[1]丁芝祥,陈阳,秦贤杰,等.miRNA-184在TGF-β2诱导人晶状体上皮细胞上皮-间质转分化中的表达[J].眼科新进展,2015,35(12):1116-1120.[doi:10.13389/j.cnki.rao.2015.0306]
　DING Zhi-Xiang,CHEN Yang,QIN Xian-Jie,et al.Expression of miR-184 in TGF-β2 induced human lens epithelial cells epithelial mesenchymal transition[J].Recent Advances in Ophthalmology,2015,35(12):1116-1120.[doi:10.13389/j.cnki.rao.2015.0306]
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miRNA-184在TGF-β2诱导人晶状体上皮细胞上皮-间质转分化中的表达
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年12期
页码:
1116-1120
栏目:
实验研究
出版日期:
2015-12-05

文章信息/Info

Title:
Expression of miR-184 in TGF-β2 induced human lens epithelial cells epithelial mesenchymal transition
作者:
丁芝祥; 陈阳; 秦贤杰; 姚微微
541001　广西壮族自治区桂林市，桂林医学院附属医院眼科
Author(s):
DING Zhi-Xiang ; CHEN Yang ;  QIN Xian-Jie ; YAO Wei-Wei
Department of Ophthalmology , Affiliated Hospital of Guilin Medical University , Guilin 541001 , Guangxi Zhuang Autonomous Region , China
关键词:
转化生长因子-β2; 人晶状体上皮细胞; miRNA-184; 上皮-间质转分化
Keywords:
transfornung growth factor β2 ;  human lens epithelial cells ;  miR-184 ;  epithelial mesenchymal transition
DOI:
10.13389/j.cnki.rao.2015.0306
文献标志码:
A
摘要:
目的　研究ｍｉＲＮＡ-１８４（ｍｉＲ-１８４）在转化生长因子β２（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒβ２，ＴＧＦ-β２）诱导人晶状体上皮细胞上皮-间质转分化过程中的表达及意义。方法　应用不同浓度（０μｇ·Ｌ－１、１μｇ·Ｌ－１、５μｇ·Ｌ－１、１０μｇ·Ｌ－１、１５μｇ·Ｌ－１）ＴＧＦ-β２ 诱导人晶状体上皮细胞发生上皮-间质转分化，细胞免疫荧光和Ｗｅｓｔｅｒｎｂｌｏｔ方法观察转分化相关分子波形蛋白（Ｖｉｍｅｎｔｉｎ）、钙黏附蛋白Ｅ（Ｅ-Ｃａｄｈｅｒｉｎ）的表达；应用ｑＲＴ-ＰＣＲ检测ｍｉＲ-１８４在不同浓度（０μｇ·Ｌ－１、１μｇ·Ｌ－１、５μｇ·Ｌ－１、１０μｇ·Ｌ－１、１５μｇ·Ｌ－１）ＴＧＦ-β２、不同作用时间（０ｈ、６ｈ、１２ｈ、２４ｈ、４８ｈ）ＴＧＦ-β２（１０μｇ·Ｌ－１）诱导人晶状体上皮细胞转分化中的表达变化。结果　随着ＴＧＦ-β２浓度的增加，细胞胞浆中Ｖｉｍｅｎｔｉｎ荧光强度增强；Ｅ-Ｃａｄｈｅｒｉｎ蛋白表达逐渐减少，各组细胞间Ｅ-Ｃａｄｈｅｒｉｎ表达量差异有统计学意义（Ｆ＝８７．６１７，Ｐ＜０．０１）；Ｖｉｍｅｎｔｉｎ蛋白表达逐渐增加，各组细胞间Ｖｉｍｅｎｔｉｎ表达量差异有统计学意义（Ｆ＝６８．９２３，Ｐ＜０．０１）。随着ＴＧＦ-β２浓度的增加，细胞中ｍｉＲ-１８４相对表达量增多，在１０μｇ·Ｌ－１ＴＧＦ-β２诱导组达到峰值；随着ＴＧＦ-β２诱导时间的延长，细胞中ｍｉＲ-１８４相对表达量增加，在２４ｈ表达达到峰值。结论　ＴＧＦ-β２诱导人晶状体上皮细胞发生上皮-间质转分化时细胞内ｍｉＲ-１８４表达增加，并与诱导浓度和时间有关，为进一步研究ｍｉＲ-１８４在人晶状体上皮细胞上皮-间质转分化中的作用及机制提供实验基础。
Abstract:
Objective To investigate the expression and sigruficance of nuR-184 in TGF-β2 induced human lens epithelial cells ( HLEC) epithelial mesenchymal transition ( EMT). Methods EMT in HLEC was induced by different concentrations of TGF-β2 (0 Vg . 1’1 ,1 Vg . 1’1 ,5 Vg . L-’,10 Vg . L-’,15 Vg . L-’) .and then immunofluorescence and Western blot were used to observe the expression of vimentin and E-Cadherin. Quantitative real-time PCR( qRT-PCR) was performed to analyze nuR-184 expressions in TGF-β2 induced HLEC at different concentration (0 yg . L-’,1 Vg . L-’, 5 Vg . 1-1 ,10 Vg . 1-1 ,15 Vg . L-’) and at different time (0 hour,6 hours , 12 hours .24 hours .48 hours) of TGF-β2 ( 10 Vg . L-’) . Results With the increase of TGF-β2 concentration.the fluorescence intensity of vimentin in cytoplasm increased. E-Cadherin protein gradually reduced .and the differences of E-Cadherin expressions among groups were statistically significant ( F = 87. 617 .P < 0. 01 ) . Meanwhile , vimentin protein gradually increased.the differences of vimentin expressions among groups were statistically sigruficant ( F = 68. 923 ,P < 0. 01 ) . With the increase of TGF- β2 concentration . the relative expression of miR-184 in HLEC increased and reached the peak in 10 yg . L-1 TGF[32 induced group. With the extension of TGF-β2 induction time , the relative expression of miR-184 in HLEC increased and reached the peak at 24 hours. Conclusion The expression of miR-184 in TGF-β2 induced EMT of HLEC is increased and related to TGF-β2 concentration and induction time ,which provides the experimental basis to further study the role and mechanism of miR-184 in EMT of HLEC.

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备注/Memo

备注/Memo:
广西自然科学基金资助（编号：２０１５ＧＸＮＳＦＡＡ１３９１７９）；桂林市科学研究与技术开发计划项目资助（编号：２０１３０１２０-４）；广西医疗卫生适宜技术研究与开发项目资助（编号：Ｓ２０１４０７-０８）
更新日期/Last Update: 2015-12-07