Tetrazine dimethyl amide-induced leukemia cell line SHI 1 and its molecular mechanism of apoptosis.docVIP

 PAGE \* MERGEFORMAT 32 Tetrazine dimethyl amide-induced leukemia cell line SHI 1 and its molecular mechanism of apoptosis Author: Zhou columns, Ya-Ping Lü, Wei-Xiao Hu, Lian-Nü Qiu, Wen-Song Wang, Jian-Dong Liu, Wu Jianguo Abstract This study investigated the tetrazine-dimethyl amide (ZGDHu  1)-induced leukemia cell line SHI  1 apoptosis mechanism. With different concentrations of ZGDHu  1 in vitro SHI  1 cells, using cell morphology, DNA gel electrophoresis, DNA content and cell cycle analysis, Annexin  V / PI double-labeled and fluorescent staining and other analysis Hoechst33258 apoptosis. Rh123/PI measured by mitochondrial transmembrane potential (ΔΨm), using flow cytometry ZGDHu  1 induced by SHI  1 cell apoptosis bcl  2, bax, p53, Fas protein and mitochondrial membrane protein expression. By RT  PCR observe the role of ZGDHu  1, after SHI  1 cells bcl  2, bax, p53 gene expression changes; while taking advantage of real-time fluorescence quantitative PCR detection of telomerase hTERT  mRNA expression changes. The results showed that: ZGDHu  1 can induce apoptosis of SHI  1, showing the amount of the role of time-and dose-effect relationship, the typical morphological changes, DNA fragmentation, sub-G1 peak detected and significantly increased; Annexin  V / PI and Hoechst33258 fluorescence staining cells characteristic changes of apoptosis. SHI  1 cells were treated with different concentrations of ZGDHu  1 in vitro cultured, bcl  2 gene and protein has been reduced, but mainly bax gene and protein increases, leading to bax / bcl  2 were significantly increased, p53 gene and protein and Fas protein expression also increases, Jun Cheng dose-dependent. With ZGDHu  1 role in the increase in the concentration, ΔΨm decreased, while the mitochondrial membrane protein expression significantly increased the expression of telomerase hTERT  mRNA significantly reduced. Conclusion: ZGDHu  1 by upregulating the p53 gene, bax gene and