Wu Dong, Zhang Yao, Zhao You-Shan, Guo Juan, Chang Chun-Kang
Department of Hematology, The Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 200233, China.
Department of Hematology, The Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 200233, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Oct;25(5):1471-1476. doi: 10.7534/j.issn.1009-2137.2017.05.033.
To investigate the inhibitory effect of decitabine (DAC) in various dosages on the proliferention of MDS-RAEB cell line MDS-L and its mechanism.
LC-MS/MS method was used to test the blood DAC concentration of 2 groups of MDS patients being treated with DAC 20 and 15 mg/m×5 d. In according to the various blood DAC concentration levels, the MDS-L cells were treated with different DAC dosages for 24, 48, 72 and 96 h, respectively. The CCK-8 method was applied to determine the cell proliferation, the flow cytometry was used to analyze the cell cycle and cell apoptosis changes, the P15 DNA methylation status was measured by methylation specific PCR using EZ DNA Methylation-Gold Kit.
The blood DAC concentration of MDS patients treated with DAC 20 mg/m×5 d was 174.08±80.15(84.7-311) ng/ml, which was significantly higher than 89.87±32.94(43.2-165)ng/ml for the group treated with 15 mg/m×5 d (P=0.014). DAC could notably inhibit the proliferation of MDS-L cells, and the effect was in dose- and- time-dependent manner(r=0.786). However, when DAC concentration was ≥0.1 µg/ml, the proliferation inhibition rates were not significantly different between various dosages. After DAC treatment, MDS-L cells in G phase increased notably, while cells in S phase decreased significantly. Also, the P15 DNA methylation status of MDS-L cells decreased after being treated with DAC for 96 h, but the difference was not significant between various dosages.
DAC can significantly suppress MDS-L cell proliferation, block MDS-L cells in G phase and induce the apoptosis at low concentration (0.1-0.2 µg/ml).
