RhoA和血清反应因子（SRF）介导E1A激活基因阻遏子诱导人血管平滑肌细胞分化
The Cellular Repressor of E1A-stimulated Genes Promotes Human VSMCs Differentiation In vitro Mediated by RhoA and SRF
查看参考文献12篇
|
文摘
|
为探讨E1A激活基因阻遏子（CREG）对人血管平滑肌细胞（VSMCs）分化的调控机制，应用重组逆转录病毒表达载体pLNCX2（＋）／CREG及pLXSN（-）／CREG制备稳定感染HITASY细胞模型，观察CREG蛋白过表达及表达抑制对人VSMCs分化的影响并探讨其调控机制．结果显示：pLNCX2（＋）／CREG稳定感染细胞呈分化表型，细胞细长变成组织样聚集生长趋势，细胞中CREG蛋白和平滑肌分化标志蛋白平滑肌α-肌动蛋白（SM α-actin）表达显著增加，同时SM α-aetin相关调控因子——血清反应因子（SRF）入核转位，RhoA总蛋白表达上调，以Rho激酶特异性抑制剂Y-27632作用后，CREG诱导的SM α-actin表达下调的同时SRF出核转位；pLXSN（-）／CREG稳定感染的细胞体积变大，细胞极性消失，呈无序生长，细胞中CREG和SM α-actin蛋白表达显著降低，同时伴有SRF出核转位及RhoA总蛋白表达下调．免疫共沉淀分析发现，CREG蛋白能被分泌到VSMCs培养基中表达，并可与细胞膜受体6-磷酸甘露糖／胰岛素样生长因子Ⅱ型受体（M6P／IGF2R）发生直接相互作用．用蛋白磷酸酶PP2A特异性抑制剂okadaie acid减少M6P／IGF2R在细胞膜表面分布，可明显抑制CREG过表达引起的RhoA、SRF和SM α-aetin表达．上述结果提示，在体外培养的人VSMCs中，CREG可能作为一种分泌型蛋白质通过与细胞膜受体M6P／IGF2R相互作用，依次激活SM α-actin蛋白相关调控因子RhoA和SRF引起SM α-actin表达增加，促进VSMCs向分化表型转换． |
|
其他语种文摘
|
To investigate the mechanism of the cellular repressor of E1A-stimulated genes （CREG） on the differentiation of vascular smooth muscle cells （VSMCs）, the human internal thoracic artery cells （named HITASY cells） were stably infected with sense-CREG [pLNCX2 （＋）/CREG] and antisense-CREG [pLXSN （-）/CREG] retroviral vectors and positive clone was abtained by G418 selection. The cells infected with pLNCX2（＋）/CREG take on the differentiated phenotype and overexpression of CREG can enhance SM α-actin expression accompanied with increase of the total RhoA and nuclear serum response factor （SRF） expression detected by immunofluorescence and Western blot. And treated with a selective Rho kinase inhibitor Y-27632, the expression of SM α-actin and nuclear SRF protein induced by CREG was effectively reduced. Meanwhile, the opposite results were observed in the CREG-depression HITASY cells. And immunoprecipitation assays indicate that as a secreted protein, CREG can be detected in the media of pLNCX2（＋）/CREG cells and can interact with insulin-like growth factor Ⅱ/mannose 6-phosphate receptor （M6P/IGF2R）. Furthermore, that using okadaic acid （an inhibitor of the protein phosphatases 2A） to decrease the number of M6P/IGF2R at the cell surface can significantly inhibit the expression of total RhoA, nuclear SRF and SM α-actin induced by CREG-overexpression. Taken together, as a secreted protein, CREG can enhance the expression of SM α-actin by interaction with M6P/IGF2R and perform a pivotal role in the process of VSMCs differentiation and phenotype modulation. |
|
来源
|
生物化学与生物物理进展
,2006,33(5):438-445 【核心库】
|
|
关键词
|
阻遏蛋白
;
E1A
;
表型
;
细胞
;
血管平滑肌
;
人
|
|
地址
|
1.
沈阳军区总医院全军心血管病研究所心内科, 沈阳, 110016
2.
大连市中心医院急诊科, 大连, 116033
|
|
语种
|
中文 |
|
文献类型
|
研究性论文 |
|
ISSN
|
1000-3282 |
|
学科
|
生物化学 |
|
基金
|
国家自然科学基金
|
|
文献收藏号
|
CSCD:2543256
|
参考文献 共
12
共1页
|
|
1.
韩雅玲. 对去血清后HITASY细胞分子表达及表型分析.
生物化学和生物物理进展,2003,30(6):868-873
|
CSCD被引
13
次
|
|
|
|
|
2.
韩雅玲. E1A激活基因阻遏子在不同表型血管平滑肌细胞中的表达.
中华医学杂志,2005,85(1):49-53
|
CSCD被引
9
次
|
|
|
|
|
3.
韩雅玲. E1A激活基因阻遏子过表达诱导体外培养大鼠平滑肌细胞分化.
生物化学和生物物理进展,2004,31(12):1099-1105
|
CSCD被引
10
次
|
|
|
|
|
4.
韩雅玲. 大鼠颈动脉球囊损伤后血管平滑肌细胞E1A激活基因阻遏子的表达变化.
中华心血管病杂志,2004,32(1):53-58
|
CSCD被引
15
次
|
|
|
|
|
5.
Sambrook J.
Molecular cloning:a laboratory manual.2nd,1989:1860
|
CSCD被引
1
次
|
|
|
|
|
6.
Braulke T. Role of protein phosphatases in insulin-like growth factorⅡ(IGFⅡ)-stimulated mannose 6-phosphate/IGFⅡreceptor redistribution.
J Biol Chem,1992,267(24):17347-17353
|
CSCD被引
1
次
|
|
|
|
|
7.
Rocic P. Reactive oxygen species sensitivity of angiotensinⅡ-dependent translation initiation in vascular smooth muscle cells.
J Biol Chem,2003,278(38):36973-36979
|
CSCD被引
4
次
|
|
|
|
|
8.
Liu H W. The RhoA/Rho kinase pathway regulates nuclear localization of serum response factor.
Am J Respir Cell Mol Biol,2003,29(1):39-47
|
CSCD被引
6
次
|
|
|
|
|
9.
Owens G K. Molecular regulation of vascular smooth muscle cell differentiation in development and disease.
Physiol Rev,2004,84(3):767-801
|
CSCD被引
133
次
|
|
|
|
|
10.
Veal E. The secreted glycoprotein CREG enhances differentiation of NTERA-2 human embryonal carcinoma cells.
Oncogene,2000,19(17):2120-2128
|
CSCD被引
23
次
|
|
|
|
|
11.
Di Bacco A. The secreted glycoprotein CREG inhibits cell growth dependent on the mannose-6-phosphate/insulin-like growth factorⅡreceptor.
Oncogene,2003,22(35):5436-5445
|
CSCD被引
13
次
|
|
|
|
|
12.
Gordon P V. The cellular repressor of E1A-stimulated genes mediates glucocorticoid-induced loss of the type-2 IGF receptor in ileal epithelial cells.
J Endocrinol,2005,185(2):265-273
|
CSCD被引
2
次
|
|
|
|
|
了解气象卫星更多信息，请访问