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[1]徐琳琪,齐 艳,郭如如,等.HES1在小梁网细胞中作用的初步研究[J].天津医科大学学报,2017,23(01):21-26.
　XU Lin-qi,QI Yan,GUO Ru-ru,et al.Preliminary study on the role of HES1 in human trabecular meshwork cells[J].Journal of Tianjin Medical University,2017,23(01):21-26.
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HES1在小梁网细胞中作用的初步研究(PDF)
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
23卷
期数:
2017年01期
页码:
21-26
栏目:
基础医学
出版日期:
2017-01-16

文章信息/Info

Title:
Preliminary study on the role of HES1 in human trabecular meshwork cells
文章编号:
1006-8147（2017）01-0021-06
作者:
徐琳琪 ; 齐 艳 ; 郭如如 ; 汪建涛
（天津医科大学眼科医院青光眼科，天津300384）
Author(s):
XU Lin-qi ; QI Yan; GUO Ru-ru; WANG Jian-tao
(Department of Glaucoma, Eye Hospital, Tianjin Medical University ,Tianjin 300384,China)
关键词:
HES1; 细胞外基质 ; 小梁网; 细胞迁移; 细胞增殖
Keywords:
HES1 ;  trabecular meshwork;  extracelluar matrix;  migration;  proliferation
分类号:
R775
DOI:
-
文献标志码:
A
摘要:
目的 : 利用慢病毒包装系统使人眼小梁网细胞（HTMCs）稳定表达HES1 shRNA并初步探索HES1在HTMCs的作用。 方法: 体外培养HTMCs和HEK293FT细胞并进行传代，同时应用pLKO.1-puro shRNA慢病毒载体构建HES1 shRNA质粒。Lipofectamine 2000慢病毒包装法感染原代HTMCs，使其稳定表达HES1shRNA/Scramble质粒。采用q-PCR 和western blot方法检测HES1shRNA的敲低效果及促纤维化细胞外基质蛋白（ECM）的表达变化；HTMCs增殖和迁移能力分别通过CCK-8细胞活性分析和transwell计数实验进行分析。 结果: 原代细胞经过培养后贴壁生长，呈长梭形，生长状态良好，形态符合HTMCs形态学特征。HES1 shRNA有效下调HES1 的mRNA和蛋白水平表达，P<0.05。同时，HES1 shRNA组促纤维化ECM（FN; COL1; α-SMA）的mRNA和蛋白表达水平较Scramble组显著降低，P<0.05。而Transwell计数实验显示HES1 shRNA组HTMCs的迁移数量较Scramble组增加，P<0.05；CCK-8细胞活性分析实验表明抑制HES1表达对HTMCs的增殖活性影响不大，差异不具有统计学意义，P>0.05。 结论: 成功建立了稳定表达HES1 shRNA的HTMCs细胞株。HES1可以影响HTMCs中促纤维ECM的生成，及HTMCs的增殖和迁移能力。
Abstract:
Objective: To establishhuman trabecular meshwork cells （HTMCs） cell line with stable HES1 shRNA expression and explore the role of Hairy and enhancer of split-1 (HES1) in HTMCs. Methods: Primary HTMCs and HEK293FT were cultured and passaged in vitro, and infected with HES1shRNA/Scramble lentivirus. The effects of HES1 knockdown on HES1 and profibrotic extracelluar matrix (ECM) expression were analyzed through q-PCR and Western blot. Transwell and CCK-8 counting assay were used to examine the ability of migration and proliferation HTMCs. Results: The cultured primary HTMCs were in a fibrocyte-like form with long fusiform shape and grew well. HTMCs infected with HES1 shRNA exhibited a significant decrease in HES1 and profibrotic ECM ( fibronectin , FN; collagen I , COL1; alpha-smooth muscle actin , α-SMA ) expression. HES1 shRNA treatment also increased HTMCs migration and proliferation. Conclusion: HTMCs with stable HES1 shRNA expression can be successfully built. And HES1 could affect ECM expression and celluar functions in HTMCs.

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备注/Memo

备注/Memo:
基金项目 国家自然科学基金资助项目（81270994），天津市科技计划项目基金资助（10ZCKFSY08400）

作者简介 徐琳琪（1991-），女，硕士在读，研究方向：眼视光学，青光眼方向；通信作者：汪建涛，E-mail:wangjiantao65@126.com。



更新日期/Last Update: 2017-01-16