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目的探讨通过siRNA沉默细胞TAp73蛋白表达对骨肉瘤Saos-2细胞的增殖、周期以及侵袭力的作用研究。方法以骨肉瘤Saos-2细胞系为研究对象,设计合成沉默TAp73的siRNA片段,转染入Saos-2细胞,用Western-blot法检测干扰后TAp73蛋白的表达水平,CCK-8法检测细胞增殖,利用流式细胞仪检测细胞周期,Transwell法检测细胞侵袭能力。结果 Western-blot检测显示siRNA对TAp73蛋白表达有显著下调作用;CCK-8检测实验组细胞增殖明显(P<0.05);细胞周期检测显示细胞处于DNA合成期(S期)的细胞比例上升;穿过人工基底膜的细胞数量也明显增加,由(84.17±1.35)增加为(92.67±2.80)(P<0.05)。结论抗癌基因TAp73在骨肉瘤的增殖、凋亡和侵袭过程中发挥有重要作用,可能成为控制肿瘤增殖和转移的关键性因素。
Objective To investigate the effect of silencing the expression of TAp73 by siRNA on the proliferation, cycle and invasiveness of osteosarcoma Saos-2 cells. Methods The siRNA targeting TAp73 was designed and synthesized in Saos-2 cell line of osteosarcoma and transfected into Saos-2 cells. The expression of TAp73 protein was detected by Western-blot, and the cell proliferation was detected by CCK-8 , The cell cycle was detected by flow cytometry, and the invasion ability was detected by Transwell method. Results The results of Western-blot showed that siRNA significantly down-regulated the expression of TAp73 protein. The cell proliferation was significantly increased in CCK-8 assay group (P <0.05). The cell cycle assay showed that the proportion of cells in the DNA synthesis phase (S phase) The number of cells in the artificial basement membrane also significantly increased from (84.17 ± 1.35) to (92.67 ± 2.80) (P <0.05). Conclusion The anti-oncogene TAp73 plays an important role in the proliferation, apoptosis and invasion of osteosarcoma and may play a key role in the control of tumor proliferation and metastasis.