Id2 promotes the invasive growth of MCF-7 and SKOV-3 cells by a novel mechanism independent of dimerization to basic helix-loop-helix factors.
Id2 promotes the invasive growth of MCF-7 and SKOV-3 cells by a novel mechanism independent of dimerization to basic helix-loop-helix factors.
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Id2 通过一种独立于碱性螺旋-环-螺旋因子二聚化的新机制促进 MCF-7 和 SKOV-3 细胞的侵袭性生长
DOI：
10.1186/1471-2407-9-75
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发表时间：
2009-03-04
期刊：
影响因子：
3.8
通讯作者：
Han W
中科院分区：
文献类型：
--
作者：
Meng Y;Gu C;Wu Z;Zhao Y;Si Y;Fu X;Han W
分化抑制因子2(Id2)是脊椎动物正常发育过程中细胞增殖和分化的关键因子。Id2的大部分生物学功能都归因于其螺旋-环-螺旋基序。Id2基因在多种人类肿瘤中均有过表达，但其在肿瘤细胞侵袭潜能中的作用尚存争议。我们旨在揭示Id2在低侵袭性和雌激素受体α(ERα)阳性的MCF-7和SKOV-3癌细胞侵袭潜能中的作用。将野生型、抗降解全长或螺旋环状螺旋缺失的Id2分别稳定地导入MCF-7和SKOV-3细胞。Western印迹分析检测Id2及其突变体和E-钙粘蛋白的蛋白水平，RT-PCR检测Id2及其突变体的mRNA水平。用[3 H]-胸腺嘧啶核苷掺入法和3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四氮唑溴化法测定Id2及其突变体对细胞增殖的影响。Transwell法检测细胞体外侵袭能力。划痕实验检测细胞的运动能力。用共转染法和荧光素酶测定法检测E-钙粘蛋白启动子活性。异位转染野生型Id2可显著提高MCF-7和SKOV-3细胞中Id2蛋白和mRNA的表达，而耐降解的Id2基因可进一步提高MCF7和SKOV-3细胞中Id2的蛋白水平而不是mRNA水平。Id2或其突变体的异位表达并不改变MCF-7或SKOV-3细胞的增殖。野生型Id2能显著提高细胞的侵袭能力和迁移能力，抗降解全长Id2或HLH缺失的Id2能进一步增强野生型Id2的侵袭能力和迁移能力。E-cadherin蛋白表达和近端E-cadherin启动子的反式激活被抗降解的全长或HLH缺失的Id2显着抑制，但野生型Id2不能。E-钙粘蛋白在MCF-7和SKOV-3细胞中的异位表达仅部分钝化了耐降解的HLH缺失的Id2诱导的侵袭潜能。在ERα阳性的上皮性肿瘤细胞中过表达Id2确实通过一种不依赖于对基本螺旋-环-螺旋因子的二聚化的新机制来增加细胞的侵袭力。当Id2在某些特定细胞类型中积聚到较高水平时，E-钙粘附素仅在Id2诱导的细胞侵袭中起部分作用。
Inhibitor of differentiation 2 (Id2) is a critical factor for cell proliferation and differentiation in normal vertebrate development. Most of the biological function of Id2 has been ascribed to its helix-loop-helix motif. Overexpression of Id2 is frequently observed in various human tumors, but its role for invasion potential in tumor cells is dispute. We aimed to reveal the role of Id2 in invasion potential in poorly invasive and estrogen receptor α (ERα)-positive MCF-7 and SKOV-3 cancer cells. MCF-7 and SKOV-3 cells were stably transfected with the wild-type, degradation-resistant full-length or helix-loop-helix (HLH)-deleted Id2, respectively. Protein levels of Id2 and its mutants and E-cadherin were determined by western blot analysis and mRNA levels of Id2 and its mutants were determined by RT-PCR. The effects of Id2 and its mutants on cell proliferation were determined by [3H]-thymidine incorporation assay and the 3- [4, 5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) dye method. The in vitro invasion potential of cells was evaluated by Transwell assay. Cell motility was assessed by scratch wound assay. The promoter activity of E-cadherin was determined by cotransfection and luciferase assays. Ectopic transfection of the wild-type Id2 markedly increased the protein and mRNA expression of Id2 in MCF-7 and SKOV-3 cells; the protein level but not mRNA level was further increased by transfection with the degradation-resistant Id2 form. The ectopic expression of Id2 or its mutants did not alter proliferation of either MCF-7 or SKOV-3 cells. Transfection of the wild-type Id2 significantly induced the invasion potential and migratory capacity of cells, which was further augmented by transfection with the degradation-resistant full-length or HLH-deleted Id2. E-cadherin protein expression and transactivation of the proximal E-cadherin promoter were markedly suppressed by the degradation-resistant full-length or HLH-deleted Id2 but not wild-type Id2. Ectopic expression of E-cadherin in MCF-7 and SKOV-3 cells only partially blunted the invasion potential induced by the degradation-resistant HLH-deleted Id2. Overexpression of Id2 in ERα-positive epithelial tumor cells indeed increases the cells' invasive potential through a novel mechanism independent of dimerization to basic helix-loop-helix factors. E-cadherin contributes only in part to Id2-induced cell invasion when Id2 is accumulated to a higher level in some specific cell types.
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影响因子：
12.4
作者：
Kondo, M;Cubillo, E;Miyazono, K
通讯作者：
Miyazono, K
影响因子：
64.5
作者：
BENEZRA, R;DAVIS, RL;WEINTRAUB, H
通讯作者：
WEINTRAUB, H
影响因子：
21.3
作者：
Batlle, E;Sancho, E;de Herreros, AG
通讯作者：
de Herreros, AG
影响因子：
1.9
作者：
McLemore, Monica R.;Miaskowski, Christine;Dodd, Marylin
通讯作者：
Dodd, Marylin
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影响因子：
44.1
作者：
Meng, Yuan Guang;Han, Wei Dong;Mu, Yi Ming
通讯作者：
Mu, Yi Ming