铅和镉对人胎盘绒毛癌细胞中S100钙结合蛋白成员mRNA表达的影响

doi:10.3969/j.issn.1004-616x.2012.03.002

›› 2012, Vol. 24 ›› Issue (3): 166-171.doi: 10.3969/j.issn.1004-616x.2012.03.002

铅和镉对人胎盘绒毛癌细胞中S100钙结合蛋白成员mRNA表达的影响

朱敏,张庆英,罗家逸,张东红,赵治国,易德青

1. 汕头大学医学院预防医学教研室，广东汕头 515041；2. 汕头大学医学院心血管研究中心，广东汕头 515041

收稿日期:2011-12-20 修回日期:2012-02-29 出版日期:2012-05-30 发布日期:2012-05-30
通讯作者: 张庆英

The effect of lead and cadmium on mRNA of S100 calcium-binding protein family members in JAR cells

ZHU Min，ZHANG Qing-ying，LUO Jia-yi，ZHANG Dong-hong，ZHAO Zhi-guo，YI De-qing

1. Department of Preventive Medicine, Shantou University Medical College, Shantou 515041; 2. Cardiovascular Research Center, Shantou University Medical College, Shantou 515041, Guangdong, China

Received:2011-12-20 Revised:2012-02-29 Online:2012-05-30 Published:2012-05-30
Contact: ZHANG Qing-ying

摘要/Abstract

摘要： 目的：探讨重金属铅 (Pb)和镉 (Cd)对人胎盘绒毛癌细胞 (JAR)中一组S100钙结合蛋白家族成员mRNA表达的影响。方法：采用四甲基噻唑蓝 (MTT)实验分别检测乙酸铅 (PbAc)和乙酸镉 (CdAc)对JAR增殖活力的影响。再进一步采用小于半数致死量的不同浓度的PbAc和CdAc分别染毒JAR细胞24 h和48 h，运用半定量RT-PCR检测各染毒组与对照组[金属硫蛋白 (MT)为参照，β-actin为内参]S100钙结合蛋白家族成员mRNA的表达水平。结果：PbAc (2.5～20 μmol/L)和CdAc (2.5～10 μmol/L)染毒24 h后对JAR细胞的相对存活率与染毒浓度呈剂量效应关系，且呈线性负相关 (r 依次为-0.992 4和-0.995 5，P均 < 0.05)，半数致死浓度PbAc为23.15 μmol/L， CdAc 为8.30 μmol/L。半定量RT-PCR检测9个钙结合蛋白S100家族成员，仅发现S100A10、S100A11、S100A14和S100P有明显表达，其余的S100成员表达较弱或未能检出。染毒24 h和48 h后，与对照组比较，S100A10和S100A11 mRNA的表达未发生明显变化，但S100A14 mRNA的表达 (除10 μmol/L PbAc染毒24 h外)明显上调 (P <0.05或P <0.01)。S100P的表达经2.5和5 μmol/L PbAc染毒24 h后明显上调 (P <0.05或P <0.01)，但随PbAc浓度增高表达有降低的趋势。而经CdAc染毒24 h后各实验组S100P的表达均明显上调 (P <0.05 或P <0.01)；经染毒48 h后的表达进一步增加，但与对照组相比差异无统计学意义 (P >0.05)。S100P的表达与MT对照的结果相类似。结论：PbAc或CdAc染毒JAR细胞可引起S100A14和S100P mRNA的表达发生明显的变化，其中S100P的变化规律与MT的表现类似，推测这两种钙结合蛋白在重金属毒作用过程中有参与保护细胞的重要作用，可作为细胞应激的分子生物指标。

关键词: 乙酸铅, 乙酸镉, 人胎盘绒毛癌细胞, S100钙结合蛋白

Abstract: OBJECTIVE: To explore the effects of lead acetate (Pb) and cadmium acetate (Cd) on the mRNA expression of S100 calcium-binding protein family members in JAR cells. METHODS：MTT Viability Test was performed to determine the median lethal dose (LC50) in JAR cells. JAR cells were treated by different concentrations of PbAc and CdAc for 24 h and 48 h. Expressions of S100 proteins mRNA by RT-PCR between different experimental groups and control group was compared. RESULTS：After PbAc (2.5-20 μmol/L) and CdAc (2.5-10 μmol/L) treatmeat of JAR cells，the relative cell viability showed a negative correlation with concentration (r = -0.992 4 or -0.995 5，P < 0.05 for both). The median lethal concentration (LC50) of PbAc was 23.15 μmol/L，and that of CdAc was 8.30 μmol/L. Among the nine S100 calcium-binding protein family members，only S100A10， S100A11， S100A14 and S100P were found to be significantly expressed，while others showed lower expression or no expression by RT-PCR. The mRNA expression of S100A14 in JAR cells treated with PbAc and CdAc after 24 h and 48 h were significantly different from the negative control (P <0.05)，while no significant difference was found for the expression of S100A1 and S100A11. When JAR cells were treated with 2.5 μmol/L PbAc for 24 h，the expression of S100P significantly increased (P <0.05)，but expression declined as the dose of PbAc increased. When JAR cells were treated with CdAc for 24 h，the expression of S100P significantly increased (P <0.05). However after the exposure of PbAc and CdAc for 48 h，S100P expression of all experimental groups were increased，with no significant difference compared with the control group. CONCLUSION：PbAc and CdAc could increase the mRNA expressions of S100A14 and S100P in JAR cells，and the variation of S100P expression was similar to metallothionein (MT). The enhanced mRNA expressions of S100A14 and S100P may be involved in heavy metal toxic stimulation stress response. We postulate that they may protect cells from the toxic effects of heavy metals，and they might be molecular biomarkers for stress response.

Key words: lead acetate, cadmium acetate, JAR cells, S100 calcium-binding protein

中图分类号:

R734.2

朱敏,张庆英,罗家逸,张东红,赵治国,易德青. 铅和镉对人胎盘绒毛癌细胞中S100钙结合蛋白成员mRNA表达的影响[J]. , 2012, 24(3): 166-171.

ZHU Min，ZHANG Qing-ying，LUO Jia-yi，ZHANG Dong-hong，ZHAO Zhi-guo，YI De-qing. The effect of lead and cadmium on mRNA of S100 calcium-binding protein family members in JAR cells[J]. , 2012, 24(3): 166-171.

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铅和镉对人胎盘绒毛癌细胞中S100钙结合蛋白成员mRNA表达的影响

The effect of lead and cadmium on mRNA of S100 calcium-binding protein family members in JAR cells

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