Role of B7-H6 over-expression in NK cell-mediated apoptosis of hepato-cytes
10.3969/j.issn.1000-4718.2017.11.028
- VernacularTitle:过表达B7-H6在自然杀伤细胞介导的肝细胞凋亡中的作用
- Author:
Yong ZOU
1
;
sheng Zhe LIN
;
chan Yu CHEN
;
hong Si LIAO
;
Qing YUAN
;
jun Dong LIN
Author Information
1. 中山大学附属第三医院
- Keywords:
B7 homologue 6;
Eukaryotic expression vector;
Natural killer cells;
Hepatocytes;
Apoptosis
- From:
Chinese Journal of Pathophysiology
2017;33(11):2095-2098
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the role of B7 homologue 6 (B7-H6) over-expression in natural killer (NK) cell-mediated hepatocyte apoptosis. METHODS:The full-length fragment of B7-H6 gene was amplified by PCR and sub-cloned into linearized eukaryotic expression vector pIRES2-EGFP to construct recombinant B7-H6 over-expression vector pIRES2-EGFP-B7-H6. The recombinant plasmid was identified by double digestion, PCR and sequencing, and was then transfected into L02 cells. The expression of EGFP was observed by fluorescence microscopy and the transfection efficiency was evaluated by flow cytometry. B7-H6 expression was confirmed by qRT-PCR and Western blot. The L02 cells transfect-ed with pIRES2-EGFP-B7-H6 recombinant plasmid were co-cultured with NK-92 cells at different effector/target ratios,and the cytotoxicity of NK-92 cells was evaluated by CCK-8 assay.RESULTS:The strong green fluorescence in the L02 cells was observed under fluorescence microscope 48 h after transfection. The transfection efficiency reached 92.6%. The ex-pression of B7-H6 at mRNA and protein levels was remarkably increased 48 h after transfection. The cytotoxicity of NK-92 cells against L02 cells transfected with pIRES2-EGFP-B7-H6 plasmid was significantly higher than that of the null vector transfection group (P<0.05).CONCLUSION:The recombinant eukaryotic expression vector pIRES2-EGFP-B7-H6 was constructed successfully. The cytotoxic effect of NK-92 cells against L02 cells can be enhanced by transfecting L02 cells with pIRES2-EGFP-B7-H6 plasmid.
