Chk1反义寡核苷酸影响胶质瘤放疗敏感性的研究

Transfection of Chk1 Antisense Oligonucleotide to Glioma Increases the Apoptotic Sensitivity to Irradiation

目的:观察转染Chk1反义寡核苷酸(ASON)后,对照射后U251细胞株中Chk1表达、细胞周期及细胞凋亡的影响。方法:采用脂质体转染法,Chkl的正义、反义寡核苷酸对U251细胞株进行Chkl转染。以放射线照射后,测定其细胞周期和凋亡率变化,比较Chkl转染正义链和反义链对细胞放射敏感性的不同。用WesternBlot法检测Chk1蛋白,RealtimePCR检测Chk1mRNA表达。结果:转染Chk1反义寡核苷酸后,能明显下调Chk1蛋白和mRNA的表达,显著增强放射线诱导的肿瘤细胞凋亡,并消除细胞周期阻滞。结论:反义核酸技术灭活Chk1基因显著增强放疗诱导的U251细胞凋亡,为增敏胶质瘤的放射治疗提供了理论依据。

Objective ：Effect on expression of Chkl and changes of cell cycle after radiation in U251 cell line with antisense oligodeoxynucleotide （ASON） were observed.Methods ：The U251 cell line was transfected with Chkl sense and antisense chain-lipofectamine PLUS complex. Then irradiated it and measured the changes of the cell cycle and apoptosis in order to compare the diference between the Chkl sense and antisense chain. The expression of Chkl was measured by Western blot, and Chkl mRNA was measured by real time PCR.Results ：The expression of Chkl protein and mRNA markedly decreased after ransfecting U251 cell line with Chkl antisense oligonucleotide.Transfecting U251 cell line with Chkl antisense oligonucleotide could increase apoptosis significantly and decrease cell cycle arrest markedly induced by irradiation.Conclusion ：The sensitivity to apoptosis is excessively increased after treated with irradiation by transfection of Chkl antisense oligonucleotide to glioma. This can be the theoretical basis for increasing the apoptotic sensitivity to irradiation of glioma.