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

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

World J Clin Cases. 2022 Oct 6;10(28):10017-10030. doi: 10.12998/wjcc.v10.i28.10017.

Authors

Min Yao^{1

2}, Yin Cai³, Zhi-Jun Wu⁴, Ping Zhou², Wen-Li Sai¹, De-Feng Wang¹, Li Wang⁵, Deng-Fu Yao⁶

Affiliations

¹ Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
² Department of Medical Immunology, Medical School of Nantong University, Nantong 226001, Jiangsu Province, China.
³ Department of Oncology, Xinghua People's Hospital, Xinghua 225700, Jiangsu Province, China.
⁴ Department of Oncology, Affiliated Nantong Rehabilitation Hospital of Nantong University, Nantong 226002, Jiangsu Province, China.
⁵ Research Center for Intelligent Information Technology, Nantong University, Nantong 226019, Jiangsu Province, China.
⁶ Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China. yaodf@ahnmc.com.

Abstract

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%, χ ² = 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.

Keywords: Biological behaviors; Growth inhibition; Hepatocellular carcinoma; Insulin-like growth factor-1 receptor; Multidrug resistance; Synergistic effects.