[1]贾义,钟乃凤,周静.MsrB1对ONOO－诱导的人晶状体上皮细胞周期的影响[J].眼科新进展,2015,35(11):1021-1024.[doi:10.13389/j.cnki.rao.2015.0279]
　JIA Yi,ZHONG Nai-Feng,ZHOU Jing.Effects of MsrBl on peroxynitrite-induced cell cycle arrest in human lens epithelial cells[J].Recent Advances in Ophthalmology,2015,35(11):1021-1024.[doi:10.13389/j.cnki.rao.2015.0279]
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MsrB1对ONOO－诱导的人晶状体上皮细胞周期的影响
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年11期
页码:
1021-1024
栏目:
实验研究
出版日期:
2015-11-05

文章信息/Info

Title:
Effects of MsrBl on peroxynitrite-induced cell cycle arrest in human lens epithelial cells
作者:
贾义; 钟乃凤; 周静
550025　贵州省贵安新区，贵州医科大学生物与工程学院
Author(s):
JIA Yi; ZHONG Nai-Feng; ZHOU Jing
School of Biology and Engineering , Guizhou Medical University , Gui’ an New District 550025 , Guizhou Province . China
关键词:
蛋氨酸亚砜还原酶B1; 过氧亚硝酸根; 人晶状体上皮细胞; 细胞周期; 细胞外信号调节激酶
Keywords:
methionine sulfoxide reductases Bl ; peroxynitrite ;  human lens epithelial cells ;  cell cycle ;  extracellular signal-regulated kinase
DOI:
10.13389/j.cnki.rao.2015.0279
文献标志码:
A
摘要:
目的　探讨蛋氨酸亚砜还原酶Ｂ１（ｍｅｔｈｉｏｎｉｎｅｓｕｌｆｏｘｉｄｅｒｅｄｕｃｔａｓｅＢ１，ＭｓｒＢ１）基因沉默对过氧亚硝酸根（ＯＮＯＯ－）诱导的人晶状体上皮细胞（ｈｕｍａｎｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｌｓ，ＨＬＥＣ）细胞周期的影响和ＥＲＫ信号通路在其中的调节作用。方法　将培养的ＨＬＥＣ分为两组：正常组和ＭｓｒＢ１基因沉默组，分别用０μｍｏｌ·Ｌ－１、１００μｍｏｌ·Ｌ－１、２００μｍｏｌ·Ｌ－１和３００μｍｏｌ·Ｌ－１ＯＮＯＯ－处理２０ｍｉｎ后，继续培养１２ｈ。用ＲＴ-ＰＣＲ法检测ＭｓｒＢ１基因表达水平变化，流式细胞仪检测细胞周期水平变化，Ｗｅｓｔｅｒｎｂｌｏｔ法检测ＭｓｒＢ１基因沉默和ＯＮＯＯ－对ｐＥＲＫ表达水平的影响。结果　３００μｍｏｌ·Ｌ－１ＯＮＯＯ－处理ＨＬＥＣ会导致ＭｓｒＢ１表达水平显著下降（Ｐ＜０．０１），当ＭｓｒＢ１基因沉默细胞经３００μｍｏｌ·Ｌ－１ＯＮＯＯ－处理后，ＭｓｒＢ１表达水平进一步显著下调（Ｐ＜０．０５）。经２００μｍｏｌ·Ｌ－１或３００μｍｏｌ·Ｌ－１ ＯＮＯＯ－处理，ＨＬＥＣ细胞周期分别阻滞在Ｇ２／Ｍ期和Ｓ期，当ＭｓｒＢ１基因沉默细胞经２００μｍｏｌ·Ｌ－１或３００μｍｏｌ·Ｌ－１ＯＮＯＯ－处理后，更多的细胞阻滞在Ｇ２／Ｍ期或Ｓ期（Ｐ＜０．０５）。ｐＥＲＫ检测结果表明，ＭｓｒＢ１基因沉默前后经ＯＮＯＯ－处理导致的细胞周期阻滞和ｐＥＲＫ表达水平显著下降有关（Ｐ＜０．０１）。结论　ＯＮＯＯ－可以下调ＨＬＥＣＭｓｒＢ１表达水平，ＭｓｒＢ１基因沉默细胞经ＯＮＯＯ－处理，ｐＥＲＫ表达水平显著下降，导致更多细胞阻滞在Ｇ２／Ｍ期和Ｓ期。
Abstract:
Objective To investigate the effects of methionine sulfoxide reductases Bl ( MsrBI) gene silencing on peroxynitrite-induced cell cycle arrest and the role of ERK signaling in human lens epithelial cells ( HLEC) . Methods HLEC were cultured in DMEM medium and treated with MsrBI siRNA or ONOO - (0 ymol . 1’1 ,100 ymol . 1’1 ,200 ymol . L-l and 300 ymol . L") for 12 hours. Then the effects of ONOO - on the expressing levels of MsrBI were assessed by Real-time PCR and cell cycle was quantified using flow cytometry, as well as the levels of pERK were measured by Western blot. Results The levels of MsrBI mRNA in HLEC were significantly decreased after treatment with 300 Vmol . L -l ONOO - ( P < 0. 01 ) . when MsrBI-gene-silenced cells were exposed t0 300 Vmol . L-l ONOO - . the levels of MsrBI mRNA were further decreased(P < 0. 05 ) ; The results demonstrate that treatment with 200 ymol . L-1 0r 300 ymol . L-1 0NOO - resulted in G2/M and S cell cycle arrest,respectively,when MsrBIgene-silenced cells were exposed t0 200 ymol . L-1 0r 300 ymol . L-l ONOO- .more HLEC occurred G2/M and S cell cycle arrest(P < 0. 05) . The results that cell cycle arrest in HLEC treated with ONOO - before and after MsrBlgene silencing was consistent with the results that the levels of pERK were significantly decreased ( P < 0. 01 ) . Conclusion The levels of MsrBI mRNA in HLEC are down-regulated by ONOO- .when MsrBI-gene-silenced cells are treated with ONOO - .the levels of pERK are significantly decreased . and then resulted in G2/M and S cell cycle arrest in more HLEC.

相似文献/References:

[1]贾义,宋萍萍,苏朝.过氧亚硝酸根联合碱性成纤维细胞生长因子对人晶状体上皮细胞细胞外信号调节激酶（ERK）磷酸化水平的影响[J].眼科新进展,2016,36(7):609.[doi:10.13389/j.cnki.rao.2016.0161]
　JIA Yi,SONG Ping-Ping,SU Chao.Effects of peroxynitrite and basic fibroblast growth factor on level of ERK phosphorylation in human lens epithelial cells[J].Recent Advances in Ophthalmology,2016,36(11):609.[doi:10.13389/j.cnki.rao.2016.0161]

备注/Memo

备注/Memo:
贵州省科学技术基金资助（编号：黔科合Ｊ-２０１４-２０２８）；贵阳医学院博士启动基金（编号：ＹＪ２０１４-８）
更新日期/Last Update: 2015-11-03