陷阱受体3反义寡核苷酸对胶质瘤U251细胞凋亡的影响

Effect of DcR3 antisense oligonucleotides on apoptosis of glioma cells

目的观察陷阱受体3（DcR3）反义核苷酸（ASODN）在胶质瘤细胞凋亡中的作用。方法构建并挑选DcR3反义核苷酸，转染U251细胞，逆转录-聚合酶链反应（RT—PCR）、Western blot方法观察DcR3 mRNA和蛋白有无降低，流式细胞仪观察细胞凋亡的变化。结果经1．2μmol／L AS20DN处理的U251细胞可有效地降低DcR3 mRNA和蛋白表达，DcR3／β-actin比值分别为（12．4±3．1）％和（13．5±4．2）％，错义链组、脂质体组、对照组mRNA和蛋白分别为（68．8±4．5）％、（74．5±5．0）％、（71．3±6．4）％和（78．2±6．8）％、（79．8±4．5）％、（76．4±5．6）％，差异有统计学意义（P〈0．05），同时，传染DcR3反义核苷酸的U251细胞凋亡率为（42．9±13．1）％，明显高于错义链组（15．8±4．8）％、脂质体组（15．7±3．7）％和对照组（14．6±4．0）％，差异有统计学意义（P〈0．05）。结论DcR3反义核苷酸可使U251细胞凋亡增多。

Objective To explore the effect of DcR3 antisense oligonucleotides on apoptosis of glioma ceils. Methods After constructing and choosing DcR3 antisense oligonucleotides and transfecting glioma cell lines U251 ,we detected the DcR3 mRNA and protein expression by RT-PCR and Western blot assays,and observed the apoptosis by flow cytometry. Results 1.2 μmol/L DcR3 antisense oligonucleotides could effectively reduce the expression of DcR3 mRNA and protein （ P 〈 0.05 ） with the ratio of DcR3/β-actin being （ 12.4 ± 3. 1 ） % and （ 13.5 ± 4.2 ） % respectively. In the SC, LFA, and control groups,the expression of DeR3 mRNA and protein was （68.8 ± 4.5 ）%, （74.5 ±5. 0）%, （71.3± 6.4 ） %, and （78.2 ± 6.8 ） %, （ 79.8± 4.5 ） %, （ 76.4± 5.6 ） % respectively. The apoptosis rate in glioma cells transfected with DcR3 antisense oligonucleotides was （42.9 ± 13.1 ）% ,significantly higher than （ 15.8 ± 4.8 ） %, （ 15.7 ± 3.7） %, （ 14.6± 4.0） % respectively in the SC, LFA, and control groups （ P 〈 0.05）. Conclusion DcR3 antisense oligonucleotides could induce apoptosis of glioma cells.