Proliferation of human lens epithelial cells (HLE-B3) is inhibited by blocking of voltage-gated calcium channels
Proliferation of human lens epithelial cells (HLE-B3) is inhibited by blocking of voltage-gated calcium channels
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DOI：
10.1007/s00424-008-0514-5
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发表时间：
2008-05
期刊：
影响因子：
--
通讯作者：
A. Meissner;T. Noack
中科院分区：
文献类型：
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作者：
A. Meissner;T. Noack
钙离子作为众多细胞调控途径的组成部分，具有选择性地控制细胞周期中特定的细胞功能，如G1期的开始。细胞增殖依赖于细胞内钙离子水平的增加。细胞内钙的主要调节途径是钙通过电压门控钙通道流入细胞。人透镜上皮细胞（hLEC）中存在大量T型和L型钙通道，米贝拉地尔可抑制总钙电流。在这里，如果钙内流通过Cavchannels调节上皮细胞的增殖，假设进行了测试。细胞增殖通过细胞培养试验测定，使用L-和T-型Cavchannel阻断剂米贝拉地尔和维拉帕米作为钙内流的调节剂。使用锰淬火技术研究钙内流。蛋白质印迹实验在标准条件下使用针对MAPK 3的抗体完成。米贝拉地尔以及维拉帕米损害细胞增殖，但在不同的浓度范围。此外，两种拮抗剂均降低了MAPK 3的激活。在两种阻滞剂存在下，钙内流也减少。我们的结论是Ca 2+通过Cavchannels的跨膜内流有助于调节hLEC的增殖，确定Cavchannel阻滞剂作为潜在的治疗物质在眼部疾病。
Calcium, as an integral part of a large number of cellular regulatory pathways, is selective in the control of specific cell functions like the start of G1 phase in cell cycle. Cell proliferation has been suggested to depend on increasing intracellular calcium levels. A major regulatory pathway for intracellular calcium is the calcium influx into the cell via voltage-gated calcium channels. T-type and L-type calcium channels are substantially present in human lens epithelial cell (hLEC), and total calcium currents are inhibited by mibefradil. Here, the hypothesis was tested if calcium influx via Cavchannels regulates proliferation in epithelial cells. Cell proliferation was determined by cell culture assays using the L- and T-type Cavchannel blockers mibefradil and verapamil as modulators for calcium influx. Calcium influx was investigated using the Manganese quench technique. Western blot experiments were accomplished under standard conditions using antibodies against MAPK 3. Mibefradil as well as verapamil impaired cell proliferation, but in different concentration ranges. Furthermore, the activation of MAPK 3 was reduced by both antagonists. Calcium influx was also reduced in the presence of both blockers. We conclude that the transmembrane influx of Ca2+through Cavchannels contributes to the regulation of hLEC proliferation, identifying Cavchannel blockers as potential therapeutic substances in ocular diseases.