[氯化镉对HepG2细胞系DNA损伤及gadd基因表达水平的影响]

[The effects of the cadmium chloride on the DNA damage and the expression level of gadd gene in HepG2 cell line].

作者信息

Zhang Rong, Niu Yu-Jie, Zhang Zhen-Jie, Chen Qing, Guo Hui-Cai, Zhao Juan, Li Yao, Fan Long-Gang

机构信息

Department of Occupational and Environmental Health, School of Public Health, Hebei Medical University, Shijiazhuang 050017, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2011 Jun;29(6):409-12.

PMID:22096849

Abstract

OBJECTIVE

To investigate the effects of the cadmium chloride on the DNA damage and the expression of the gadd153 and gadd45beta promoter and mRNA in HepG2 cells.

METHODS

DNA damage induced by cadmium chloride was detected by comet assay. The plasmids (pGADD153-Luc and pG45-Luc) containing DNA damage and repair inducible gene 153 and 45 (gadd153 and gadd45beta) promoter and luciferase and gadd45beta reporter gene were constructed. The activity of gadd153 and gadd45beta promoter were represented by the luciferase activity, the inducible luciferase activities was detected by bioluminescence. The expression of gadd153 and gadd45beta mRNA was detected by RT-PCR.

RESULTS

The results of comet assay indicated that Olive Tail Moment induced by the cadmium chloride increased significantly at the dose of 100, 300 micromol/L, compared with the control (P < 0.05). The luciferase activity analysis showed that the expression levels of gadd153 promoter increased significantly in 1, 5, 10 micromol/L treatment group, compared with the control (P < 0.05). The expression levels of gadd45beta promoter in 5, 10 micromol/L treatment group were significantly higher than that in control group (P < 0.05). The expression levels of gadd153 mRNA induced by cadmium chloride at the doses of 1, 5, 10 micromol/L and the expression levels of gadd45beta mRNA induced at the doses of 5, 10 micromol/L were significantly higher than thoae in control group (P < 0.05).

CONCLUSION

The cadmium chloride can induce the DNA damage and increase the expression levels of the gadd153 and gadd45beta promoters in HepG2 cells.

摘要

目的

研究氯化镉对HepG2细胞DNA损伤以及生长停滞和DNA损伤诱导基因153（gadd153）和生长停滞和DNA损伤诱导基因45β（gadd45β）启动子及信使核糖核酸（mRNA）表达的影响。

方法

采用彗星试验检测氯化镉诱导的DNA损伤。构建含有DNA损伤与修复诱导基因153和45（gadd153和gadd45β）启动子、荧光素酶及gadd45β报告基因的质粒（pGADD153-Luc和pG45-Luc）。以荧光素酶活性代表gadd153和gadd45β启动子活性，通过生物发光检测诱导的荧光素酶活性。采用逆转录-聚合酶链反应（RT-PCR）检测gadd153和gadd45β mRNA的表达。

结果

彗星试验结果表明，与对照组相比，100、300μmol/L剂量的氯化镉诱导的橄榄尾矩显著增加（P<0.05）。荧光素酶活性分析显示，与对照组相比，1、5、10μmol/L处理组中gadd153启动子的表达水平显著增加（P<0.05）。5、10μmol/L处理组中gadd45β启动子的表达水平显著高于对照组（P<0.05）。1、5、10μmol/L剂量的氯化镉诱导的gadd153 mRNA表达水平以及5、10μmol/L剂量诱导的gadd45β mRNA表达水平均显著高于对照组（P<0.05）。