GATA3 suppresses human fibroblasts-induced metastasis of clear cell renal cell carcinoma via an anti-IL6/STAT3 mechanism
Authors: Renfang Xu
Xiaozhou He
Hao Lu
Ying Xia
Qianqian Shi
Guanglai Song
Dong Xue
Journal: Cancer Gene Therapy (2020)
Discipline: CLINICAL MEDICINE
Keywords: Tumorigenesis; Metastasis; Clear-cell renal cell carcinoma; Fibroblasts; Signaling pathways
Affiliations: The Third Affiliated Hospital of Soochow University, Changzhou, 213000, China., The Third Affiliated Hospital of Soochow University, Changzhou, 213000, China. ., Shanghai Key Laboratory of Acupuncture Mechanism and Acupoint Function, Fudan University, Shanghai, 200433, China. .

Abstract:  Tumorigenesis and metastasis depend on intricate interactions between genetically altered tumor cells and their surrounding microenvironment. It is, however, unclear regarding the molecular mechanisms underlying the progress and metastasis of human clear-cell renal cell carcinoma in the microenvironment with fibroblasts. In this work, we investigated the effect of normal fibroblasts on the metastasis of renal cancer and the relevant signaling pathways. We isolated normal fibroblasts from normal renal tissues and used normal fibroblast-conditioned medium culture renal cancer cells. The CCK-8 and transwell assays showed that normal fibroblasts conditioned medium significantly enhanced ccRCC cell migration. IL6 mediated the cross talk between normal fibroblasts and the cancer cells, and promoted tumor cell migration through the STAT3 pathway. In contrast, GATA3 was downregulated at both mRNA and protein levels in the normal fibroblast-conditioned medium treated with renal cancer cells, but upregulated in adjacent normal tissues. GATA3 overexpression significantly reduced STAT3 phosphorylation and attenuated the migration in both renal cancer cell and IL6-stimulated renal cancer cell. Taken together, our findings suggest that the IL6/STAT3 pathway plays a crucial role in the normal fibroblast-enhanced clear-cell renal cell carcinoma metastasis, while GATA3 may mitigate this effect by inhibiting IL6/STAT3 signaling. 

Retracted: No

PubPeer comments:
 Figure 2B has duplicated images with different fields of view: ![file] Figure 6A appears to have western blot images spliced together: ![file] , 
 We highly appreciate your valuable comments! We have rechecked the original data. The first author, Shi QQ, found that she made the low-level mistake in Fig. 2B, as you indicated, when selecting representative images from countless image data. We apologize for it! We have contacted the journal and submitted a corrigendum. As for Fig. 6A, u201c769-Pu201d and u201cACHNu201d were indeed from different experiments and there was no relationship between them in this figure. To avoid potential misleading, we have separated them in a revised figure and included it in the corrigendum. Again, we thank you very much for your reading and comments. Really appreciate it! , 
 #2 Regarding 6A, the GAPDH bands do *not* show evidence that they were cropped. Could you please provide high resolution *uncropped* original images to reassure readers? , 
 #3 Thank you very much for your comments. I have rechecked all our data and realized that I made the mistakes when choosing representative images from our countless images. Itu2019s solely my responsibility and I sincerely apology for it. We will correct the errors with the journal. The corrections do not change our statistical analysis and conclusion of the article. Per your request, here I attach uncropped images of ACHN from 3 experiments. As you can see, GATA3 signal densities largely increased after the transfection with the GATA3 overexpression plasmid, while GAPDH signal bands had no appreciable changes. ![file] , 
 November 6, 2020 Correction to: Cancer Gene Therapy https: doi.org 10.1038 s41417-019-0146-2 The original version of this Article contained misplaced mislabeled images in Figs. 2b, 6a and 7b. All these figures have been corrected in both the PDF and HTML versions of the Article. ![file] ![file] ![file] Figure 2b: The wrong images were placed in the original when choosing representative images from the countless image data. The figure has now been corrected. Figure 6a: Parts of the image were labelled incorrectly. The correct group names should be u201c769-P, ACHN, 786-O and Caki-1u201d as stated in the legend of Fig. 6. Moreover, we have separated these four groups of images to avoid potentially misleading readers, since there was no comparison among the data of these groups. Figure 7b: Mismatched total STAT3 and GAPDH images. Although they had no appreciable changes in all groups and nobody may notice it, we feel that the correction will be better in terms of scientific spirit. The amendments to the above mentioned figures do not affect the statistical analysis and conclusions of the article. For example, in Fig. 6a, GATA3 protein density of ACHN largely increased in the cells after the transfection with the GATA3 overexpression plasmid, while GAPDH signal bands had no appreciable changes. We have provided the original and uncropped images from three different experiments strongly support this conclusion. , 
 Figure 2d and 4a appear to have an overlap, after resizing and recoloring. The resolution appears different as well. It is difficult to believe that this, when considered alongside the previous issues addressed in the correction, can be due to mistakes during figure assembly. I have shared my concerns with the journal. *** Closeup ![file] , 
 #6 We have rechecked the figures and found no errors at all. Indeed, the images indicated by the reader were from the same group, but for different purposes with different view fields. In Figure 2B, we showed the effects of five different NF-CMs on the migration of ccRCC cells. All of them, including NF171006, enhanced the 769-P cell migration. In Figure 4A, we showed the effect of NF171006, one of the five NF-CMs, on the 769-P cells with without anti-IL6. Since all these NF-CMs had the similar effect on the migration, we simply labeled u201cNF-CMu201d here to better reflect the meaning of the outcome results. We apologize for the misunderstanding we might cause. We thank the readers for his her careful reading of our paper and will appreciate if he she could clarify it with the journal since he she shared this issue with the journal. , 
 ![file] , 
 #8 Dear Reader, tThank you for reading our article. tPlease see our correct version published on Nov 6, 2020 . tAs for the images in Figures 2B and 4A indicated in pink, they were indeed from the same group with different view fields for different purposes. In Figure 2B, we showed the effects of five different NF-CMs on the migration of ccRCC cells. All of them, including NF171006, enhanced the 769-P cell migration. In Figure 4A, we showed the effect of NF171006, one of the five NF-CMs, on the 769-P cells with without anti-IL6. Since all the NF-CMs had the similar effect on the migration, we simply labeled u201cNF-CMu201d here to better reflect the meaning of the outcome results. We replied the same question from another reader in April 2021. With best regards. Qianqian Shi , 
 #8 Dear Reader, tThank you for reading our article. tPlease see our correct version published on Nov 6, 2020 . tAs for the images in Figures 2B and 4A indicated in pink, they were indeed from the same group with different view fields for different purposes. In Figure 2B, we showed the effects of five different NF-CMs on the migration of ccRCC cells. All of them, including NF171006, enhanced the 769-P cell migration. In Figure 4A, we showed the effect of NF171006, one of the five NF-CMs, on the 769-P cells with without anti-IL6. Since all the NF-CMs had the similar effect on the migration, we simply labeled u201cNF-CMu201d here to better reflect the meaning of the outcome results. We replied the same question from another reader in April 2021. With best regards. Qianqian Shi ,