Regulation of cell proliferation by intermediate-conductance Ca2+-activated potassium and volume-sensitive chloride channels in mouse mesenchymal stem cells
Regulation of cell proliferation by intermediate-conductance Ca2+-activated potassium and volume-sensitive chloride channels in mouse mesenchymal stem cells
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DOI：
10.1152/ajpcell.00268.2008
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发表时间：
2008-11-01
影响因子：
5.5
通讯作者：
Li, Gui-Rong
中科院分区：
文献类型：
--
作者：
Tao, Rong;Lau, Chu-Pak;Li, Gui-Rong
Tao R，Lau CP，Tse HF，Li GR.小鼠间充质干细胞中中钙激活钾和体积敏感性氯离子通道对细胞增殖的调节。美国生理学杂志细胞生理学295：C1409 C1416，2008年。首次发表于2008年9月24日; doi：10.1152/ajpcell.00268.2008。骨髓间充质干细胞（MSC）是再生医学的一种有前途的细胞来源;然而，它们的细胞生理学尚未完全了解。本研究旨在探讨两种主要的功能性离子通道--中电导钙激活钾通道（IKCa）和体积敏感氯通道（I-Cl.vol）在调控小鼠骨髓间充质干细胞增殖中的潜在作用。我们发现，抑制IKCa与克霉唑和I-Cl.vol与5-硝基-1-（3-苯基丙基氨基）苯甲酸（NPPB）以浓度依赖性的方式减少细胞增殖。用特异性短干扰（si）RNA敲低KCa3.1或Clcn 3显著降低IKCa或ICl.vol密度和通道蛋白，并产生细胞增殖的显著抑制（分别为24.4 +/- 9.6%和29.5 +/-7.2%，与对照相比P < 0.05）。流式细胞仪分析显示，小鼠骨髓间充质干细胞仍停留在G（0）/G（1）期（对照：51.65 +/- 3.43%），通过使用克霉唑抑制IKCa或I-Cl.vol（2 μ M：64.45 +/-2.20%，P < 0.05）或NPPB（200 μ M：82.89 +/-2.49%，P < 0.05）或特异性siRNA，同时S期细胞分布减少。Western blot分析显示细胞周期调控蛋白cyclin D1和cyclin E的表达减少。总的来说，我们的研究结果表明，IKCa和I-Cl.vol通道通过调节细胞周期蛋白D1和细胞周期蛋白E的表达来调节细胞周期进程和小鼠MSC的增殖。
Tao R, Lau CP, Tse HF, Li GR. Regulation of cell proliferation by intermediate-conductance Ca2+-activated potassium and volume-sensitive chloride channels in mouse mesenchymal stem cells. Am J Physiol Cell Physiol 295: C1409C1416, 2008. First published September 24, 2008; doi: 10.1152/ajpcell.00268.2008.-Bone marrow mesenchymal stem cells (MSCs) are a promising cell source for regenerative medicine; however, their cellular physiology is not fully understood. The present study aimed at exploring the potential roles of the two dominant functional ion channels, intermediate-conductance Ca2+-activated potassium (IKCa) and volume-sensitive chloride (I-Cl.vol) channels, in regulating proliferation of mouse MSCs. We found that inhibition of IKCa with clotrimazole and I-Cl.vol with 5-nitro-1-(3-phenylpropylamino) benzoic acid (NPPB) reduced cell proliferation in a concentration-dependent manner. Knockdown of KCa3.1 or Clcn3 with specific short interference (si) RNAs significantly reduced IKCa or ICl.vol density and channel protein and produced a remarkable suppression of cell proliferation (by 24.4 +/- 9.6% and 29.5 +/- 7.2%, respectively, P < 0.05 vs. controls). Flow cytometry analysis showed that mouse MSCs retained at G(0)/G(1) phase (control: 51.65 +/- 3.43%) by inhibiting IKCa or I-Cl.vol using clotrimazole (2 mu M: 64.45 +/- 2.20%, P < 0.05) or NPPB (200 mu M: 82.89 +/- 2.49%, P < 0.05) or the specific siRNAs, meanwhile distribution of cells in S phase was decreased. Western blot analysis revealed a reduced expression of the cell cycle regulatory proteins cyclin D1 and cyclin E. Collectively, our results have demonstrated that IKCa and I-Cl.vol channels regulate cell cycle progression and proliferation of mouse MSCs by modulating cyclin D1 and cyclin E expression.