摘要
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。
Abstract This study was aimed to explore the effect of bortezomib on proliferation and apoptosis of acute monocytic leukemic cells SHI-1 and the function of Bcl-2 gene family including Bc12112, Bcl-2 and Bax in its apoptosis. SHI-1 cells were cultured and treated with bonezomib of different concentrations for different time. MTT assay was used to de- tect the proliferation and apoptosis, Annexin-V staining, mitochondrial transmembrane potential （A0m） and DNA aga- rose gel eleetrophoresis were used to investigate apoptosis of SHI-1 cells. RT-PCR was used to analyze the levels of Bc12112, Bcl-2 and bax mRNA in SHI-1 cells treated with bortezomib for 0,6,12 and 24 hours. The results showed that bortezomib inhibited the proliferation of SHI-1 ceils in time-and doze-dependent manners, the IC50 at 24 and 48 hours were 54.13 nmol/L and 5.45 nmol/L respectively. Bortezomib could induce apoptosis of SHI-1 cells in time-dependent manner, increase expression of Annexin-V positive cells, deerease A0m of SHI-1 cells and result in DNA fragmention and morphologic changes of apoptosis. RT-PCR showed that Bc12112 mRNA expression was up-regulated, bcl-2 mRNA expression was down-regulated and bax mRNA expression was not changed obviously. It is concluded that bortezomib inhibits the proliferation of SHI-1 and induces apoptosis in which Bc12112 and Bcl-2 gene can be ones of the main genes taking part in.
出处
《中国实验血液学杂志》
CAS
CSCD
2008年第5期1016-1020,共5页
Journal of Experimental Hematology
基金
宁波市医学科技计划项目资助(编号200520)
宁波市干细胞移植重点实验室资金资助(编号2006A62003)
