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.

AIM: To investigate the function and the mechanism of transplanting bone marrow derived peripheral blood mesenchymal stem cells(PBMSCs) on restenosis after carotid balloon angioplasty in the model of carotid atherosclerosis rabbits,and to determine if the functions of PBMSCs are enhanced after hypoxia preconditioning.METHODS: Bone marrow cells were mobilized by granulocyte colony-stimulating factor(G-CSF),and PBMSCs were collected through density gradient centrifugation and adherent culture,labeled with enhancement type green fluorescent protein(EGFP) genes.All animals with carotid atherosclerosis stenosis were randomly divided into three groups: hypoxia preconditioning group(n=24,received intravenous transplantation of PBMSCs with hypoxia preconditioning),non-hypoxia preconditioning group(n=24,received normal culture of PBMSCs) and control group(n=24,only received equal-volume of culture medium).Vascular endothelial growth factor(VEGF) was determined by enzyme linked immunosorbent assay(ELISA) at 7 d,14 d and 28 d post-angioplasty,respectively.The vessel morphology,the homing of MSCs and the reendothelialization were analyzed with Weigert staining and immunohistochemistry.RESULTS: Compared to control group,the level of VEGF significantly increased in both hypoxia preconditioning group and non-hypoxia preconditioning group at all time points(P