实时定量PCR检测氯化镉对人肝癌细胞SMMC-7721中CHOP和GRP78 mRNA表达的影响

J4 ›› 2010, Vol. 36 ›› Issue (5): 847-850.

实时定量PCR检测氯化镉对人肝癌细胞SMMC-7721中CHOP和GRP78 mRNA表达的影响

刘颖¹, 孙世龙², 刘晓梅¹, 杜海英¹, 李鹏¹, 杜忠君¹, 孙磊¹, 李阳¹, 徐艳玲¹, 孙志伟^1,3

1.吉林大学公共卫生学院卫生毒理学教研室,吉林长春 130021；2.吉林大学公共卫生学院卫生部放射生物学重点实验室|吉林长春 130021；3.首都医科大学公共卫生学院卫生毒理学教研室|北京 100054

收稿日期:2010-06-17 出版日期:2010-09-28 发布日期:2010-09-28
通讯作者: 孙志伟(Tel：010-83911507，E-mail：zwsun@hotmail.com) E-mail:zwsun@hotmail.com
作者简介:刘颖(1978-),女,黑龙江省兰西县人|讲师|医学博士,主要从事环境毒理学研究。
基金资助:
国家自然科学基金青年基金资助课题(30700672) 

Effects of cadmium chloride on expressions of CHOP and GRP78 　mRNA in |SMMC-7721 cells detected with real time quantitative PCR

LIU Ying¹, SUN Shi-Long², LIU Xiao-Mei¹, DU Hai-Ying¹, LI Peng,¹ DU Zhong-Jun¹,SUN Lei¹, LI Yang¹, XU Yan-Ling¹, SUN Zhi-Wei^1,3

1. Department of Toxicology,School of Public Health,Jilin University,Changchun 130021,China;2. Key Laboratory of Radiobiology,Ministry of Health, School of Public Health,Jilin University,Changchun 130021,China;3. Department of Toxicology,School of Public Health,Capital Medical University,Beijing 100054,China

Received:2010-06-17 Online:2010-09-28 Published:2010-09-28

摘要/Abstract

摘要：

目的：观察氯化镉（CdCl2）处理人肝癌SMMC-7721细胞系中CHOP和GRP78 mRNA表达的改变，在mRNA水平探讨氯化镉对肝癌细胞内质网应激的影响。方法：以人肝癌细胞株SMMC-7721细胞为研究模型，细胞暴露于氯化镉浓度为0、5、10、20、30及40 μmol?L-1的培养液中24 h，0 μmol/L组为对照组。采用SYBR荧光实时定量PCR法检测CHOP和GRP78 mRNA表达，相对定量采用比较CT值法。结果：随着氯化镉剂量的增加，CHOP mRNA表达水平有逐渐增多的趋势，各组mRNA表达均高于对照组，其中10、20和40 μmol/L组与对照组比较差异有显著性（P<0.05）；GRP78 mRNA表达有逐渐增多的趋势，各组mRNA表达均高于对照组，其中5、20和40 μmol/L组与对照组比较差异有显著性（P<0.05或P<0.01）。结论：氯化镉可诱导人肝癌SMMC-7721细胞中CHOP和GRP78 mRNA表达增多。

关键词: 氯化镉；SMMC-7721；CHOP；葡萄糖调节蛋白78

Abstract:

Abstract:Objective To study the effects of cadmium chloride (CdCl2) on endoplasmic reticulum stress by observing the expressions of CHOP and GRP78 mRNA in SMMC-7721 cells. Methods SMMC-7721 cell cultures were exposed to CdCl2 at concentraions of 0,5,10,20,30 and 40 μmol /L for 24 h. Power SYBR green PCR Master Mix and Mx3000P QPCR System were used to measure a threshold cycle (CT) value for each sample. Relative quantitation of gene expression was based on the comparative CT method. Results The CHOP mRNA level tended to increase along with the elevating of CdCl2 dose. The expressions of CHOP mRNA in all experimental groups were higher than that in control group,and the expressions of CHOP mRNA in 10,20 and 40 μmol /L groups were increased significantly (P<0.05). The GRP78 mRNA level was increased along with the elevating of CdCl2 dose. The expressions of GRP78 mRNA in 5,20 and 40 μmol /L groups were significantly higher than that in control group (P<0.05 or P<0.01). Conclusion CdCl2 can induce the expressions of CHOP and GRP78 mRNA in SMMC-7721 cells.

Key words: cadmium chloride;SMMC-7721;CHOP;glucose-regulated protein of 78 kDa

中图分类号:

R994.6

刘颖, 孙世龙, 刘晓梅, 杜海英, 李鹏, 杜忠君, 孙磊, 李阳, 徐艳玲, 孙志伟. 实时定量PCR检测氯化镉对人肝癌细胞SMMC-7721中CHOP和GRP78 mRNA表达的影响[J]. J4, 2010, 36(5): 847-850.

LIU Ying, SUN Shi-Long, LIU Xiao-Mei, DU Hai-Ying, LI Peng, DU Zhong-Jun, SUN Lei, LI Yang, XU Yan-Ling, SUN Zhi-Wei. Effects of cadmium chloride on expressions of CHOP and GRP78 　mRNA in |SMMC-7721 cells detected with real time quantitative PCR[J]. J4, 2010, 36(5): 847-850.