四嗪二甲酰胺诱导白血病细胞株SHI1凋亡及其分子机制研究.docVIP

四嗪二甲酰胺诱导白血病细胞株SHI1凋亡及其分子机制研究 正文 四嗪二甲酰胺诱导白血病细胞株SHI1凋亡及其分子机制研究 文1：四嗪二甲酰胺诱导白血病细胞株SHI1凋亡及其分子机制研究 Abstract The aim of study was to investigate the mechanism of N,N′di(mmethylphenyl)3,6dimethyl 1,4dihydro1,2,4,5tetrazine1,4dicarboamide(ZGDHu1) inducing apoptosis in SHI1 human leukemia cell line. Different concentratio of ZGDHu1 and different times of culture were used to treat SHI1 cells; the apoptosis of SHI1 cells was analyzed by morphology, DNA agarose gel electrophoresis, DNA content detection, AnnexinV/PI and Hoechst33258 labeling method, the mitochondrial tramembrance potential（ΔΨm） were measured by dihydrorhodamin 123, and expressio of bcl2, bax, Fas, p53 and mitochondrial membrane protein were analyzed by flow cytometry,while the bcl2,bax and p53 gene were analyzed by RTPCR. The tracriptional level of hTERTmRNA was measured by realtime fluorescence quantitative RTPCR. The results showed that after exposure to ZGDHu1, SHI1 cells were induced to apoptosis in a timeand doesdependent manner. SHI1 cell apoposis was comfirmed by typical cell morphology, DNA fragmentation, subG1 phase, Hoechst33258 and AnnexinⅤ/PI labeling etc. The expression of bax, bax/bcl2, p53 and Fas gene significantly increased and bcl2 slightly decreased. ZGDHu1 could increased the expression of mitochondrial membrane protein in a dosedependent manner while ΔΨm reduced. The expression of hTERTmRNA significantly decreased. It is concluded that ZGDHu1 can upregulate the expression of p53, bax and bax/bcl2. The mitochondrial pathway mediated by descent of mitochondrial tramembrance potential may be one of the mechanisms inducing apoptosis by ZGDHu1, in which Fas gene also participates. Telomerase may be an effective gene target for antitumour effect of ZGDHu1. Key words N,N＇di(mmethylphenyi)3,6dimethyl1,4dihydro1,2,4,5tetrazine1,4dicarboamide; cell line,SHI1; apoptosis; mitochondria;telomerase 中国 实验血液学杂志 J Exp Hematol 2007; 15(3)四嗪二甲酰胺诱导白血病细胞株SH