ABSTRACT
Objective To investigate the role of non-small cell lung cancer metastasis-related Trim72 gene in hepatocellular carcinoma (HCC) using clustered regularly interspaced short palindromic repeats (CRISPR) Cas9 system.Methods Hepa1-6 (Cas9) HCC cells were established with stable expression of Cas9 protein,and then specific gene knockout was performed using sgRNA targeting Trim72 gene.After obtaining the Hepa1-6 (Trim72-KO) cells,the metastasis and invasion abilities of the cells were evaluated by in vitro Transwell assay and in vivo subcutaneous lung metastasis examination.Results Hepa1-6 (Trim72-KO) cell line was successfully established by the CRISPR-Cas9 system.Transwell assay indicated that the mobility of Hepa1-6 (Trim72-KO) cells was increased compared to the control cells.Transwell assay indicated that the metastasis and invasion of Hepa1-6 (Cas9) HCC cells were enhanced after the knockout of Trim72 gene.The pathological examination of lung metastasis of subcutaneous tumor in vivo showed that the subcutaneously metastatic ability of Hepa1-6 (Trim72-KO) cells (the experimental group) was significantly stronger than Hepa1-6 (Cas9) cells (the control troup) that were not transferred to the corresponding sgRNA.Conclusions The trim72 gene knocked-out HCC cells were obtained by CRISPR-Cas9 system,which showed stronger metastasis and invasion abilities than the control cells.It is suggested that Trim72 gene may play an important role in the invasion and metastasis of HCC,and Trim 72 gene is expected to be a potential target for gene therapy of liver cancer.