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©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
Effects of targeted-edited oncogenic insulin-like growth factor-1 receptor with specific-sgRNA on biological behaviors of HepG2 cells
Min Yao, Yin Cai, Zhi-Jun Wu, Ping Zhou, Wen-Li Sai, De-Feng Wang, Li Wang, Deng-Fu Yao
Min Yao, Wen-Li Sai, De-Feng Wang, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China
Min Yao, Ping Zhou, Department of Medical Immunology, Medical School of Nantong University, Nantong 226001, Jiangsu Province, China
Yin Cai, Department of Oncology, Xinghua People’s Hospital, Xinghua 225700, Jiangsu Province, China
Zhi-Jun Wu, Department of Oncology, Affiliated Nantong Rehabilitation Hospital of Nantong University, Nantong 226002, Jiangsu Province, China
Li Wang, Research Center for Intelligent Information Technology, Nantong University, Nantong 226019, Jiangsu Province, China
Author contributions: Yao M, Cai Y, Wu ZJ and Zhou P contributed equally to this work and wrote the first draft; Zhou P and Wang L contributed to the methodology, data curation, and formal analysis; Sai WL and Wang DF analyzed and wrote the manuscript; Wang L and Yao DF revised the manuscript; All authors approved the final version of the manuscript.
Supported by Projects of the National Natural Science Foundation of China, No. 81873915, No. 31872738 and No. 81673241; Key Plan of Nantong S & T Development, No. MS12020021; and Program of Medical School S & T of Nantong University, No. 2018YFC0116902.
Institutional review board statement: This study was approved by the Ethics Committee of the Affiliated Hospital of Nantong University (TDFY2013008), China.
Informed consent statement: All study participants, or their legal guardian, provided informed written consent prior to study enrollment.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https:// creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Deng-Fu Yao, MD, PhD, Full Professor, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, No. 20 West Temple Road, Nantong 226001, Jiangsu Province, China. yaodf@ahnmc.com
Received: December 13, 2021
Peer-review started: December 13, 2021
First decision: April 16, 2022
Revised: April 28, 2022
Accepted: August 25, 2022
Article in press: August 25, 2022
Published online: October 6, 2022
BACKGROUND
Insulin-like growth factor-1 receptor (IGF-1R) is over-expressed in hepatocellular carcinoma (HCC). However, the relationship between IGF-1R activation and HCC progression remains unidentified.
AIM
To investigate the effects of editing IGF-1R on the biological features of HCC cells.
METHODS
Immunohistochemistry analyzed the expressions of IGF-1R and P-glyco protein (P-gp) in HCC tissues and their distal non-cancerous tissues (non-Ca). IGF-1R was edited with Crispr/Cas9 system, screened specific sgRNAs, and then transfected into HepG2 cells. CCK-8, scratch wound test detected cell proliferation, migration, invasion and transwell assays, respectively. Alterations of IGF-1R and P-gp were confirmed by Western blotting. Alterations of anti-cancer drug IC50 values were analyzed at the cell level.
RESULTS
The positive rates of IGF-1R (93.6%, χ2 = 63.947) or P-gp (88.2%, χ2 = 58.448) were significantly higher (P < 0.001) in the HCC group than those (36.6% in IGF-1R or 26.9% in P-gp) in the non-Ca group. They were positively correlated between high IGF-1R and P-gp expression, and they were associated with hepatitis B virus infection and vascular invasion of HCC. Abnormal expressions of circulating IGF-1R and P-gp were confirmed and associated with HCC progression. Biological feature alterations of HCC cells transfected with specific sgRNA showed IGF-1R expression down-regulation, cell proliferation inhibition, cell invasion or migration potential decreasing, and enhancing susceptibility of HepG2 cells to anti-cancer drugs.
CONCLUSION
Edited oncogenic IGF-1R was useful to inhibit biological behaviors of HepG2 cells.
Core Tip: Abnormal expression of insulin-like growth factor-1 receptor (IGF-1R) was associated with hepatocellular carcinoma (HCC). IGF-1R level was significantly higher in HCC more than that in their noncancerous tissues. Circulating IGF-1R continued to increase from benign liver disease to HCC. Down-regulating expression of IGF-1R with a specific sgRNA was markedly affected on biological behaviors of HCC cells, including inhibiting cell proliferation, decreasing cell migration or invasion potential, increasing cell apoptosis and enhancing cell susceptibility to anti-tumor drugs and indicated that oncogenic IGF-1R should be a promising targeted-molecule for HCC therapy.
