论文部分内容阅读
目的运用RNAi慢病毒沉默CD59的表达,观察其对急性T淋巴细胞白血病Jurkat细胞株增殖、凋亡的影响。方法构建CD59 RNAi-EGFP融合蛋白慢病毒载体,转染急性T系白血病Jurkat细胞株,筛选出稳定转染的细胞系,空病毒组为阴性对照,未处理的Jurkat细胞系作为空白对照;荧光显微镜和流式细胞仪观察各组的细胞转染效率;ELISA检测各组细胞CD59的表达情况;RT-PCR检测各组CD59基因以及凋亡相关基因Bcl-2/Bax m RNA的表达;CCK8表达检测细胞增殖效率的改变;流式细胞术检测各组细胞凋亡情况。结果荧光显微镜和FCM观察转染效率在90%以上;ELISA结果显示实验组细胞CD59蛋白表达降低;RT-PCR结果显示实验组CD59、Bcl-2 m RNA表达水平降低(P<0.05),Bax m RNA表达水平升高(P<0.05);CCK8结果显示实验组细胞增殖效率明显降低(P<0.05);流式细胞仪结果显示沉默CD59的表达能够促进细胞凋亡(P<0.05)。结论沉默D59基因表达可抑制急性T系白血病Jurkat细胞株的增殖能力并诱导细胞凋亡,为临床急性T系白血病的治疗提供了新靶标、新思路。
Objective To silence the expression of CD59 by RNAi lentivirus and observe its effect on the proliferation and apoptosis of Jurkat cell line of acute lymphoblastic leukemia. Methods CD59 RNAi-EGFP fusion lentiviral vector was constructed and transfected into Jurkat cell line of acute T-leukemia. Stably transfected cell lines were screened out. Negative control was in the empty virus group and untreated Jurkat cell line was used as a control. Fluorescence microscopy The expression of CD59 and Bcl-2 / Bax m RNA in each group were detected by RT-PCR. The expression of CCK8 Cell proliferation efficiency changes; flow cytometry detection of apoptosis in each group. Results The transfection efficiency was above 90% by fluorescence microscopy and FCM. The results of ELISA showed that the expression of CD59 protein in the experimental group was decreased. The expression of CD59 and Bcl-2 mRNA in the experimental group was decreased (P <0.05) (P <0.05). The results of CCK8 showed that the cell proliferation efficiency was significantly decreased in experimental group (P <0.05). The results of flow cytometry showed that silencing CD59 expression could promote apoptosis (P <0.05). Conclusion Silencing D59 gene expression can inhibit the proliferation of Jurkat cells and induce apoptosis in acute T-leukemia cells. It provides new targets and new ideas for the treatment of acute T-cell leukemia.