摘要
为了了解三尖杉酯碱（ＨＴ）对白血病癌细胞杀伤的分子机制，用间接免疫荧光、免疫印迹、分子杂交和点杂交的方法探讨了ＨＴ的抗急性粒细胞白血病ＨＬ－６０效应和动粒／着丝粒蛋白之间的关系，结果显示：（１）抗动粒抗体（ａｎｔｉｃｅｎｔｒｏｍｅｒｅａｎｔｉｂｏｄｙ，ＡＣＡ）血清的间接免疫荧光染色法显示，ＨＬ６０细胞中动粒荧光斑点随ＨＴ作用时间延长而减弱，表明细胞内动粒蛋白含量降低。（２）Ｗｅｓｔｅｒｎｂｌｏｔ法显示，ＡＣＡ血清能识别ＨＬ６０细胞中６种分子量大小不同的动粒蛋白：１４０ｋＤ、８０ｋＤ、７０ｋＤ、５６ｋＤ、５２ｋＤ和１７ｋＤ。ＨＴ降低１４０ｋＤ、８０ｋＤ、７０ｋＤ、５６ｋＤ４种动粒蛋白的含量，ＨＴ作用２４ｈ后，几乎检测不到１４０ｋＤ和７０ｋＤ２种动粒蛋白的存在，８０ｋＤ和５６ｋＤ２种动粒蛋白的含量也急剧下降。而５２ｋＤ和１７ｋＤ２种动粒蛋白的含量则保持相对稳定。（３）ＨＴ抑制ＨＬ６０细胞中ＣＥＮＰ－Ｂ基因的表达，在对照细胞中，ＣＥＮＰ－Ｂ基因表达水平较高，ＨＴ作用６ｈ后，ＣＥＮＰ－Ｂ基因的表达就受到严重抑制，表达水平急剧下降。结果表明，ＨＴ处理能够导致某些动粒蛋白含量的降低，而且这种作用可能是通过抑制基因表达来?
Kinetochore is an important organlle in cell division. Kinetochore/Centromere is composed of protein complex. The aim of this study is to explore the relationship between the effects of anti leukemia drug harringonine(HT) on the HL60 cell and its centromere proteins by methods of indirect immunofluorecence, Western blot, Nothern blot and dot blot. We got the following results:(1) Indirect immunofluorescence shows that the brightness of kinetochore fluorescence decreases with HT treatment, which indicates the contents of centromere proteins can be reduced by HT.(2)Western blot shows that the anticentromere antibody(ACA)serum can recognize six kinds of centromere proteins with different molecular weight:140kD,80kD,70kD,56kD,52kD and 17kD. The contents of the former four kinds can be reduced by HT, while those of the latter two kinds remain relative constant. (3) Northern blot and dot blot show that the level of CENP B mRNA in HT treated cells is much less than that in control cells. These results suggest that HT can induce the decrease of the intracellular level of some centromere proteins, probably by inhibiting the gene expressions of these proteins.
出处
《解剖学报》
CAS
CSCD
北大核心
1998年第2期190-194,共5页
Acta Anatomica Sinica
基金
四川省教委重点课题资助
国家自然科学基金
