[1]田蓉,于颖,陈有信.结缔组织生长因子在人视网膜色素上皮细胞的增生、迁移和上皮细胞-间充质细胞转变中的作用[J].眼科新进展,2014,34(12):1105-1109.[doi:10.13389/j.cnki.rao.2014.0307]
　TIAN Rong,YU Ying,CHEN You-Xin.Roles of cormective tissue growth factor in proliferation, migration and epithelial-mesenchymal transition of human retinal pigment epithelial cells[J].Recent Advances in Ophthalmology,2014,34(12):1105-1109.[doi:10.13389/j.cnki.rao.2014.0307]
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结缔组织生长因子在人视网膜色素上皮细胞的增生、迁移和上皮细胞-间充质细胞转变中的作用
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
34卷
期数:
2014年12期
页码:
1105-1109
栏目:
实验研究
出版日期:
2014-12-05

文章信息/Info

Title:
Roles of cormective tissue growth factor in proliferation, migration and epithelial-mesenchymal transition of human retinal pigment epithelial cells
作者:
田蓉; 于颖; 陈有信
100730　北京市，中国医学科学院北京协和医院眼科
Author(s):
TIAN Rong; YU Ying; CHEN You-Xin
Department of Ophthalmology,Peking Union Medical College HospitaL,Chinese Academy of Medical Science ,Beijing 100730, China
关键词:
结缔组织生长因子; 视网膜色素上皮细胞; 增生; 迁移; 上皮细胞-间充质细胞转变; 增生性玻璃体视网膜病变
Keywords:
connective tissue growth factor ;  retinal pigment epithelial cells ;  proliferation ;  nugration ;  epithelial-mesenchymal transition ;  proliferative vitreoretinopathy
DOI:
10.13389/j.cnki.rao.2014.0307
文献标志码:
A
摘要:
目的　研究结缔组织生长因子（ｃｏｎｎｅｃｔｉｖｅｔｉｓｓｕｅｇｒｏｗｔｈｆａｃｔｏｒ，ＣＴＧＦ）在人视网膜色素上皮（ｒｅｔｉｎａｌｐｉｇｍｅｎｔｅｐｉｔｈｅｌｉｕｍ，ＲＰＥ）细胞的增生、迁移和上皮细胞-间充质细胞转变（ｅｐｉｔｈｅｌｉａｌｍｅｓｅｎｃｈｙｍａｌｔｒａｎｓｉｔｉｏｎ，ＥＭＴ）中的作用。方法　采用ＣＣＫ-８细胞计数试剂盒测定４００ｎｇ?ｍＬ－１ＣＴＧＦ处理后的ＡＲＰＥ-１９细胞以及稳定表达ＣＴＧＦｓｉＲＮＡ或对照ｓｉＲＮＡ的ＡＲＰＥ-１９细胞的增生情况。采用细胞划痕实验评估ＣＴＧＦＲＮＡｉ对ＡＲＰＥ-１９细胞迁移的影响，同时测定稳定表达ＣＴＧＦｓｉＲＮＡ或对照ｓｉＲＮＡ的ＡＲＰＥ-１９细胞中ＣＴＧＦ、ＦＮ、ＭＭＰ-２和α-ＳＭＡ的表达水平以评估ＣＴＧＦ对于ＴＧＦβ１诱导的ＥＭＴ作用。结果　４００ｎｇ?ｍＬ－１ＣＴＧＦ处理组ＡＰＲＥ-１９细胞与未接受ＣＴＧＦ处理组细胞之间以及ＣＴＧＦｓｉＲＮＡ处理组与阴性对照ｓｉＲＮＡ组之间的细胞倍增时间均存在显著差异（均为Ｐ＜０．０５）。正常培养ＡＲＰＥ-１９细胞组修复划痕的时间（≤４８ｈ）显著少于ＣＴＧＦＲＮＡｉ靶向干扰处理组（≥７２ｈ）。以ＴＧＦ-β１诱导ＥＭＴ，ＣＴＧＦｓｉＲＮＡ处理组与阴性对照组比较，ＣＴＧＦ、α-ＳＭＡ、ＦＮ和ＭＭＰ-２表达均显著下调。此外，外源性ＣＴＧＦ可单独诱导ＡＲＰＥ-１９细胞α-ＳＭＡ表达增加。结论　ＣＴＧＦ能够促进ＡＲＰＥ-１９细胞增生，而ＣＴＧＦＲＮＡｉ不但能够显著抑制其增生还可显著抑制由划痕实验引发的ＡＲＰＥ-１９细胞的迁移。此外，ＣＴＧＦ可通过上调α-ＳＭＡ表达水平而增强ＡＲＰＥ-１９细胞对于ＴＧＦ-β１的反应，ＣＴＧＦＲＮＡｉ可使ＴＧＦ-β１诱导的ＥＭＴ减弱。
Abstract:
Objective To explore the roles of connective tissue growth factor ( CTGF) in the proliferation , migration and epithelial-mesenchymal transition ( EMT) of human retinal pigment epithelial ( RPE ) cells. Methods Cell proliferation of ARPE-19 cells treated with 400 ng . mL -l CTGF and ARPE-19 cells stably expressing CTGF siRNA or control siRNA were measured with a CCK-8 cell counting kit. A scratch wound healing assay was performed to evaluate the effect of CTGF RNAi on the migration of ARPE-19 cells. Meanwhile .the levels of CTGF,fibronectin ( FN) .matrix metal protease2 ( MMP-2 ) . and α-smooth muscle actin (α-SMA) in ARPE-19 cells stably expressing CTGF siRNA or control siRNA were deternuned to assess the role of CTGF in TGF-β1 induced EMT. Results There was a significant difference in the increase fold of cell numbers between ARPE-19 cells with and without CTGF treatment, and between ARPE19 cells treated with CTGF siRNA or control siRNA ( all P < 0. 05 ) . The speed of recovering the scratch in ARPE-19 control cells (《48 hours) was higher than that in cells treated with CTGF siRNA (≥ 72 hours) . Compared with ARPE-19 cells treated by control siRNA , the EMT induced by TGF-β1 was inhibited in cells treated with CTGF siRNA , represented by the significant down regulation of CTGF.α-SMA) in ARPE-19 cells stably expressing CTGF siRNA or control siRNA were deternuned —SMA. FN , and MMP-2. In addition, exogenous CTGF could induce the expression of α-SMA) in ARPE-19 cells stably expressing CTGF siRNA or control siRNA were deternuned —SMA without TGF-β1 signal in ARPE-19 cells. Conclusion CTGF can promote the proliferation of ARPE-19 cells. while CTGF RNAi significantly inhibit not only the proliferation but also the wound-healing induced migration of ARPE-19 cells. In addition , the responses of ARPE-19 cells to TGF-β1 are enhanced by CTGF by elevating the level of α-SMA) in ARPE-19 cells stably expressing CTGF siRNA or control siRNA were deternuned 一SMA,while CTGF RNAi neutralize the effects of TGF-β1 0n ARPE-19 cells to induce EMT.

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备注/Memo

备注/Memo:
国家自然科学基金面上项目基金资助（编号：３０７７２３７４）
更新日期/Last Update: 2014-12-04