| External factors: | Triptonide |
| Aging type: | Accelerate |
| Aging characteristic: |
| Category: | Chemical compounds |
| Phenotype: | Acute leukemia |
| Experiment: | Flow cytometry//Cell morphological analysis//SA-β-gal activity assay |
| Description: | Cell cycle assay showed that the cell populations at G0/G1 phases were significantly increased for 40.6% at the dose of 10 nM, but those at G2/M phases were significantly decreased for 38% after 3 days of triptonide treatment, suggesting that triptonide increases leukemia cell arrest at G0/G1.In particular, cell size was enlarged, and cells profoundly changed shape from rounded to an irregular form, with budding evident on the plasma membrane surface, and an enlarged nucleus containing big vacuoles.The results showed that β-Gal-positive HL60 and U937 cells were significantly increased after 6 days of triptonide treatment by 21.3% and 25%, respectively , compared to that in the control cells. |
| Target gene: | TERT//C-MYC//P16//P21//DDIT3 |
| R-EF-Target gene: | Downregulation//Downregulation//Upregulation//Upregulation//Upregulation |
| Official symbol(s): | TERT//C-MYC//P16//P21//DDIT3 |
| Target gene experiment: | RT-PCR//Western blot |
| Target gene description: | QT -PCR analysis indicated that TERT expression in HL60 cells was reduced for 6.7 and 11.4 times at triptonide doses of 10 and 15 nM, respectively. In addition, western blotting analysis showed that TERT protein levels were also significantly diminished .RT -PCR analysis showed that the mRNA levels of p16 were significantly increased, while p21 in the cells were notably elevated, and QT-PCR analysis indicated that the expression levels of p16 and p21 were increased by 9- and 24.7-fold, respectively , after 3 days of treatment with 15 nM triptonide, respectively.The results showed that c-Myc gene expression was signif icantly reduced in the presence of 15 nM triptonide, whereas, triptonide at doses of 10 nM and less did not significantly promote c-Myc expression, RT -PCR analysis showed that triptonide treatment markedly elevated DDIT3 mRNA levels.Western blotting indicated that DDIT3 protein levels were significantly elevated in HL-60 cells after triptonide treatment. |
| Regulatory pathway: | MKK3-p38//ERK |
| R-EF-Pathway: | Upregulation//Downregulation |
| Official symbol(s): | MAP2K3-MAPK14//ERK |
| Pathway experiment: | Western blot |
| Pathway description: | Western blotting indicated that the level of phsphorylated p38 was significantly increased, whereas the level of p-ERK was obviously diminished . |
Annotation: