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目的:通过抑制人RAD51修复基因(hRAD51)表达,研究hRAD51对骨肉瘤细胞和裸鼠移植瘤生长的影响。方法:构建hRAD51基因沉默的骨肉瘤细胞株HOS-shA2,利用MTT和克隆形成实验从体外观察hRAD51基因沉默对HOS细胞增殖的影响,FCM法检测抑制hRAD51基因表达后细胞周期的变化情况,裸鼠成瘤实验观察降低hRAD51表达对骨肉瘤移植瘤生长情况的影响。结果:成功构建了hRAD51基因沉默细胞株HOS-shA2;体外实验显示,HOS-shA2细胞的增殖能力较亲本及阴性对照细胞明显降低(P<0.01);FCM法检测结果显示,hRAD51基因沉默后G2/M期细胞比例增多(P<0.01);HOS-shA2细胞的裸鼠移植瘤生长速度缓慢,剥离瘤体质量明显轻于亲本HOS组和阴性对照组,差异有统计学意义(P<0.01)。结论:hRAD51在骨肉瘤细胞增殖中发挥重要作用,有望成为肿瘤治疗的新靶点。
OBJECTIVE: To investigate the effect of hRAD51 on the growth of human osteosarcoma cells and xenografts in nude mice by inhibiting the expression of human RAD51 repair gene (hRAD51). METHODS: Human osteosarcoma cell line HOS-shA2 silenced by hRAD51 gene was constructed. The effect of hRAD51 gene silencing on the proliferation of HOS cells was observed in vitro using MTT and colony formation assay. Changes of cell cycle after hRAD51 gene expression inhibition by FCM were detected. The effect of hRAD51 expression on the growth of transplanted tumors of osteosarcoma was observed by tumor formation. RESULTS: The hRAD51 gene silencing cell line HOS-shA2 was successfully constructed. In vitro experiments showed that the proliferation of HOS-shA2 cells was significantly lower than that of parental and negative control cells (P<0.01). The results of FCM showed that hRAD51 gene was silenced after G2 The proportion of cells in /M phase increased (P<0.01); the growth rate of transplanted tumors of HOS-shA2 cells in nude mice was slow, and the quality of stripped tumors was significantly lighter than that of parental HOS and negative controls (P<0.01). . Conclusion: hRAD51 plays an important role in the proliferation of osteosarcoma cells and is expected to become a new target for cancer therapy.