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本研究探讨新的白血病相关基因zo-1在白血病细胞中的作用及其初步机制。应用甲基化PCR方法验证THP-1等白血病细胞中基因zo-1甲基化与白血病的关系,用甲基化PCR及RT-PCR方法验证THP-1等白血病细胞系中基因zo-1甲基化与基因表达的关系。选择基因zo-1高表达的THP-1细胞,用脂质体Oligofectamine将针对基因zo-1的小干扰RNA转染THP-1细胞,RT-PCR方法验证抑制效率,AnnexinV和PI观察细胞凋亡率,PI检测细胞周期,CCK-8检测细胞增殖。结果表明:急性白血病人中基因zo-1呈高甲基化状态,健康人群中基因zo-1不发生甲基化。基因zo-1未甲基化的THP-1细胞系高表达基因zo-1;基因zo-1高甲基化的Molt4及HL-60细胞系中不表达基因zo-1。脂质体Oligofectamine转染针对zo-1的小干扰RNA成功地抑制了THP-1细胞基因zo-1的表达,而不影响THP-1细胞增殖、细胞周期及细胞凋亡。结论:基因zo-1为白血病相关基因,急性白血病中基因zo-1高甲基化,基因zo-1的甲基化状态调控基因zo-1表达。THP-1细胞中基因zo-1表达缺失不影响细胞增殖、细胞周期及细胞凋亡。
This study was to investigate the role of novel leukemia-associated gene zo-1 in leukemia cells and its primary mechanism. The methylation of zo-1 methylation in leukemia cells such as THP-1 and leukemia was verified by methylation PCR. The gene zo-1 in leukemia cell line THP-1 was verified by methylation PCR and RT-PCR Relationship between gene expression and gene expression. THP-1 cells with high expression of zo-1 gene were selected and transfected with small interfering RNA targeting zo-1 gene into THP-1 cells by Lipofectamine. The inhibitory rate was determined by RT-PCR, apoptosis was observed by Annexin V and PI Rate, PI detection of cell cycle, CCK-8 detection of cell proliferation. The results showed that: in patients with acute leukemia gene zo-1 hypermethylation state, healthy people in the gene zo-1 does not occur methylation. Gene zo-1 unmethylated THP-1 cell line overexpression gene zo-1; gene zo-1 hypermethylation Molt4 and HL-60 cell line does not express the gene zo-1. Small interfering RNA transfected with liposome Oligofectamine against zo-1 successfully inhibited the expression of zo-1 in THP-1 cells without affecting the proliferation, cell cycle and apoptosis of THP-1 cells. Conclusion: The gene zo-1 is a leukemia-related gene. The gene zo-1 hypermethylation and the zo-1 methylation status zo-1 gene expression in acute leukemia. Loss of zo-1 gene expression in THP-1 cells did not affect cell proliferation, cell cycle and apoptosis.