **The detailed clarification** The relevant data of this article is stored in the folder named u201cERBB3u201d. ![file] The original data of this article is stored in a folder named u201cu539fu59cbu6570u636eu201duff0cwhich means raw data. ![file] The original data of transwell invasion assays in this article is stored in the folder named  transwell . ![file] Three to eight images were collected from the chamber of each group.u201cPVu201dmeans to pre-miRNA-143 145+ERBB3 vector. u201cA1u201d means to anti-miRNA-143. u201cA2u201d means to anti-miRNA-145. u201cA3u201d means to anti-miRNA-143 145. u201cCu201dmeans to control. u201cP1u201dmeans to pri-miRNA-143. u201cP2u201dmeans to pri-miRNA-145. u201cP3u201dmeans to pri-miRNA-143 145. In the preparation of the article, due to our mistakes, the pictures that belong to pre-miR-143 and anti-miR-143 (marked as P1-1 and A1-4) are wrongly marked as P3-3 and 1-Pv-1. ,