Effects of DNA methylation inhibitor 5-Aza-2'-deoxycytidine on biological behavior of esophageal squamous carcinoma cells
10.3760/cma.j.issn.1006-9801.2018.02.001
- VernacularTitle:DNA甲基化抑制剂5-氮杂-2'-脱氧胞苷对食管鳞状细胞癌细胞生物学行为的影响
- Author:
Lu CHANG
1
;
Yongming XU
;
Yanghui BI
;
Fang WANG
;
Hongyi LI
;
Pengzhou KONG
Author Information
1. 山西医科大学转化医学研究中心 食管癌发病机理及转化研究山西省重点实验室 细胞生理学省部共建教育部重点实验室
- Keywords:
Esophageal squamous cell carcinoma;
DNA Methylation;
5-Aza-2'-deoxycytidine;
Apoptosis;
Cell cycle
- From:
Cancer Research and Clinic
2018;30(2):73-78
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of methylation inhibitor 5-Aza-2'-deoxycytidine (5-Aza-dC) on biological behavior of esophageal squamous carcinoma cell (ESCC) lines KYSE140 and KYSE150. Methods KYSE140 and KYSE150 cell lines were divided into the blank group, the control group and the experimental group. The cells in the blank group didn't do the treatment, and the cells in the control group were added to DMSO 2 μmol/L, while in the experimental group, cells were treated with different concentration (1, 2, 3 and 4 μmol/L) of 5-Aza-dC which affected respectively at different time (24, 48, 72 and 96 h). Cell proliferation was detected by using methyl thiazolyl tetrazolium (MTT) assay and the optimal drug concentration and time point were selected. Transwell assay was performed to detect the change of cell migration and invasion. Flow cytometry was used to observe the effects of drugs on cell apoptosis and cell cycle.The expression of PARP,Caspase-3,CCNB-1,and CCNE-1 were detected by Western blot. Results MTT result showed that the effective function time of 5-Aza-dC on KYSE140 and KYSE150 was 96 h at the concentration of 4 μmol/L. Under this condition, the cell ability of migration and invasion was decreased significantly. The migrated cell number of KYSE140 and KYSE150 respectively in the blank group, the control group and the experimental group was (193.3±8.6), (184.0±10.4), (61.7±7.1) and (112.0±6.4), (101.3± 7.9), (26.3±5.7). The invasive cell number was (47.3±7.3), (38.7±5.1), (8.0±3.9) and (83.3±6.8), (74.7±5.7), (21.0±2.7), respectively. The difference was statistically significant (P <0.05). Flow cytometry revealed that 5-Aza-dC increased the apoptosis of KYSE140 and KYSE150. The apoptosis rate of the blank group, the control group and the experimental group was (2.8±0.3) %, (11.2±0.7) %, (18.6±0.6) % for KYSE140 and (2.7±0.4)%,(9.8±0.4)%,(17.7±0.5)% for KYSE150.Compared with the other two groups,the cell number of G2/M phase in the experimental group was increased remarkably (P < 0.05). PARP and Caspase-3 were sheared evidently and the protein expression of CCNB-1 was up-regulated while the expression of CCNE-1 was down-regulated in the experimental group. Conclusion 5-Aza-dC can inhibit the proliferation and promote apoptosis of ESCC cells.
