Chk1反义寡核苷酸影响胶质瘤放疗敏感性研究.docVIP

Chk1反义寡核苷酸影响胶质瘤放疗敏感性研究 　　 【摘要】 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Western Blot法检测Chk1蛋白，Real time PCR检测Chk1 mRNA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。 　　 【关键词】 胶质瘤； 细胞周期检测点激酶； 反义寡核苷酸； 放射敏感性 　　 　　 Transfection of Chk1 Antisense Oligonucleotide to Glioma Increases the Apoptotic Sensitivity to Irradiation/LI Yong， LAI Run-long， TAN Dian-hui，et al.//Medical Innovation of China，2012，9(13)：003-005 　　 【Abstract】 Objective：Effect on expression of Chk1 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 Chk1 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 Chk1 sense and antisense chain. The expression of Chk1 was measured by Western blot， and Chk1 mRNA was measured by real time PCR.Results：The expression of Chk1 protein and mRNA markedly decreased after ransfecting U251 cell line with Chk1 antisense oligonucleotide.Transfecting U251 cell line with Chk1 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 Chk1 antisense oligonucleotide to glioma. This can be the theoretical basis for increasing the apoptotic sensitivity to irradiation of glioma. 　　 【Key words】 Glioma; Checkpoint kinase; Antisense oligonucleotide; Irradiation 　　 First-author’s address：First Affiliated Hospital of Shantou University Medical College，Shantou 515041，China 　　 doi：10.3969/j.issn.1674-4985.2012.13.002 　　 　　 目前研究认为，细胞周期检测点信号传导通路主要由ATM/ATR-Chk1/2-Cdc25A、Cdc25B、Cdc25c-Cdk轴组成[