Livin反义寡核苷酸对骨肉瘤MG-63细胞活性的影响

目的 探讨Livin在骨肉瘤细胞中的表达及Livin反义寡核苷酸（ASODN）对骨肉瘤细胞活性及功能的影响。方法 设计针对Livin的ASODN序列，根据转染效率评价结果，选择下调作用最强的ASODN序列制备脂质体-寡核苷酸复合物并转染 MG-63 细胞。采用免疫细胞染色法及Western blotting检测转染24 h后MG-63 细胞中Livin的表达情况，CellTiter-Glo法检测转染72 h后MG-63 细胞的增殖情况，划痕法检测ASODN对MG-63细胞迁移能力的影响，流式细胞仪检测转染24 h后 MG-63 细胞的凋亡情况和细胞周期。结果 Livin蛋白主要表达于MG-63细胞的胞质。选用下调作用最强的ASODN转染MG-63细胞后，Livin蛋白表达显著下降；不同浓度Livin ASODN 对 MG-63细胞的增殖及迁移有剂量依赖抑制作用；而ASODN 组的凋亡率明显高于空白对照组和阴性对照组（P＜0.05）。结论 靶向Livin的ASODN可以显著抑制骨肉瘤细胞增殖和侵袭能力并促进其凋亡。

Impact of Livin antisense oligonucleotide on the activity of osteosarcoma MG-63 cells

Objective To investigate the expression of Livin in osteosarcoma MG-63 cells and the effect of Livin anti-sense oligonucleotide(ASODN) on its activity and function.Methods The ASODN targeting Livin were designed and constructed.According to the results of transfection efficiency evaluation, the liposome/oligonucleotide complexes were prepared by adding oligonucleotide and then transferred into MG-63 cells.The immunohistochemical staining and Western blotting were used to detect the expression of Livin at 24th h after transfection in MG-63 cells.The proliferation at 72nd h and apoptosis at 24th h after transfection were measured by CellTiter-Glo and Flow cytometry,respectively.Results The Livin was mainly expressed in the cytoplasm of MG-63 cells.We chose ASODN with the highest transfection efficiency to transfer MG-63 cells.The expression of Livin protein decreased significantly after the transfection, and Livin ASODN at different concentrations inhibit the proliferation and migration of MG-63 cells with significant difference.The apoptotic rate of Lip-ASODN group was higher than those of blank control group and negative control group in a dose-dependent manner (P<0.05).Conclusion The ASODN targeting Livin can inhibit the proliferative capacity and promote the apoptosis of osteosarcoma MG-63 cells.