亚砷酸钠对人皮肤永生化角质形成细胞角化相关基因和核转录因子红系相关因子2mRNA表达的影响  

Effects of sodium arsenite on mRNA transcription of keratinizing related and nuclear factor E2-related factor 2 genes in HaCaT cells

作　　者：胡新欣[1] 高彦辉[1] 张微[1] 孙惠昕[1] 孙殿军[1] 

机构地区：[1]哈尔滨医科大学中国疾病预防控制中心地方病控制中心地氟病防治研究所卫生部病因重点实验室、黑龙江省教育厅病因流行病学重点实验室,150081

出　　处：《中国地方病学杂志》2012年第4期365-368,共4页Chinese Jouranl of Endemiology

基　　金：“十一五”国家科技支撑项目（2006BA106804）

摘　　要：目的观察不同浓度亚砷酸钠（NaAsO2）对人皮肤永生化角质形成细胞（HaCaT）角化相关基因和核转录因子红系相关因子2（nuclear factor erythroid 2-related factor2，Nrf2）mRNA表达的影响。方法用0．00（对照）、3．13、6．25、12．50、25．00、50．00、75．00、100．00p．mol／LNaAs02处理HaCaT细胞48h，采用2-（2-甲氧基-4-硝基苯基）-3-（4-硝基苯基）-5-（2，4-二磺酸苯）-2H-四唑单钠盐（CCK-8）法检测细胞增殖率。根据细胞增殖率检测结果，选择0．00（对照）、6．25、12．50、25．00μmol／LNaAs02处理HaCaT细胞48h，采用实时荧光定量PCR法检测HaCaT细胞角蛋白1（Keratinl，K-1）、角蛋白10（Keratin 10，K-10）、总苞蛋白（Involurin，Inv）、兜甲蛋白（Loricfin，Lor）和N以的mRNA表达。结果与对照组（100．05％）比较。6．25、12．50、25．00、50．00、75．00、100．00μmol／LNaAsO2组HaCaT细胞增殖率（83．06％、51．04％、39．52％、24．51％、16．99％、9．04％）明显降低，半数致死量为12．38μmol／L。与对照组（1．06±0．28、1．00±0．12、1．00±0．08）比较，6．25、12．50、25．00μmol／LNaAsO2组HaCaT细胞K—1、Inv、LormRNA表达（0．08±0．04、0．13±0．12、0．05±0．03，0．47±0．11、0．21±0．09、0．10±0．15，0．50±0．27、0．31±0．10、0．57±0．23）明显降低（P均〈0．05），但K-10mRNA表达呈现升高趋势，其中6．25μmol／LNaAs02组（1．83±0．45）明显高于对照组（1．07±0．14，P〈0．05）；而12．50、25．00μmol／LNaAsO2组Nrf2mRNA表达（0．13±0．07、0．69±0．33）明显低于对照组（1．00±0．09，P均〈0．05）。结论NaAsO2可通过影响细胞角化相关基因和Nrf-2mRNA表达，抑制HaCaT细胞增殖与角化。Objective To observe the influences of different doses of sodium arsenite on mRNA transcription of keratinizing related and nuclear factor E2-related factor 2（Nrf2） genes in HaCaT cells. Methods Cell proliferation was evaluated by Cell Counting Kit-8（CCK-8） assay after the HaCaT cells were exposed to 0.00, 3.13, 6.25, 12.50, 25.00, 50.00, 75.00, 100.00 μmoL/L sodium arsenite for 48 h, respectively. Based on the previous results of cell proliferation, 0.00（control）, 6.25, 12.50, and 25.00 μmol/L of sodium arsenite were selected to treat HaCaT cells for 48 h, respectively. The mRNA expression of keratin 1, keratin 10, involucrin, loricrin and Nrf2 were detected by real-time fluorescent quantitative PCR. Results Compared with the control group （100.05%）, HaCaT cell proliferation rates（83.06%, 51.04%, 39.52%, 24.51%, 16.99% and 9.04%） were significantly lower in 6.25, 12.50, 25.00, 50.00, 75.00 and 100.00 μmol/L of sodium arsenite groups and the 50% inhibiting concentration was 12.38 μmol/L. Compared with the control group（1.06 ± 0.28, 1.00 ± 0.12, 1.00 ± 0.08）, the mRNA expression of keratin 1, involucrin and loricrin （0.08 ± 0.04, 0.13 ± 0.12, 0.05 ± 0.03; 0.47 ± 0.11, 0.21 ± 0.09, 0.10 ± 0.15; 0.50 ± 0.27, 0.31 ± 0.10, 0.57 ± 0.23） were significantly decreased（all P 〈 0.05） in HaCaT cells treated with 6.25, 12.50, 25.00 μmol/L sodium arsenite, respectively. But keratin I0 mRNA expression showed a rise trend and the 6.25 μmol/L sodium arsenite group （1.83 ± 0.45） was significantly higher than that of the control（1.07 ± 0.14, P 〈 0.05）. The Nrf2 mRNA expressions of HaCaT cells in 12.50, 25.00 μmol/L sodium arsenite groups （0.13 ± 0.07, 0.69 ± 0..33 ） were significantly lower than that of＂ the control （ 1.00 ± 0.09, all P 〈 0.05 ）. Coneluslons The cellular proliferation and keratinization are decreased when HaCaT cells are exposed to sodium arsenite, which may be regulated by lowering Nrf2 mRNA transcription.

关 键 词：角蛋白1 角蛋白10 总苞蛋白 兜甲蛋白 亚砷酸盐类 NF-E2-相关因子2 

分 类 号：R599[医药卫生—内科学]