Publication Date:
2016-12-02
Description:
Objective: To observe the mechanisms of realgar intervening telomere protein POT1, TRF1, TRF2 expression to regulate telomere dynamics in THP-1 cells, and elucidate the experimental and theoretical basis for realgar targeted threapy of AML. Methods: 1.In vitro: (1)Cultured human acute myeloid leukemia THP-1 cells. (2), The cells were incubated in absence or presence of increasing concentration of realgar for 24/48 h, from which determined the IC50. Cell viability was tested by CCK-8 assay. (3)Apoptotic rate and cell cycle distribution were tested by FCM. (4)Each group of POT1, TRF1, TRF2 protein levels were tested by western-blot analysis. (5)Each group of POT1,TRF1,TRF2 mRNA levels were tested by RT-qPCR analysis. 2.In vivo: (1)Established THP-1 model in NOD/SCID mice, and the mouse were treated by realgar. (2)All the mouse were killed by institutionally approved method after 3 weeks. The apoptotic rate and cell cycle distribution of mice spleen cells were examined by FCM. (3)The changes of POT1, TRF1, TRF2 protein levels in organic tissue were detected by IHC. Results: 1.In vitro: (1)CCK-8 assay showed that after treated by realgar, THP-1 cells growth were inhibited and the cell viability decreased in a dose- and time- dependent manner, IC50 was 0.023μg/mL. Therefore, we choose a medium concentration was 0.015μg/mL (lower than IC50) at 48h. (2)Our study demonstrated that after treatment for 48h, the apoptotic rate and the G1 arrest of these THP-1 cell increased, compared with control group. (3)As shown by western blot analysis, compared with controls, realgar group POT1, TRF1 protein levels significantly increaesed and TRF2 decreased(P
Print ISSN:
0006-4971
Electronic ISSN:
1528-0020
Topics:
Biology
,
Medicine
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