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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 CD147-shRNA recombinant plasmid and test its inhibitory effect on the expression of endogenous CD147 in gastric cancer cell line SGC7901. Methods: The pSilencer-CD147 siRNA targeting CD147 was constructed and transfected into SGC7901 cells. The expression of CD147 in transfected cells was detected by Real-time PCR and Western blot. Results: The pSilencer-siRNA1, pSilencer-siRNA2 and pSilencer-negative were successfully constructed by restriction enzyme digestion and sequencing. The expression of CD147 in SGC7901 transfected cells was significantly decreased, while pSilencer-siRNA2 was the most effective inhibitor. Conclusion: The successful construction of pSilencer-CD147 siRNA and the screening of pSilencer-siRNA2 with the best inhibition of gene inhibition lay the foundation for further experiments.