Mu Qi-Tian, Ouyang Gui-Fang, Lou Yan-Ru, Chen Xiao-Pei, Lu Ying, Liang Wei, Zhang Yi, Xu Wei
Ningbo First Hospital, Ningbo Laboratory of Stem Cell Transplantation, Ningbo 315010, Zhejiang Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Oct;16(5):1016-20.
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 Bcl2l12, Bcl-2 and Bax in its apoptosis. SHI-1 cells were cultured and treated with bortezomib of different concentrations for different time. MTT assay was used to detect the proliferation and apoptosis, Annexin-V staining, mitochondrial transmembrane potential (DeltaPsim) and DNA aga-rose gel electrophoresis were used to investigate apoptosis of SHI-1 cells. RT-PCR was used to analyze the levels of Bcl2l12, 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 cells in time-and doze-dependent manners, the IC(50) 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, decrease DeltaPsim of SHI-1 cells and result in DNA fragmentation and morphologic changes of apoptosis. RT-PCR showed that Bcl2l12 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 Bcl2l12 and Bcl-2 gene can be ones of the main genes taking part in.
本研究旨在探讨硼替佐米对急性单核细胞白血病细胞SHI-1增殖和凋亡的影响，以及包括Bcl2l12、Bcl-2和Bax在内的Bcl-2基因家族在其凋亡过程中的作用。培养SHI-1细胞，并用不同浓度的硼替佐米处理不同时间。采用MTT法检测细胞增殖和凋亡情况，通过膜联蛋白V染色、线粒体跨膜电位（ΔΨm）检测及DNA琼脂糖凝胶电泳研究SHI-1细胞凋亡情况。采用RT-PCR分析硼替佐米处理0、6、12和24小时的SHI-1细胞中Bcl2l12、Bcl-2和bax mRNA的水平。结果显示，硼替佐米能以时间和剂量依赖性方式抑制SHI-1细胞增殖，24小时和48小时的半数抑制浓度（IC50）分别为54.13 nmol/L和5.45 nmol/L。硼替佐米能以时间依赖性方式诱导SHI-1细胞凋亡，增加膜联蛋白V阳性细胞的表达，降低SHI-1细胞的ΔΨm，并导致DNA片段化和凋亡形态学改变。RT-PCR结果显示，Bcl2l12 mRNA表达上调，bcl-2 mRNA表达下调，bax mRNA表达无明显变化。结论：硼替佐米抑制SHI-1细胞增殖并诱导其凋亡，其中Bcl2l12和Bcl-2基因可能是参与其中的主要基因。
