MicroRNA-330-3p promotes cell invasion and metastasis in non-small cell lung cancer through GRIA3 by activating MAPK/ERK signaling pathway
Authors: Rui-Guang Zhang
Ji-Hua Dong
Qian Cai
Rui Zhou
Yu Hu
Xi-Can Gao
Xiao-Rong Dong
Fan Tong
Gang Wu
Chun-Hua Wei
Journal: Journal of Hematology & Oncology (2017)
Discipline: CLINICAL MEDICINE
Keywords: DNA methyltransferase; GRIA3; Invasion and metastasis; MAPK/ERK signaling; NSCLC
Affiliations: Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 JieFang Avenue, Wuhan, 430022, People's Republic of China., Experimental Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277, JieFang Avenue, Wuhan, 430022, People's Republic of China., Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 JieFang Avenue, Wuhan, 430022, People's Republic of China., Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 JieFang Avenue, Wuhan, 430022, People's Republic of China. .

Abstract:  Background: Brain metastasis (BM) is associated with poor prognosis in patients with non-small cell lung cancer (NSCLC). Recent studies demonstrated that microRNA-330-3p (miR-330-3p) was involved in NSCLC brain metastasis (BM). However, the exact parts played by miR-330-3p in BM of NSCLC remain unknown. Discovery and development of biomarkers and elucidation of the mechanism underlying BM in NSCLC is critical for effective prophylactic interventions. Here, we evaluated the expression and biological effects of miR-330-3p in NSCLC cells and explored the underlying mechanism of miR-330-3p in promoting cell migration and invasion in NSCLC. Methods: Stable over-expression and knockdown of miR-330-3p in NSCLC cells was constructed with lentivirus. Expression levels of miR-330-3p in NSCLC cells were quantified by quantitive real-time PCR (qRT-PCR). The effects of miR-330-3p on NSCLC cells were investigated using assays of cell viability, migration, invasion, cell cycle, apoptosis, western blotting, immunohistochemical, and immunofluorescence staining. A xenograft nude mouse model and in situ brain metastasis model were used to observe tumor growth and brain metastasis. The potential target of miR-330-3p in NSCLC cells was explored using the luciferase reporter assay, qRT-PCR, and western blotting. The miR-330-3p targets were identified using bioinformatics analysis and verified by luciferase reporter assay. The correlation between GRIA3 and DNA methyltransferase (DNMT) 1 and DNMT3A was tested by RT-PCR, western blotting, and co-immunoprecipitation (IP). Results: miR-330-3p was significantly up-regulated in NSCLC cell lines. MTT assay, transwell migration, and invasion assays showed that miR-330-3p promoted the growth, migration, and invasion of NSCLC cells in vitro and induced tumor growth and metastasis in vivo. Luciferase reporter assays showed that GRIA3 was a target of miR-330-3p. qRT-PCR and western blotting exhibited that miR-330-3p promoted the growth, invasion, and migration of NSCLC cells by activating mitogen-activated protein kinase (MAPK)/extracellular-regulated protein kinases (ERK) signaling pathway. Furthermore, miR-330-3p up-regulated the total DNA methylation in NSCLC cells, and co-IP-demonstrated GRIA3 was directly related with DNMT1 and DNMT3A. Conclusions: miR-330-3p promoted the progression of NSCLC and might be a potential target for the further research of NSCLC brain metastasis. 

Retracted: No

PubPeer comments:
 Figure 2f appears to have a repeated image. This seems to be a recurring issue for this [set of authors] . *** ![file] , 
 https: retractionwatch.com 2013 11 27 cancer-researchers-retract-two-papers-in-the-jbc https: retractionwatch.com 2016 02 08 2014-ori-finding-results-in-retraction-of-cancer-paper-with-manipulated-images https: retractionwatch.com 2014 12 14 ori-sanctions-former-university-chicago-ucsf-scientists-faking-findings , 
 #1 First of all, we examined the indicated Figure 2f and agree with the assessment that repeated images were used. We are grateful for that you alert us of such errors present in Figure 2. To look into the nature and cause of such errors, we immediately checked the lab experiment record books that contain the lab notes and the electronic original data related to this part of the study. We found the original lab records in the lab note book that described this part of the study (see the inserted photocopy of the lab record). Since the lab notes were written in Chinese, we made a line-by-line translation of the lab note for your comprehension and review. Translated version: Line1: after adding Brefeldin A Line2: the secreted S100A8 from OL-N.C. and POL-Sec23a-OE were significantly decreased Line3-4: conclusion: when Sec23a was highly expressed, the secreted S100A8 was increased, when the transportation between ER and Golgi apparatus was cut off with Brefeldin A, the secreted S100A8 was significantly decreased Line5: Date: October 26th, 2018 Line6: Content: to detect the relationship between autophagy and Sec23a with TEM Line7: Location: room 310, Life Sciences Institute Line8: Staff: Zhiwei Sun Line16: TEM is an important standard for detecting autophagy Line17: TEM can see the bilayer membrane structure of autophagosomes Line18: The contents in autophagosomes vary according to different stages of autophagy Line19: early stage: complete light-colored organelles (mainly mitochondria) Line20: late stage: dark-colored protein degradation products ![file] Our observations and responses based on the examination of the original lab records: The printed figure summarizes TEM analysis of the autophagesome in the lab records is identical to that in Figure 2f in the published paper. Thus, the error occurred and documented in the lab book. We examined the original data that contain all TEM images taken and found other images for all the experimental groups that can be used for the publication. Since the figure appeared as Figure 2f was made after the TEM analysis was completed, before the write up of the manuscript and was recorded in the lab book, thus the use of repeated images was most likely a careless and unintentional mistake. Nevertheless, all co-authors failed to identify the error during reviewing of the manuscript. The student who conducted this part of the study and made the Figure failed to verify the figure made with the original data which is a required in our laboratory practice before submitting the manuscript for peer review. When request for making the corrections, we will provide the Journal with the original data of all images taken to show that the experiments had been carefully done with multiple repeats. We revised Figure 2f based on the original data for your examination. The revised figure: ![file] , 
 Published: 10 March 2022 The original version of this article unfortunately contained a mistake. After publication of the article, the authors noticed an unintentional error in Fig. 2f. As shown in Fig. 2f, the representative transmission electron microscopy image of u201cOL-shSec23au201d was identical to the image of u201cPOL-vectoru201d. The authors have confirmed that the image of u201cOL-shSec23au201d was mistakenly presented in the original Fig. 2f. This error was caused by unintentionally misplacing the representative image. Nevertheless, this error does not change the conclusion or the discussion of the article. The authors apologize for any inconvenience this error may cause. The corrected Fig. 2f is provided below. The original article has been corrected. ![file] ,