Roles of volume-activated Cl- currents and regulatory volume decrease in the cell cycle and proliferation in nasopharyngeal carcinoma cells
Roles of volume-activated Cl- currents and regulatory volume decrease in the cell cycle and proliferation in nasopharyngeal carcinoma cells
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DOI：
10.1111/j.1365-2184.2007.00432.x
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发表时间：
2007-04-01
影响因子：
8.5
通讯作者：
Wang, L. W.
中科院分区：
文献类型：
--
作者：
Chen, L. X.;Zhu, L. Y.;Wang, L. W.
目的：之前已经表明，容量激活的质膜氯离子通道与细胞的调节性容量减少（RVD）相关，并且可能在细胞增殖的控制中发挥重要作用。我们已经证明通道的表达和 RVD 能力在细胞周期中受到积极调节。在本研究中，我们旨在进一步研究体积激活氯电流和 RVD 在细胞周期进程和细胞增殖中的作用。材料和方法：分别采用膜片钳技术、细胞图像分析技术、流式细胞术、MTT法和台盼蓝法测定或检测鼻咽癌细胞（CNE-2Z细胞）的全细胞电流、RVD、细胞周期分布、细胞增殖和细胞活力。结果：Cl-通道阻滞剂 5-硝基-2-(3-苯基丙氨基)苯甲酸 (NPPB) 和他莫昔芬以剂量依赖性方式抑制容量激活的氯电流、RVD 和 CNE-2Z 细胞的增殖。分析电流、RVD与细胞增殖的关系表明，电流、RVD均与细胞增殖呈正相关。 NPPB（100μM）和他莫昔芬（20μM）没有显着诱导细胞死亡，但抑制细胞增殖，这意味着阻断剂可能通过影响细胞周期进程来抑制细胞增殖。这通过他莫昔芬 (20 μM) 和 NPPB (100 μM) 抑制细胞周期进展并将细胞阻滞在 G(0)/G(1) 相边界的观察结果得到证实。结论：容量激活氯离子通道的活性是调节细胞通过G(1)限制点的重要因素之一，与RVD相关的Cl-电流在细胞增殖中发挥重要作用。
Objectives: Previously it has been shown, that the volume-activated plasma membrane chloride channel is associated with regulatory volume decrease (RVD) of cells and may play an important role in control of cell proliferation. We have demonstrated that both expression of the channel and RVD capacity are actively regulated in the cell cycle. In this study, we aimed to further study the role of the volume-activated chloride current and RVD in cell cycle progression and overall in cell proliferation. Materials and methods: Whole-cell currents, RVD, cell cycle distribution, cell proliferation and cell viability were measured or detected with the patch-clamp technique, the cell image analysis technique, flow cytometry, the MTT assay and the trypan blue assay respectively, in nasopharyngeal carcinoma cells (CNE-2Z cells). Results: The Cl- channel blockers, 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) and tamoxifen, inhibit the volume-activated chloride current, RVD and proliferation of CNE-2Z cells in a dose-dependent manner. Analysis of relationships between the current, RVD and cell proliferation showed that both the current and RVD were positively correlated with cell proliferation. NPPB (100 mu M) and tamoxifen (20 mu M) did not significantly induce cell death, but inhibited cell proliferation, implying that the blockers may inhibit cell proliferation by affecting cell cycle progression. This was verified by the observation that tamoxifen (20 mu M) and NPPB (100 mu M) inhibited cell cycle progress and arrested cells at the G(0)/G(1) phase boundary. Conclusions: Activity of the volume-activated chloride channel is one of the important factors that regulate the passage of cells through the G(1) restriction point and that the Cl- current associated with RVD plays an important role in cell proliferation.