关键词: 胃癌, HO-1基因, 干扰载体, SGC-7901细胞

Abstract: Objective To investigate the effect and function of HO1 gene in its human gastric cancer SGC-7901 cell line by constructing the eukaryotic siRNA expression vector of HO1 gene and to explore its potential role in gastric carcinogenesis. Methods A recombinant plasmid carrying siRNA specific for HO1 gene expression vector was constructed and transfected into SGC-7901 cells by lipofectin-mediated method. Cells (SGC-7901-pS/HO1) stably expressing siRNA specific for HO-1 gene were selected using G418. SGC-7901 cells untransfected and those transfected with empty pS plasmid (SGC-7901-pS) were used as controls. The expression levels of HO1 gene were detected by real-time PCR and Western bloting in stably transfected-SGC-7901(SGC-7901-pS/HO1) cells, biological function of HO1 gene was analyzed by cell growth activity and soft agar colony formation assay. Results Compared with SGC-7901-PS cells, the expression of HO1 protein of SGC-7901-pS/HO1 cells was an average 5.58-fold decrease(0.321±0.051 vs 1.675±0.153, P <0.05). The colony formation rates of SGC-7901-pS/HO1 cells were significantly reduced compared to that of control group (p < 0.05). The cell growth activity showed same tendency in SGC-7901-pS/HO1 group, SGC-7901-pS cells group and SGC-7901 cells group. Conclusion the eukaryotic expression vector of siRNA specific for HO1 gene was constructed successfully,which might contribute to probe the functions of HO1 in human gastric cancer in depth.

Key words: Gastric carcinoma, HO-1 gene, siRNA vector, SGC-7901 cells