Sec23a mediates miR-200c augmented oligometastatic to polymetastatic progression
Authors: H Rosie Xing
Ting Ye
Doudou Liu
Shixia Zhou
Jian Zhou
Zhiwei Sun
Jianyu Wang
Junling Tang
Jingyuan Li
Journal: EBioMedicine (2018)
Discipline: CLINICAL MEDICINE
Keywords: Oligometastasis; Polymetastasis; Sec23a; Secrotome; miR-200c.
Affiliations: Laboratory of Translational Cancer Stem Cell Research, Institute of Life Sciences, Chongqing Medical University, Chongqing, China., Institute of Neuroscience, Chongqing Medical University, Chongqing, China. Electronic address: ., Laboratory of Translational Cancer Stem Cell Research, Institute of Life Sciences, Chongqing Medical University, Chongqing, China. Electronic address: ., Laboratory of Translational Cancer Stem Cell Research, Institute of Life Sciences, Chongqing Medical University, Chongqing, China; School of Biomedical Engineering, State Key Laboratory of Ultrasound Engineering in Medicine, Chongqing Medical University, Chongqing, China. Electronic address: .

Abstract:  Background: Cancer treatment is based on tumor staging. Curative intent is only applied to localized tumors. Recent studies show that oligometastatic patients who have limited number of metastases may benefit from metastasis-directed local treatments to achieve long-term survival. However, mechanisms underlying oligometastatic to polymetastatic progression remains elusive. Methods: The effects of miR-200c and Sec23a on tumor metastasis were verified both in vitro and in vivo. The secretome changes were detected by mass spectrometry. Findings: We established a pair of homologous lung-metastasis derived oligometastatic and polymetastatic cell lines from human melanoma cancer cell line M14. Using the two cell lines, we have identified Sec23a, a gene target of miR-200c, suppresses miR-200c augmented oligometastatic to polymetastatic progression via its secretome. Firstly, miR-200c over-expression and Sec23a interference accelerated oligometastatic to polymetatic progression. Secondly, Sec23a functions downstream of miR-200c. Thirdly, mass spectrometric analysis of the secretory protein profile suggests that Sec23a-dependent secretome may impact metastatic colonization by modifying tumor microenvironment. Fourthly, the survival analysis using The Cancer Genome Atlas database shows Sec23a as a favorable prognostic marker for skin cutaneous melanoma, supporting the clinical relevance of our findings. Interpretation: The finding that Sec23a is a suppressor of oligometastatic to polymetastatic progression has clinical implications. First, it provides a new theoretical framework for the development of treatments that prevent oligometastasis to polymetastasis. Second, Sec23a may be used as a favorable prognostic marker for the selection of patients with stable oligometastatic disease for oligometastasis-based local therapies of curative intent. FUND: National Natural Science Foundations of China. 

Retracted: No

PubPeer comments:
 Figure 1 appears to have one set of images that overlap and one set of repeated images. ***  , 
 Figure 4c appears to have two images which overlap. This seems to be a recurring issue for this [set of authors] . ***  , 
 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 two indicated sub-Figures in the Figure 1, namely the Figure 1a and 1c, which are the graphic exhibition of the migration assay and invasion assay conducted using OL and Pol cells, respectively. We agree with the assessment that (1) the top panel of Figure 1a showed 3 representative microscopic views of the migration results of OL cells of which image 1 and image 2 contain overlapping areas; (2) the 1st image one of lower panel of Figure 1a (POL migration) is identical to that of the 2nd image of the lower panel of Figure 1c (POL invasion). We are grateful for that you alert us of such errors present in the Figure 1. 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. The migration and invasion assay were repeated 3 times. For each repeat, at least 3 experimental repeats were set up for each experimental group (in this case, the OL and POL cells). For visualization and quantification of the two assays, the membranes of the transwell that contained the migrated or invaded cells were fixed, stained and mounted to a slide. Cells that migrated or invaded to the lower side of the membrane were photographed. For the quantification, we routinely take the images of 10 randomly chosen view fields for each slide (the Transwell membrane will be mounted onto a slide for microscopic observation and quantification). We found the original lab records in the lab note book that summarized the migration and the invasion assay of the OL and POL cells and the figures made for publication (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: For Transwell invasion assays, Line2: matrigel:DMEM without FBS=1:8, to dilute matrigel Line3: add 30ul diluted matrigel to Transwell, put Transwell in 37u00b0C incubator to dry the Transwell Line5: diluted Matrigel resembles the extracellular matrix in vivo Line6: Experimental Date: April 10th, 2018 Line7: Quantitative statistics of migration and invasion experiment results POL OL Line8: Staff: Zhiwei Sun Line14: migration ability: POL has stronger migration ability than OL (below the figure of migration assay) Line20: invasion ability: POL has stronger invasion ability than OL(below the figure of migration assay)  Our observations and responses based on the examination of the original lab records: (1)tThe overlapping areas in the two images: The printed figure summarizes the migration assay in the lab records is the same as Figure 1a in the published paper. Thus, the images 1 and 2 of the top panel of OL migration assay also contain the overlapping areas as indicated. The overlap area in the image 1 and image 2 of the top panel of Figure 1a occurred during image taking. It is most likely that we took the images of two adjacent view fields that had overlapped area. We happened to choose two representative images with overlapping view fields without even noticing (the printed figure of the migration assay is identical to Figure 1a in the paper) when figures were made for publication. Thus, there is no intentional manipulation of the images. (2)tIdentical images in Figure 1a and Figure 1c: The printed figure summarizes the invasion assay in the lab records is identical to that in Figure 1c in the published paper. Thus, the error occurred and documented in the lab book. Based on the examination and analysis, we made a careless error during the making of the summarized figure for publication. We chose the same image when we made the two sub-lower panels of Figure 1a and 1c. We failed to identify the error during reviewing of the manuscript by all authors. 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 sending 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 1 based on the original data for your examination. The revised figure:  We sincerely apologize for our careless that resulted in the unintentional errors present in the published paper. We will learn from this lesson so that it will not happen again. We thank you for pointing the errors out and giving us an opportunity to correct. We will contact the Journal to make the correction as soon as possible. , 
 #2 First of all, we examined the two indicated images in the mid-panel Figure 4c, which is the representative graphic exhibition (with three images) of the invasion assay conducted using OL-siRNA-Sec23a cells. We agree with the assessment that the image 1 and image 3 contain a small overlapping area. 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. The invasion assay was repeated 3 times. For each repeat, at least 3 experimental repeats were set up for each experimental group. For the visualization and quantification of the two assays, the membranes of the transwell that contained the migrated or invaded cells were fixed, stained and mounted to a slide. Cells that invaded to the lower side of the membrane were photographed. For the quantification, we routinely take the images of 10 randomly chosen view fields for each slide. We found the original lab records in the lab note book that summarized the study and the figures made for publication (see the inserted photocopy of the lab records). Since one out of the two pages of the lab notes was written in Chinese, we made a line-by-line translation of the lab note for your Translated version: Line10: Date: September 8th, 2018 Line11: Content: Ebiomedicine-manuscript Line12: Location: room 310, Life Sciences Institute Line13: Staff: Zhiwei Sun comprehension and review. Our observations and responses based on the examination of the original lab records: (1)The over lapping areas in the two images: The printed figure summarizes the invasion assay in the lab records is the same as Figure 4C in the published paper. Thus, the images 1 and 3 of the mid-panel of the invasion assay also contain the overlapping areas as indicated. Thus, the overlap area in the image 1 and image 3 of the mid panel of Figure 4c occurred during image taking. It is most likely that we took the images of two adjacent view fields that had overlapped area. We happened to choose two representative images with overlapping view fields without even noticing (the printed figure of the invasion assay is identical to Figure 1a in the paper) when figures were made for publication. Thus, there is no intentional manipulation of the images.   We sincerely apologize for our careless that resulted in the unintentional errors present in the published paper. We will learn from this lesson so that it will not happen again. We thank you for pointing the errors out and giving us an opportunity to correct. We will contact the Journal to make the correction as soon as possible. We also provide the revised figure based on the original data and our original experiment records for your reference. The revised figure:  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 4 based on the original data for your examination. , 
 Corrigendum. August 1, 2021  The authors (sic) have recently noticed that the published version of this article contained errors in Fig. 1 and Fig. 4. Unintentional overlaps and duplications were found in the representative images of migration and invasion assays. These inadvertent errors were made during figure preparation. The corrected Fig. 1 and Fig. 4 are presented below. These corrections do not change the description, interpretation, or the original conclusions of the manuscript. The authors apologize for any inconvenience caused by these unintentional errors. ***   ,