ZGDHu-1-inducing apoptosis of SHI-1 leukemia cells and its molecular mechanism.
- Author:
Yong-Lie ZHOU
1
;
Ya-Ping LÜ
;
Wei-Xiao HU
;
Lian-Nü QIU
;
Wen-Song WANG
;
Jian-Dong LIU
;
Jian-Guo WU
Author Information
1. Central Laboratory, Zhejiang Provincial People Hospital, Hangzhou 310014 China. ZYL@126.com
- Publication Type:Journal Article
- MeSH:
Animals;
Antineoplastic Agents;
pharmacology;
Apoptosis;
drug effects;
Cell Line, Tumor;
Dose-Response Relationship, Drug;
Heterocyclic Compounds, 1-Ring;
pharmacology;
Humans;
Leukemia, Monocytic, Acute;
pathology;
Mice;
Mice, Nude;
Proto-Oncogene Proteins c-bcl-2;
metabolism;
RNA, Messenger;
metabolism;
Telomerase;
metabolism;
Tumor Suppressor Protein p53;
metabolism;
bcl-2-Associated X Protein;
metabolism
- From:
Journal of Experimental Hematology
2007;15(3):483-489
- CountryChina
- Language:Chinese
-
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 concentrations 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 transmembrane potential (Delta Psi m) were measured by dihydrorhodamin 123, and expressions 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 transcriptional 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 apoptosis was confirmed by typical cell morphology, DNA fragmentation, sub-G(1) 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 Delta Psi m 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 transmembrane 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.
