25-Hydroxycholesterol promotes migration and invasion of lung adenocarcinoma cells
Authors: Yinping Lv
Li Chen
Yanliang Lin
Yibing Wang
Guozhe Xian
Lishan Zhang
Journal: Biochemical and Biophysical Research Communications (2017)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: 25-Hydroxycholesterol; IL-1β; Invasion; LXR; Lung adenocarcinoma
Affiliations: Center Laboratory, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China., Department of Hand and Foot Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China., Department of Hepatobiliary Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China., Department of Medical Engineering, Jinan 250021, China., Center Laboratory, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China. Electronic address: ., Department of Burn and Plastic Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China. Electronic address: .

Abstract:  25-hydroxycholesterol (25-HC) is enzymatically produced by cholesterol 25-hydorxylase in various organs and is involved in many processes, including lipid metabolism, inflammation and the immune response. However, the role of 25-HC in the migration and invasion of lung adenocarcinoma (ADC) cells remains largely unknown. In this study, we demonstrated that 0.1 μM 25-HC promoted ADC cell migration and invasion without affecting cell proliferation, especially after coculture with THP1-derived macrophages. Further investigation showed that 0.1 μM 25-HC significantly stimulated interleukin-1β (IL-1β) secretion in a coculture system and increased the expression of LXR and Snail. IL-1β also mimicked the effect of 25-HC. LXR knockdown notably blocked the 25-HC-induced Snail expression, migration and invasion in both the monoculture system and the coculture system, but it did not impact the effect of IL-1β, which suggested that IL-1β functioned in an LXR-independent manner. These results suggested that 25-HC promoted ADC cell migration and invasion in an LXR-dependent manner in the monoculture system but that in the coculture system, the 25-HC-induced IL-1β secretion enhanced the effect of 25-HC in an LXR-independent manner. 

Retracted: No

PubPeer comments:
 Figure 1C. The 1.0 uM panel in the top row appears to share some similarities with the 10 uM panel in the bottom row. ![file] From the Acknowledgments:  This work was supported by the National Natural Science Foundation of China (Grant No. 81572272; 81201778; 81201471; 81171818), the Promotive Research Fund for Excellent Young and Middle-aged Scientists of Shandong Province (Grant No. BS2011SW036; BS2013YY065), the Foundation for Outstanding Young Scientist in Shandong Province (Grant No. BS2013YY058) and the Science and Technology Development Plan Project of Shandong Province (Grant No. 2014GSF118157). , 
 Figure 2A. * Yellow boxes: Two of the 0h panels appear to overlap with each other. ![file] , 
 Due to the large number of experimental pictures and our negligence, we may put the wrong picture, we will contact the magazine to modify, thank you! , 
 #1 ![file] ![file] Dear Elisabeth M Bik We have checked the original images we exported from system and the results are showed above, Thank you again! As for Figure 2A, because scratch assay at 0h, the cells did not be treated with 25-HC, all the scratches are similar, for which the image at 0h in 25-HC-treated group was mistaken, Although it did not affect the real results. Thank you for you reminding! , 
 #4 Is this area of repetition expected? Perhaps Dr. Bik can also review and comment. ![file] ![file] , 
 #5 Thank You! We have noticed this. We will contact the magazine to modify, thank you! , 
 Thank you dr. Lin for checking and finding the original files. The files you showed here indeed correspond to the files as published in the paper. As such, the repetitions I marked above in #1 are still visible. Posting the same image here does not take away any of my initial concerns, I am afraid. I cannot think of any reason how two different samples could end up sharing some groups of dots but not other groups of dots. Even if the machine was not cleaned well or if two files would be super imposed, that could not explain for similarities of data points between two graphs in parts of the graphs but not in others. Here are the two panels again, marked as in #1, with some additional findings. The similarities are quite striking. I am not sure how if the journal can modify, since these are basically the same images with the same problems. ![file] , 
 #7 Thanks for Dr Elisabeth M Bik. We analyzed possible reasons. We think that the two groups of cells share a single test tube, which is not completely cleaned. We are so sorry for bothering you. We have contacted the magazine to resubmit another set of experimental results, and we will show you as soon as possible after I have prepared them. Thank you again! , 
 Dr. Lin, if you would run the same sample from the same test tube twice on the same FACS machine, it would still not give identical dots. The FACS machine would suck up different cells and the dots would land on roughly the same but never exactly the same spot. If I would put another sample in a dirty tube, it would definitely not results in the same dots. I d be happy to ask one of my Twitter followers to explain to you how flow cytometry works, and how your theory does not make much sense. , 
 The authors regret that Fig. 1C (H1975 cells stimulated with 10 u03bcM 25-HC) and Fig. 2A (A549 cells stimulated with 25-HC at 0 h in monoculture) were mistakenly uploaded in the original publication. In order to ensure that the results are from the same experiment, the authors have revised Fig. 1C by placing parallel data, and included corrected Fig. 2A. These corrections do not affect the overall conclusion in the manuscript. The authors would like to apologize for any inconvenience caused. ![file] ![file] , 
 Unfortunately, although the authors can be congratulated on persuading the journal to correct the figures above, their work is not yet finished. Images in Figure 2B appear to overlap, including one image that seems to have been rotated and resized, suggesting this is more serious than merely an error during figure assembly. *** ![file] , 
 28 May 2021 ![file] ,