Deficiency in ClC-3 Chloride Channels Prevents Rat Aortic Smooth Muscle Cell Proliferation
Deficiency in ClC-3 Chloride Channels Prevents Rat Aortic Smooth Muscle Cell Proliferation
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DOI：
10.1161/01.res.0000042062.69653.e4
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发表时间：
2002-11
期刊：
影响因子：
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通讯作者：
Guan-lei Wang;Xue-Rong Wang;Mo-Jun Lin;Hua He;Xiu-jian Lan;Y. Guan
中科院分区：
文献类型：
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作者：
Guan-lei Wang;Xue-Rong Wang;Mo-Jun Lin;Hua He;Xiu-jian Lan;Y. Guan
摘要：最近越来越多的证据表明氯 (Cl−) 通道对细胞周期至关重要。在培养的大鼠主动脉血管平滑肌细胞 (VSMC) 中，我们之前发现 Cl− 通道阻滞剂可抑制内皮素-1 (ET-1) 诱导的细胞增殖。本研究旨在进一步确定负责 VSMC 增殖的特定 Cl− 通道。由于缺乏任何已知 Cl− 通道的特异性阻断剂或开放剂，我们使用反义策略来研究电压门控 Cl− 通道基因家族成员 ClC-3 在培养的大鼠主动脉 VSMC 细胞增殖中的潜在作用。通过[3H]-胸苷掺入和免疫印迹，我们发现 ET-1 诱导的细胞增殖与 ClC-3 蛋白内源表达的显着增加是平行的。用 ClC-3 特异性反义寡核苷酸瞬时转染大鼠主动脉 VSMC，以相同的浓度和时间依赖性模式抑制 ET-1 诱导的 ClC-3 蛋白表达和 VSMC 细胞增殖，而正义和错义寡核苷酸对 ClC-3 蛋白表达和细胞增殖没有影响。这些结果强烈表明 ClC-3 可能是参与 VSMC 增殖的 Cl− 通道，从而提供了将特定 Cl− 通道与细胞增殖联系起来的令人信服的分子证据。本文全文可在 http://www.circresaha.org 上获取。
Abstract— Recent growing evidence suggests that chloride (Cl−) channels are critical to the cell cycle. In cultured rat aortic vascular smooth muscle cells (VSMCs), we have previously found that Cl− channel blockers inhibit endothelin-1 (ET-1)–induced cell proliferation. The present study was designed to further identify the specific Cl− channels responsible for VSMC proliferation. Due to the lack of a specific blocker or opener of any known Cl− channels, we used the antisense strategy to investigate the potential role of ClC-3, a member of the voltage-gated Cl− channel gene family, in cell proliferation of cultured rat aortic VSMCs. With [3H]-thymidine incorporation and immunoblots, we found that ET-1–induced cell proliferation was parallel to a significant increase in the endogenous expression of ClC-3 protein. Transient transfection of rat aortic VSMCs with antisense oligonucleotide specific to ClC-3 caused an inhibition in ET-1–induced expression of ClC-3 protein and cell proliferation of VSMCs in the same concentration- and time-dependent pattern, whereas sense and missense oligonucleotides resulted in no effects on ClC-3 protein expression and cell proliferation. These results strongly suggest that ClC-3 may be the Cl− channel involved in VSMC proliferation and thus provide compelling molecular evidence linking a specific Cl− channel to cell proliferation. The full text of this article is available at http://www.circresaha.org.