Survivin反义寡核苷酸抑制EC9706细胞增殖并诱导细胞凋亡

Induction of Apoptosis and Inhibition of EC9706 Cell Proliferation by Survivin Antisense Oligonucleotide

目的观察Survivin反义寡核苷酸(ASODN)对人食管癌细胞系EC9706细胞增殖和凋亡的影响。方法人工合成Survivin基因反义和正义ODN,并进行硫代磷酸化修饰,通过脂质体途径分别转染EC9706细胞;应用RT-PCR和Western Blot检测Survivin mRNA和蛋白表达;应用MTT法检测Survivin ASODN对EC9706细胞增殖的影响;流式细胞仪检测细胞周期变化及细胞凋亡比率。结果体外培养的EC9706细胞可表达较强的Survivin mRNA和蛋白;Survivin ASODN可呈浓度依赖性地抑制Survivin mRNA和蛋白表达,50μmol/LASODN几乎可以完全抑制。MTT研究结果表明,Survivin ASODN可呈浓度依赖性地抑制EC9706细胞增殖,50μmol/LASODN对细胞生长的抑制率可达78.5%,诱导细胞凋亡,使细胞阻滞于G2/M期。Survivin SODN对Survivin mRNA和蛋白以及EC9706细胞的增殖和细胞周期无明显的抑制作用。结论脂质体介导转染Survivin反义寡核苷酸可抑制细胞增殖、诱导细胞G2/M期阻滞而促进细胞凋亡。

Objective To observe the effects of Survivn antisense oligonucleotide （ASODN） on cell proliferation and apoptosis of EC9706 cells. Methods Synthetic ODN was completely phosphorothioate-modified. ASODN was transfected into EC9706 cells. The expressions of Survivin mRNA and protein were detected by RT-PCR and Western Blot. MTT was used as the method of EC9706 proliferation observation. The cell-cycle and apoptosis were analyzed by flow cytometry. Results EC9706 cells spontaneously expresseds Survivin mRNA and protein. Both mRNA and protein expressions of Surviving decreased significantly in the ASODN transfected cells comparing with those in the blank control cells and cells stransfected with SODN. Survivin ASODN significantly inhibited cell proliferation and induced apoptosis in a dose-dependent manner. The cell-cycle in the ASODN transfected cells was arrested at the G2/M phase. Conclusion ASODN targeting at Survivin mRNA can inhibit EC9706 cell proliferation and induce G2/M arrest and apoptosis.