摘要
目的:构建CD147-shRNA重组质粒并检测其对胃癌细胞SGC7901内源性CD147表达的抑制作用。方法:构建靶向CD147的siRNA表达载体pSilencer-CD147siRNA,稳定转染至SGC7901细胞中,通过Real-time PCR和Western blot法检测转染细胞中CD147表达水平的变化。结果:酶切鉴定和测序证实成功构建pSilencer-siRNA1,pSilencer-siRNA2和pSilencer-negative,转染后SGC7901中CD147表达水平显著降,而以pSilencer-siRNA2抑制作用最为显著。结论:构建pSilencer-CD147siRNA成功,并筛选出基因抑制效果抑制最佳的pSilencer-siRNA2,为进一步实验打下了基础。
Objective： To construct recombinant plasmid CD147-shRNA and detect its inhibitory effect on CD147 expression of gastric cancer cell line SGC7901. Methods： Small interfering RNA （siRNA） expressing vectors targeting CD147 were transfected into human gastric cancer cells SGC7901 and CDl47 expression was monitored by real-time PCR and Western blot. Results： CD1 47-shRNAs recombinant plasmid was completely and correctly constructed confirmed by restriction enzyme digestion and sequencing. All the plasmids had gene silencing effect on CD147 expression of SGC7901, but the inhibit effect of pSilencer-siRNA2 was most. significant. Conclusion： We successfully constructed pSilencer-CD147siRNA and screened out pSilencer-siRNA2 which has the best gene silence effect.
出处
《现代生物医学进展》
CAS
2010年第22期4204-4208,共5页
Progress in Modern Biomedicine
基金
国家自然科学基金资助项目(30873022)
关键词
RNA干扰
CD147
载体构建
胃癌
RNA interference
CD147
Vector construction
Gastric cancer
