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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 investigate the effects of α1,4GalT gene antisense oligonucleotide (ASO) on the proliferation and apoptosis of glioma cell line SWO-38 and its relationship with the expression of Fas Relationship. Methods The α1,4GalT gene ASO was transfected into SWO-38 cells by liposome-mediated gene transfection. The effect of α1,4GalTmRNA expression on α1,4GalTmRNA was detected by RT-PCR. The effect of cell proliferation was detected by clone formation assay. The apoptosis was detected by flow cytometry (FCM) and agarose gel electrophoresis. FACS was used to detect the expression of apoptosis related protein Fas Changes in expression. Results After ASO transfection, the expression of α1,4GalTmRNA was down-regulated by RT-PCR. The clonal formation assay showed that the cell growth was significantly inhibited (P <0.01); the DNA electrophoresis image showed apoptosis; FCM The results showed that the apoptosis of SWO-38 cells transfected with α1,4GalT gene ASO was significantly increased (P <0.01), and the expression of Fas protein was significantly decreased (P <0.01). Conclusion α1,4GalT gene ASO can induce the apoptosis of glioma cell line SWO-38, which may be related to the regulation of fas gene.