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本研究探讨蛋白酶抑制剂硼替佐米对急性单核细胞白血病细胞株SHI-1增殖和凋亡的影响及Bcl2l12、Bcl-2和Bax基因在其中的作用。用MTT比色法观察硼替佐米对SHI-1细胞的生长抑制作用,用Annexin-V标记、线粒体跨膜电位(Δψm)和DNA凝胶电泳分析细胞凋亡,用RT-PCR方法检测0、6,12和24小时Bcl2l12、Bcl-2和Bax基因表达。结果表明,硼替佐米呈时间和剂量依赖性抑制SHI-1细胞生长,24小时和48小时半数抑制浓度分别为54.13 nmol/L和5.45 nmol/L;硼替佐米能够诱导SHI-1细胞凋亡,在6小时Annexin-V阳性细胞就开始增高并呈时间依赖性,Δψm减低,形态学可见明显细胞核凝聚、固缩和碎裂,DNA凝胶电泳显示DNA片段化;RT-PCR显示,Bcl2l12表达增高,Bcl-2表达减低,但Bax表达无明显改变。结论:硼替佐米抑制SHI-1细胞增殖,并可能通过上调Bcl2l12基因和下调Bcl-2基因,使线粒体膜电位下降而促使SHI-1细胞发生凋亡。
This study was aimed to investigate the effect of bortezomib, a protease inhibitor, on the proliferation and apoptosis of acute monocytic leukemia cell line SHI-1 and the role of Bcl-2, Bcl-2 and Bax genes in this study. The inhibitory effect of bortezomib on the growth of SHI-1 cells was observed by MTT colorimetric assay. The apoptosis of SHI-1 cells was detected by Annexin-V, mitochondrial transmembrane potential (Δψm) and DNA gel electrophoresis. Bcl2112, Bcl-2 and Bax gene expression at 6, 12 and 24 hours. The results showed that bortezomib could inhibit the growth of SHI-1 cells in a dose-dependent and time-dependent manner. The inhibitory concentrations of 54.1 and 54.5 nmol / L were 24.13 and 5.45 nmol / L, respectively. Bortezomib could induce SHI-1 cell apoptosis , Annexin-V positive cells began to increase in a time-dependent and time-dependent manner at 6 hours, Δψm decreased, the morphology showed obvious nuclear condensation, condensation and fragmentation, DNA gel electrophoresis showed DNA fragmentation; RT-PCR showed that Bcl2l12 expression Increased, Bcl-2 expression decreased, but no significant change in Bax expression. CONCLUSION: Bortezomib can inhibit the proliferation of SHI-1 cells and induce the apoptosis of SHI-1 cells by decreasing the mitochondrial membrane potential by up-regulating Bcl-212 gene and down-regulating Bcl-2 gene.