Clinical and Translational Research
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Clin Cases. Oct 6, 2022; 10(28): 10017-10030
Published online Oct 6, 2022. doi: 10.12998/wjcc.v10.i28.10017
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
Processing time: 288 Days and 10.5 Hours
ARTICLE HIGHLIGHTS
Research background

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death with a high incidence and mortality rate in China. Insulin-like growth factor 1 receptor (IGF-1R) signaling triggers cell proliferation, liver growth and tissue regeneration during embryonic development. Unbalanced IGF-1R signaling can promote HCC cell proliferation and regulating IGF-1R gene transcription should be useful as a potential therapy targeting HCC.

Research motivation

IGF-1R as key signaling of the IGF axis in HCC progression was investigated in sera or tissues from HBV-related HCC patients. We analyzed the relationship between IGF-1R and multi-drug resistance (MDR) and edited the IGF-1R gene for downing-regulating expression to confirm effects on proliferation and a potential therapeutic role for HCC cells.

Research objectives

The expressing statues and clinicopathological characteristics of IGF-1R or P-glyco protein (P-gp) were investigated in the circulating blood and tissues of HCC patients and editing IGF-1R gene at the mRNA transcription level to observe effects on biological behaviors HepG2 cells and their synergistic role with anti-cancer drugs on reversal MDR of HCC.

Research methods

Comparative analysis of IGF-1R and P-gp expression in tissues or sera of HCC patients were analyzed by immunohistochemistry and confirmed by Western blotting. Specific sgRNA was screened among editing IGF-1R gene with Crispr/Cas9 system and then transfected into HepG2 cells. CCK-8, scratch wound test detected HCC cell proliferation, migration, invasion and transwell assay, respectively.

Research results

Abnormal over-expression of IGF-1R and P-gp were confirmed in tissues or sera of HCC patients with a positive close correlation between IGF-1R and P-gp and related to HBV infection or vascular invasion during HCC progression. HepG2 cell biological features were altered by specific IGF-1R-sgRNA with down-regulation, cell proliferation inhibition, cell invasion or migration potential decreasing and enhancing cell susceptibility to anti-cancer drugs.

Research conclusions

Based on this these studies, high IGF-1R or P-gp expression has been confirmed related to the progression or therapeutic effect of HCC. Although the accurate mechanism for IGF-1R reactivation in HCC remains to be explored, specific edited oncogenic IGF-1R gene is promising for inhibiting proliferation, altering biological features or as potential modulators for reversal MDR of HCC cells.

Research perspectives

Abnormal expression of hepatic IGF-1R level was associated with HCC progression. Inhibiting IGF-1R expression could markedly affect the biological behaviors of HCC cell proliferation, migration or invasion, cell apoptosis and drug susceptibility suggesting that the IGF-1R gene could be a promising targeted molecule for HCC therapy.