生存素反义寡核苷酸诱导脑胶质瘤C6细胞凋亡的实验研究

Researches of Survivin antisense oligonucleotides inducing the apoptosis of neurogliocytoma C6 cells

目的观察生存素(Survivin)反义寡核苷酸(ASODN)抑制Survivin蛋白表达对胶质瘤C6细胞凋亡、增殖的影响。方法用载有Survivin mRNA特异性ASODN的脂质体转染C6细胞后,用Western blot法检测Survivin蛋白表达情况,MTT实验观察C6细胞生长抑制情况,流式细胞术分析细胞凋亡程度。结果脂质体及Survivin正义寡核苷酸(SODN)对C6细胞生长、Survivin蛋白表达无明显影响(P>0.05),而Survivin A-SODN对C6细胞生长及Survivin蛋白表达有明显的抑制作用(P<0.01);以脂质体单独转染后,C6细胞凋亡水平无明显变化(P>0.05),以SODN转染后则较多细胞发生了凋亡(与空白对照组比较,P<0.05),而SurvivinASODN转染后C6细胞凋亡数显著增多(与其他各组比较,P<0.01)。结论Survivin ASODN能靶向抑制Survivin蛋白的表达,从而解除其对凋亡的抑制作用,诱导胶质瘤细胞凋亡的发生,最终抑制胶质瘤细胞的生长。

[Objectives] To observe the effects of inhibition of Survivin protein expression by Survivin antisense oligonucleotides （ASODN） on the apoptosis and proliferation of neuroglioeytoma C6 cells. [Methods] Measuring the Survivin protein expression by Western blot after transfecting the C6 cells with liposomes carrying Survivin mRNA specific ASODN, and the growth inhibition of C6 cells was observed by MTT experiment and the apoptosis degree was analyzed by flow cytometry. [Results] Liposomes and Survivin sense ohgonucleotide （SODN） had no significant effects on the growth of C6 cells and the expression of Survivin protein （P 〉0.05）, but Survivin ASODN showed significant inhibition on the growth of C6 cells and the expression of Survivin protein （P 〈0.01）; the apoptosis level of C6 cells did not show significant changes after transfecting with liposome alone （P 〉0.05）, and relatively more cells underwent apoptosis after they were transfected with SODN （P 〈0.05, in comparison with the blank control group）, whereas the apoptotic number of C6 cells significantly increased after they were transfected with Survivin ASODN （P 〈0.01, in comparison with other groups）. [Conclusion] Survivin ASODN can directly inhibit the expression of Survivin protein, and subsequently eliminate the inhibitory effects on apoptosis, induce the occurrence of neurogliocytoma apoptosis and finally inhibit the growth of neurogliocytoma cells.