5-aza-2′-deoxycytidine suppresses the growth of human lung adenocarcinoma cells in nude mouse xenograft models and its effect on methylation status and expression of TFPI-2 gene
10.3760/cma.j.issn.0253-3766.2015.07.002
- VernacularTitle:5-氮杂-2′-脱氧胞苷对人肺腺癌裸鼠种植瘤的抑制作用及组织因子途径抑制物2基因甲基化状态和表达的影响
- Author:
Jiangshui LIANG
1
;
Guilin YIN
;
Yongqiang DONG
;
Zhongsha MA
;
Yuehua XIAO
;
Tao JI
Author Information
1. 423000,郴州市第一人民医院胸部肿瘤肺癌诊疗中心
- Keywords:
Lung neoplasms;
Adenocarcinoma;
Mice;
DNA methylation;
5-aza-2′-deoxycytidine;
Tissue factor pathway inhibitor 2
- From:
Chinese Journal of Oncology
2015;(7):485-489
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effect of classic demethylating drug 5?aza?2′?deoxycytidine( 5?Aza?CdR ) on the growth of human lung adenocarcinoma cells in nude mouse xenograft models, and to observe its effect on methylation status and expression of TFPI?2 gene in the nude mouse xenograft tissues. Methods The nude mouse xenograft model was established by subcutaneous inoculation of human lung adenocarcinoma A549 cells.According to different doses of 5?Aza?CdR, the tumor?bearing nude mice were randomly divided into experimental groups (0.5 mg/kg group, 1 mg/kg group, 2 mg/kg group) and control group (0 mg/kg group). The tumor growth in the nude mice was observed. The methylation status and the expression of TFPI?2 gene mRNA and protein were detected by methylation specific polymerase chain reaction, real?time fluorescent quantitative polymerase chain reaction and Western blot assay. Results The nude mice were euthanized at 28 days after intraperitoneal injection of 5?Aza?CdR. The body weight of tumor?bearing nude mice was (27.12±0.38)g in the 0 mg/kg group, (26.80±0.18)g in the 0.5 mg/kg group, (26.67±0.28)g in the 1 mg/kg group, and (26.50±0.26)g in the 2 mg/kg group, showing no significant difference among them (P>0.05). The volume of xenograft tumors in the 0 mg/kg group was (709.22±2.87)mm3, (400.67±2.68)mm3 in the 0.5 mg/kg group, (285.71±2.91)mm3 in the 1 mg/kg group, and (230.44 ±3.15)mm3 in the 2 mg/kg group, showing a significant difference (P<0.05). There were complete methylation of TFPI?2 gene in the 0 mg/kg group, incomplete methylation in the 0.5 and 1 mg/kg groups, and unmethylation in the 2 mg/kg group. The relative mRNA level in the 0, 0.5, 1, 2 mg/kg groups were 1.00±0.00, 1.67±0.07, 3.40±0.24, and 5.55±0.61, respectively (P<0.05). The relative expression level of TFPI?2 protein in the 0, 0.5, 1, 2 mg/kg groups was 0.18±0.02, 0.36±0.01, 0.64±0.02, and 0.81±0.20, respectively (P<0.05). Conclusions 5?Aza?CdR suppresses the tumor growth of human lung adenocarcinoma cells in nude mouse xenograft models, and induces expression of TFPI?2 gene in the xenograft tumor cells. The mechanism might be that 5?Aza?CdR induces re?expression of demethylated TFPI?2 gene by demethylation, and thus inhibits the growth and proliferation of human lung adenocarcinoma cells.
