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  

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

作　　者：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] 

机构地区：[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

出　　处：《吉林大学学报（医学版）》2010年第5期847-850,共4页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金青年基金资助课题(30700672)

摘　　要：目的:观察氯化镉(CdCl2)处理人肝癌SMMC-7721细胞系中CHOP和GRP78mRNA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40μmol·L-1的培养液中24h,0μmol·L-1组为对照组。采用SYBR荧光实时定量PCR法检测CHOP和GRP78mRNA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,CHOPmRNA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40μmol·L-1组与对照组比较差异有显著性(P<0.05);GRP78mRNA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40μmol·L-1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GRP78mRNA表达增多。Objective To study the effects of cadmium chloride （CdCl2）on endoplasmic reticulum stress by observing the expressions of CHOP and GRP78mRNA in SMMC-7721cells.Methods SMMC-7721cell cultures were exposed to CdCl2at concentrations of 0,5,10,20,30and 40μmol·L-1 for 24h.Power SYBR green PCR Master Mix and Mx3000PQPCR 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 CdCl2dose.The expressions of CHOP mRNA in all experimental groups were higher than that in control group,and the expressions of CHOP mRNA in 10,20and 40μmol·L-1 groups were increased significantly（P〈0.05）.The GRP78mRNA level was increased along with the elevating of CdCl2dose.The expressions of GRP78mRNA in 5,20and 40μmol·L-1 groups were significantly higher than that in control group（P〈0.05or P〈0.01）.Conclusion CdCl2can induce the expressions of CHOP and GRP78 mRNA in SMMC-7721cells.

关 键 词：氯化镉 SMMC 7721 CHOP 葡萄糖调节蛋白78 

分 类 号：R994.6[医药卫生—毒理学]