The RNA-binding protein GRSF1 promotes hepatocarcinogenesis via competitively binding to YY1 mRNA with miR-30e-5p
Authors: Dongdong Tong
Chen Huang
Xiaofei Wang
Lili Han
Journal: Journal of Experimental & Clinical Cancer Research (2022)
Discipline: CLINICAL MEDICINE
Keywords: GRSF1; HCC; RBP; YY1; miR-30e-5p.
Affiliations: Department of Oncology, The Second Affiliated Hospital, College of Medicine, Xi'an Jiaotong University, Xi'an, 710004, Shaanxi, China. ., Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education of China, Xi'an Jiaotong University, No.277 Yanta West Road, Xi'an, 710061, Shaanxi Province, China.

Abstract:  Background: Dysregulation of RNA binding protein (RBP) expression has been confirmed to be causally linked with tumorigenesis. The detailed biological effect and underlying mechanisms of the RBP GRSF1 in hepatocellular carcinoma (HCC) remain unclear. Methods: HCC cells with stable knockdown of GRSF1 were established using two sh-RNA-encoding lentiviruses. The functions of GRSF1 in HCC were explored using MTT, colony formation, flow cytometry, and Transwell assays and a xenograft model. Transcriptomic sequencing in GRSF1-deficient MHCC-97H cells was carried out to identify the downstream effector of GRSF1. The regulatory mechanisms among GRSF1, YY1 and miR-30e-5p were investigated via RNA immunoprecipitation, luciferase, RNA pull-down and ChIP assays. Several in vivo assays were used to assess the selectivity of the small-molecule compound VE-821 in HCC and to confirm the absence of general toxicity in animal models. Results: GRSF1 was frequently increased in HCC tissue and cells and was associated with worse clinical outcomes. GRSF1 functions as a novel oncogenic RBP by enhancing YY1 mRNA stability, and the GUUU motifs within the YY1 3`UTR 2663-2847 were the specific binding motifs for GRSF1. YY1 feedback promoted GRSF1 expression by binding to the GRSF1 promoter. In addition, YY1 was a critical target of miR-30e-5p, which was confirmed in this study to inhibit HCC hepatocarcinogenesis. GRSF1 and miR-30e-5p competitively regulated YY1 by binding to its 3`UTR 2663-2847 region. Finally, we identified that VE-821 blocked HCC progression by inhibiting the GRSF1/YY1 pathway. Conclusion: This study revealed the interaction network among GRSF1, YY1 and miR-30e-5p, providing new insight into HCC pathogenesis, and indicated that VE821 may serve as a novel agent with potential for HCC treatment through inhibition of the GRSF1/YY1 axis. 

Retracted: No

PubPeer comments:
 Images in Figure 4I (left) and 7D (right) appear to overlap, after rotation. *** ![file] , 
 #1 Thank you for the comments, we apologize for the carelessness in the preparation of the data. We thank you for finding this errors. Actually, one of the image of Transwell assay in figure7D is misused. I repeatedly confirmed the data stored on the computer and the experimental operation records on my own notebook, and found the accurate data . We have checked the original result as soon as possible and corrected them. Please see the attachment for details. If you have any questions, please do not hesitate to contact me. Thank you again! ![file] , 
 #2 Thank you for responding. For clarity: will you be contacting the joy to publish a correction? , 
 #3 We are trying, but since we have no experience, I don t know whether it can work. If not, we may take the initiative to withdraw the manuscript sincerely. After all, it is really my mistake. We re not serious enough. Thank you so much. ,