多壁碳纳米管致人肝癌细胞HepG2毒性及代谢酶表达变化

赵琢; 柳明; 杨磊; 刘伟; 王华; 刘美玲; 张秀霞; 申敬; 张园

多壁碳纳米管致人肝癌细胞HepG2毒性及代谢酶表达变化

赵琢¹,
柳明¹,
杨磊¹,
刘伟¹,
王华¹,
刘美玲²,
张秀霞²,
申敬²,
张园^1,

1.
天津海关工业产品安全技术中心, 天津 300308;
2.
天津科技大学生物工程学院, 天津 300457

基金项目: 海关总署科研计划项目（2018IK004）.

详细信息

作者简介:
赵琢,博士,研究员.E-mail:zhaozhuo@customs.gov.cn

通讯作者:
张园,博士,研究员.E-mail:zhangyuan3@customs.gov.cn

中图分类号: TQ127.1⁺1
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- 被引次数: 0
出版历程
- 收稿日期: 2019-09-29
- 录用日期: 2020-01-03
- 修回日期: 2019-12-03
- 刊出日期: 2019-12-28

The effects of multi-wall carbon nanotubes on the toxicity and the expression of metabolic enzymes (GSTP1 and CYP1A1) in human liver cancer HepG2 cells

1.
Tianjin Customs Technical Center for Safety of Industrial Products, Tianjin 300308, China;
2.
College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China

Funds: General Administration of Customs Science and Technology Project (2018IK004).

摘要

摘要: 采用实时无标记细胞分析系统研究多壁碳纳米管（MWCNTs）对人肝癌细胞HepG2毒性作用，确定MWCNTs是否会影响肝脏代谢酶谷胱甘肽S转移酶P1（GSTP1）和细胞色素P450亚酶CYP1A1的表达。利用0.12、0.25、0.5、1和2 mg/mL的MWCNTs处理HepG2细胞24、48和72h后，采用噻唑蓝比色法（MTT）测定MWCNTs对肝癌细胞HepG2增殖的抑制作用。利用实时无标记动态细胞分析（RTCA）系统动态监测不同剂量（0.25、0.5、1 mg/mL）MWCNTs对HepG2细胞生长的影响。细胞经MWCNTs处理48 h后，实时定量荧光PCR（RT-PCR）和蛋白质印迹法（Western Blot）检测肝脏代谢酶GSTP1和CYP1A1的mRNA水平和蛋白水平的变化情况。结果显示MWCNTs可以抑制HepG2细胞的增殖，且呈一定的剂量和时间依赖性。实时无标记细胞分析法能够准确监测MWCNTs对细胞增殖的过程，并且能避免多壁碳纳米管材料对数据的干扰。经MWCNTs处理后的细胞中肝脏代谢酶GSTP1mRNA水平和蛋白水平上调，而CYP1A1的mRNA水平和蛋白水平显著下降。
- 多壁碳纳米管 /
- HepG2细胞 /
- 毒性研究 /
- 实时无标记细胞分析技术 /
- 肝脏代谢酶
Abstract: To investigate the toxic effect of multiwall carbon nanotubes (MWCNTs) on the human hepatoma cell line HepG2 and to determine whether MWCNTs may affect the expression of metabolic enzymes GSTP1 and CYP1A1in HepG2, HepG2 cells were treated with 0.12, 0.25, 0.5, 1 and 2 mg/mL MWCNTs for 24, 48 and 72 h, and the effect of MWCNTs on the proliferation of HepG2 cells was then measured by thiazolyl blue colorimetry. Real-time cell analysis (RTCA), which accurately monitored the inhibitory effect on the proliferation of breast cancer MCF-7 cells with MWCNT treatment and also avoided interference of MWCNTs with the data, was used to monitor the proliferation of HepG2 cells treated with different doses of MWCNTs (0.25, 0.5, 1 mg/mL). After 48 h of treatment with MWCNTs, the mRNA and protein level of metabolic enzymes in liver were detected by a real-time polymerase chain reaction and western blotting. Results indicated that MWCNTs inhibited the proliferation of HepG2 cells in a dose and time dependent manner. The mRNA level and protein level of the metabolic enzyme GSTP1 in HepG2 increased while the mRNA level and protein level of CYP1A1 in HepG2 decreased in the cells treated by MWCNTs compared with the control group.
- Multiwalled carbon nanotube /
- HepG2 cells /
- Toxicity study /
- Real-time cell analysis /
- Liver metabolic enzyme

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