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目的利用RNAi抑制Rad51蛋白表达来研究Rad51在骨肉瘤治疗中的作用。方法western blot和RT-PCR观察Rad51在骨肉瘤细胞中蛋白和RNA水平的变化。采用RNA干扰技术,构建shRNA-Rad51载体。用western blot和RT-PCR方法证实RNA干扰的有效性和可行性。用MTT和克隆形成实验研究RNAi抑制Rad51表达对γ-射线的敏感性。流式分析法观察RNAi抑制Rad51表达对细胞凋亡的影响。结果成功构建了shRNA-Rad51载体。RNAi抑制Rad51表达增强骨肉瘤细胞对γ-射线的敏感性。RNAi抑制Rad51表达促进骨肉瘤细胞的凋亡。结论RNAi抑制Rad51表达在骨肉瘤治疗中有作用,有望成为骨肉瘤治疗的新靶点。
Objective To study the role of Rad51 in the treatment of osteosarcoma using RNAi to inhibit Rad51 protein expression. Methods Western blot and RT-PCR were used to detect the expression of Rad51 in osteosarcoma cells. RNA interference technology was used to construct shRNA-Rad51 vector. The effectiveness and feasibility of RNA interference were confirmed by western blot and RT-PCR. The sensitivity of RNAi to γ-rays inhibition of Rad51 expression was investigated by MTT and clonogenic assays. Flow cytometry was used to observe the effect of RNAi on Rad51 expression on apoptosis. Results shRNA-Rad51 vector was successfully constructed. Inhibition of Rad51 expression by RNAi enhances the sensitivity of osteosarcoma cells to γ-rays. RNAi Inhibits Rad51 Expression and Promotes Apoptosis of Osteosarcoma Cells. Conclusion RNAi can inhibit Rad51 expression in the treatment of osteosarcoma and is expected to become a new target for the treatment of osteosarcoma.