全硫代反义寡核苷酸对乳腺癌MDA-MB-231细胞的影响

摘要

目的 本研究探讨全硫代反义寡核苷酸( PS-ASODN)对乳腺癌MDA-MB-231细胞端粒酶活性及细胞生长、迁移、侵袭的影响.方法 本实验将PS-ASODN经脂质体转染作用于乳腺癌MDA-MB-231细胞,将实验分为空白对照组、对照正义全硫代寡核苷酸(PS-SODN)组和不同剂量的PS-ASODN组；脂质体介导的PS-ASODN和PS-SODN作用于乳腺癌MDA-MB-231细胞后,分别采用四甲基偶氮唑蓝比色法(MTT)、酶联免疫吸附法(EUSA)、流式细胞术、Transwell小室迁移、侵袭实验检测细胞的体外增殖、端粒酶活性、细胞凋亡、细胞迁移、侵袭能力的影响.结果 终浓度为1、3及5 μmol/L的PS-ASODN对MDA-MB-231细胞端粒酶活性及细胞的增殖均有抑制作用,与空白对照组比较差异显著(P＜0.01),并呈一定剂量依赖性；ELISA法检测结果,1、3及5μmol/L的PS-ASODN对MDA-MB-231细胞作用72 h后端粒酶皆为阴性,表明端粒酶PS-ASODN能够抑制端粒酶活性,对照组端粒酶皆为阳性.1、3及5μmol/L的PS-ASODN作用24h可以明显抑制MDA-MB-231细胞的迁移和侵袭能力(P＜0.01).结论 PS-ASODN能有效抑制人乳腺癌MDA-MB -231细胞的生长、促进凋亡、抑制其侵袭和迁移能力,其机制可能是通过PS-ASODN降低端粒酶活性而实现.%Objective To study the effects of phosphorothioate sense antisense oligodeoxy nucleotides (PS-ASODN) on telomerase activity, apoptosis, migration and invasion of MDA-MB-231 cell line. Methods MDA-MB-231 cells were treated with PS-ASODN liposome. The anti-proliferation effects on MDA-MB-231 cells were determined by MTT assay. The activity of telomerase was tested by ELSIA. Apoptosis and cell cycle were examined by flow cytometer method. Transwell invasive was used to observe the change of the invasion and migration abilities. Results 1, 3 and 5 μmol/L PS-ASODN significantly inhibited the growth of MDA-MB-231 cell line and the inhibition rates were higher than those in control group (P <0.01). Telomerase of MDA-MB-231 cell was repressed by 1, 3 and 5 μmol/L telomerase PS-ASODN after 72 hours later, it showed that telomerase PS-ASODN could inhibit the activity of telomerase. After treated by 1, 3 and 5 μmol/L PS-ASODN for 24 hours, the invasive and migratory abilities were inhibited conspicuously (P<0. 01). Conclusion PS-ASODN targeted human telomerase not only inhibits MDA-MB-231 cell proliferation and telomerase activity, but also inhibits the invasive and migratory abilitiers of MDA-MB-231 cell line.