摘要
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,Cell TiterGlo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24h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。
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 24^t h after transfection in MG-63 cells.The proliferation at 72^nd h and apoptosis at 24^th 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 transfeetion efficiency to transfer MG-63 cells. The expression of Livin protein decreased significantly after the transfec- tion, 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.
出处
《临床肿瘤学杂志》
CAS
2017年第6期493-499,共7页
Chinese Clinical Oncology
基金
江苏省自然科学基金面上项目(BK20151373)
