[ Knock-out Affectsthe Proliferation, Migration and Invasion Ability of Hepatocellular Carcinoma HepG2 Cell Line ].

Objective: To investigate the effect of nuclear receptor knockout on proliferation and migration ability of human hepatocellular carcinoma cell line HepG2.

Methods: -The gene knockout HepG2 cell line was abtained by CRISPR/Cas9 genome editing technique with specific DNA modification of the target gene. The gene targeting vectors were co-transfected into HepG2 cells. Through cloning and screening, the gene knockout HepG2 cell line was constructed, PCR, sequencing and Western blot methods were carried out for the identification of the gene knockout HepG2 cell line. The expression level of tumor migration and invasion-associated gene in gene knockout cell was determined by real-time quantitative PCR (qRT-PCR) and was compared with normal cell as control.MTT, cell scratch and Transwell experiments were conducted in order to explore the effect of gene on HepG2 cell's ability of proliferation, migration and invasion.

Results: A gene knockout monoclonal cell line, which was identified by PCR, sequencing and Western blot, was successfully constructed and named HepG2 C5 ( ). qRT-PCR results showed that knockout resulted in up-regulation of matrix metalloproteinase-2 (), matrix metalloproteinase-9 and extracellular matrix protein-1 () gene expression ( < 0.05) and down-regulation of E-cadherin () gene expression (=0.05).Results of MTT, cell scratch and transwell experiments showed that HepG2 C5 had stronger proliferation, migration and invasion ability than control cells ( < 0.05).

Conclusion: gene knockout could change the expression of migration and adhesion-associated genes in HepG2 cell, and then affect the proliferation, migration and invasion ability of HepG2 cells.