摘要
目的研究α1,4半乳糖糖基转移酶(α1,4GalT)基因反义寡核苷酸(antisenseoligonucleotide,ASO)对脑胶质瘤细胞系SWO-38细胞的增值与凋亡的影响和与Fas表达的关系。方法采用脂质体介导的基因转染方法将α1,4GalT基因ASO转入SWO-38细胞中。应用RT-PCR检测对α1,4GalTmRNA表达的影响,克隆形成试验检测对细胞增值的作用,流式细胞术(FCM)和琼脂糖凝胶电泳检测细胞凋亡的发生,FACS检测凋亡相关蛋白Fas表达的变化。结果脂质体介导的ASO转染后,经RT-PCR检测证实α1,4GalTmRNA基因的表达下降;克隆形成试验显示细胞生长明显受到抑制(P<0.01);DNA电泳图像呈凋亡表现;FCM检测表明,导入α1,4GalT基因ASO的SWO-38细胞的凋亡明显增加(P<0.01);Fas蛋白表达水平明显下降(P<0.01)。结论α1,4GalT基因ASO可诱导脑胶质瘤细胞系SWO-38细胞凋亡发生,其机制可能与fas基因调控有关。
Objective To study the effects of α1,4Gal T antisense oligonucleotide mediated by lipofectin on human glioma cell line SWO-38. Methods SWO38 glioma cells were exposed to 10 μmol/L α1,4Gal T antisense oligonucleotide for 72 h by means of lipofectin transfection, and the growth inhibition of the cells was detected by colony-forming unit assay. Analysis of DNA fragmentation was performed with flow cytometry (FCM) and agarose gel eletrophoresis with Fas protein expression determined by FCM. Resultsα1,4Gal T antisense oligonucleotide significantly inhibited the growth of glioma cells (P<0.01) and induced apoptosis in SWO-38 cells (P<0.05). Marked up-regulation of Fas protein expression was observed in response to the treatment (P<0.01). Conclusionα1,4Gal T antisense oligonucleotide can significantly inhibit SWO-38 cell proliferation and induce cellular apoptosis, the mechanism of which may involve up-regulated Fas expression.
出处
《第一军医大学学报》
CSCD
北大核心
2002年第4期299-302,共4页
Journal of First Military Medical University
基金
国家自然科学基金(397100143)
国家自然科学基金(39870813)
