Ion Channels and Cancer

Membrane ion channels are essential for cell proliferation and appear to have a role in the development of cancer. This has initially been demonstrated for potassium channels and is meanwhile also suggested for other cation channels and Cl⁻ channels. For some of these channels, like voltage-gated ether à go-go and Ca²⁺-dependent potassium channels as well as calcium and chloride channels, a cell cycle-dependent function has been demonstrated. Along with other membrane conductances, these channels control the membrane voltage and Ca²⁺ signaling in proliferating cells. Homeostatic parameters, such as the intracellular ion concentration, cytosolic pH and cell volume, are also governed by the activity of ion channels. Thus it will be an essential task for future studies to unravel cell cycle-specific effects of ion channels and non-specific homeostatic functions. When studying the role of ion channels in cancer cells, it is indispensable to choose experimental conditions that come close to the in vivo situation. Thus, environmental parameters, such as low oxygen pressure, acidosis and exposure to serum proteins, have to be taken into account. In order to achieve clinical application, more studies on the original cancer tissue are required, and improved animal models. Finally, it will be essential to generate more potent and specific inhibitors of ion channels to overcome the shortcomings of some of the current approaches.     

Role of Voltage-gated Potassium Channels in Cancer

Ion channels are being associated with a growing number of diseases including cancer. This overview summarizes data on voltage-gated potassium channels (VGKCs) that exhibit oncogenic properties: ether-à-go-go type 1 (Eag1). Normally, Eag1 is expressed almost exclusively in tissue of neural origin, but its ectopic expression leads to uncontrolled proliferation, while inhibition of Eag1 expression produces a concomitant reduction in proliferation. Specific monoclonal antibodies against Eag1 recognize an epitope in over 80% of human tumors of diverse origins, endowing it with diagnostic and therapeutic potential. Eag1 also possesses unique electrophysiological properties that simplify its identification. This is particularly important, as specific blockers of Eag1 currents are not available. Molecular imaging of Eag1 in live tumor models has been accomplished with dye-tagged antibodies using 3-D imaging techniques in the near-infrared spectral range. Abbreviations: EAG: Ether-à-go-go, VGKCs: voltage-gated potassium channels     

Ion Channels and Membrane Potential in Stimulus-Secretion Coupling in Adrenal Medulla Cells

Abstract: The role of Na⁺ channels and membrane potential in stimulus secretion coupling in adrenal medulla cell cultures was investigated. Veratridine, aconitine, batrachotoxin (BTX), and scorpion venom, which increase the flux of ions through tetrodotoxin(TTX)-sensitive Na⁺ channels, all evoke secretion of catecholamines that is blocked by TTX. TTX partially inhibits secretion induced by low concentrations of nicotine in Locke's solution but has no effect on high concentrations of nicotine (20 μM). In Ca²⁺-sucrose media TTX has no effect on secretion at either high or low concentrations of nicotine. Replacement of Na⁺ with Li⁺ in Locke's solution reduces the response to nicotine and to veratridine. Complete replacement of Na⁺ with hydrazine, diethanolamine, TRIS, and choline completely inhibits the response to nicotine and almost completely inhibits the response to veratridine. Following exposure of cells to 50 mM-100 mM-K⁺, nicotine does not stimulate catecholamine secretion unless the cells are resuspended in media containing less than 50 mM-K⁺. Neither dibutyryl-cyclic AMP nor dibutyryl-cyclic GMP evokes secretion. α-Bungarotoxin (1 μM) did not inhibit nicotine-induced secretion. These studies indicate that Na⁺ channels and acetylcholine (ACh) receptor ion channels are independently coupled to the influx of Ca²⁺. The membrane potential appears to affect nicotine- and veratridine-evoked secretion.     

血清分泌蛋白质组学在肿瘤中的研究进展

随着人类基因组测序计划的结束,对基因表达产物的分析已经成为研究热点,而有关蛋白质的技术亦得到了不断地发展与完善。血清分泌蛋白质组在肿瘤中起着重要作用,目前已经有不少分泌蛋白作为肿瘤标志物用于临床检测和预后判断。现对血清分泌蛋白质组在肿瘤领域的研究方法及研究进展进行了综述。     

离子通道与人类遗传病

细胞膜离子通道结构和功能正常是细胞进行生理活动的基础,对离子通道功能具有决定性意义的特定位点的突变导致其开放、关闭或激活、失活功能异常,引起组织机能紊乱,形成各种遗传性疾病。本文从水通道蛋白,钙通道,钠通道,钾通道等多种通道蛋白引起的遗传病的现象以及机理做较深入的阐述。     

Physics of Ion Channels

We review the basic physics involved in transport of ions across membrane channels in cells. Electrochemical forces that control the diffusion of ions are discussed both from microscopic and macroscopic perspectives. A case is made for use of Brownian dynamics as the minimal phenomenological model that provides a bridge between experiments and more fundamental theoretical approaches. Application of Brownian and molecular dynamics methods to channels with known molecular structures is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ion Channels Formed by NB,an Influenza B Virus Protein

The influenza B virus protein, NB, was expressed in Escherichia coli, either with a C-terminal polyhistidine tag or with NB fused to the C-terminus of glutathione S-transferase (GST), and purified by affinity chromatography. NB produced ion channel activity when added to artificial lipid bilayers separating NaCl solutions with unequal concentrations (150–500 mm cis, 50 mm trans). An antibody to a peptide mimicking the 25 residues at the C-terminal end of NB, and amantadine at high concentration (2–3 mm), both depressed ion channel activity. Ion channels had a variable conductance, the lowest conductance observed being approximately 10 picosiemens. At a pH of 5.5 to 6.5, currents reversed at positive potentials indicating that the channel was more permeable to sodium than to chloride ions (P_Na/P_Cl∼ 9). In asymmetrical NaCl solutions at a pH of 2.5, currents reversed closer to the chloride than to the sodium equilibrium potential indicating that the channel had become more permeable to chloride than to sodium ions (P_Cl/P_Na∼ 4). It was concluded that, at normal pHs, NB forms cation-selective channels. Received: 6 March 1995/Revised: 17 November 1995     

Kv1．3钾离子通道在卵巢癌细胞株中的表达及活性

目的：研究Kv1.3钾离子通道在SKOV3卵巢癌细胞中的表达及其在细胞增殖和细胞周期中的作用。方法：应用RT—PCR和免疫细胞化学鉴别Kv1.3钾离子通道在SKOV3卵巢癌细胞中的表达。应用MTT和流式细胞技术观察KV1.3钾离子通道对SKOV3卵巢癌细胞增殖及细胞周期的影响。结果：4-氨基吡啶是Kv1.3钾离子通道特异性阻滞剂。不同浓度的4－氨基吡啶可以明显抑制SKOV3细胞的增殖,并且细胞周期也受到影响。G0／G1细胞比例增加,S期和G2/M期细胞比例下降。结论：Kv1.3钾离子通道在SKOV3卵巢癌细胞中表达,并且在细胞增殖及细胞周期变换中扮演着重要的角色。     

Kv1.3 钾离子通道在卵巢癌细胞株中的表达及活性

目的:研究Kv1.3钾离子通道在SKOV3卵巢癌细胞中的表达及其在细胞增殖和细胞周期中的作用。方法:应用RT-PCR和免疫细胞化学鉴别Kv1.3钾离子通道在SKOV3卵巢癌细胞中的表达。应用MTT和流式细胞技术观察KV1.3钾离子通道对SKOV3卵巢癌细胞增殖及细胞周期的影响。结果:4-氨基吡啶是Kv1.3钾离子通道特异性阻滞剂。不同浓度的4-氨基吡啶可以明显抑制SKOV3细胞的增殖,并且细胞周期也受到影响。G0/G1细胞比例增加,S期和G2/M期细胞比例下降。结论:Kv1.3钾离子通道在SKOV3卵巢癌细胞中表达,并且在细胞增殖及细胞周期变换中扮演着重要的角色。     

离子通道与肿瘤

钾、钙、氯等离子通道在肿瘤细胞中异常表达,与肿瘤的发生发展密切相关。其可能机制是离子通道通过调节细胞膜电位、细胞周期、细胞体积、胞内钙浓度和胞质pH值等调控肿瘤细胞增殖与凋亡。本文综述了离子通道与肿瘤关系的研究进展,随着研究不断深入,离子通道有可能成为防治肿瘤的新靶标。     

Neuronal Activation Regulates the Expression of Opioid Receptors: Possible Role of Glial-Derived Factors and Voltage-Dependent Ion Channels

The previous observation that a continuous chemical depolarization of aggregating rat brain cells with KCl alters the expression of opioid receptors was examined in more detail. In contrast to its significant and converse effect on forebrain and hindbrain cells cultured in serum-containing medium, KCl had only a small and transient effect in serum-free cultures of both types. The basal receptor density in serum-free cultures was similar to the receptor density in KCl-treated serum-containing cultures, but medium conditioned by glial cells restored partially the effect of KCl in serum-free cultures. The effect of KCl in serum-containing forebrain cultures was enhanced by the voltage-dependent calcium channel blocker verapamil, and magnesium and cadmium had a similar, though smaller, effect. The sodium channel activator veratridine had a profound and dose-dependent inhibitory effect on the expression of the receptors in forebrain and hindbrain cultures, and tetrodotoxin blocked the veratridine effect. Information about the selectivity of the effect of neuronal activation on the various opioid receptor subtypes was obtained with the neuroblastoma X glioma hybrid M8 cells that possess only delta type opioid receptors. A Scatchard analysis of [3H]etorphine binding to these cells has shown that depolarization increased the Bmax, but had little, if any, effect on the affinity (KD) of the ligand to the receptors. The significance of depolarization and voltage-dependent sodium and calcium channels on the expression of different opioid receptor subtypes is discussed.     

桥粒蛋白与离子转运相关通道

桥粒为细胞与细胞之间的一种连接结构,参与细胞间机械应力传导. 在心肌组织中,桥粒与粘着连接及缝隙连接共同构成闰盘,对于维护心肌闰盘结构和功能的完整性具有重要作用. 近年来,越来越多的研究表明,桥粒蛋白基因突变、表达的缺失或功能异常,可引起心肌细胞钠、钾离子通道、缝隙连接蛋白等心肌电活动相关结构的重塑,增加心肌电学异质性,进而促发心律失常. 本文将就桥粒蛋白与离子转运相关通道关系的最新研究进展进行综述.     

锌离子对配体门控型离子通道的调节作用

在中枢神经系统(central nervous system,CNS)中,锌离子对配体门控型离子通道具有重要的调节作用。锌离子随着神经元的活动从突触前膜的囊泡中释放到突触间隙,对突触内受体进行调控。锌离子抑制N-甲基-D-天冬氨酸(N-methyl-D-aspartate,NMDA)型谷氨酸受体的活性,而对非NMDA型谷氨酸受体的调控具有多样性。由γ氨基丁酸(γ-aminobutyric acid,GABA)受体所介导的抑制性突触传递活动也受到锌离子的抑制;而锌离子对glycine受体则呈现出浓度依赖的双向调节效应。病理条件下,锌离子参与了兴奋性细胞毒作用所触发的神经元凋亡过程。本文主要阐述了在CNS中,锌离子对配体门控型离子通道所介导的突触传递活动的调控作用,以及这些调控作用的生理功能和病理意义。     

I. Ion Channels in Human THP-1 Monocytes

The THP-1 human monocytic leukemia cell line is a useful model of macrophage differentiation. Patch clamp methods were used to identify five types of ion channels in undifferentiated THP-1 monocytes. (i) Delayed rectifier K⁺ current, I _DR, was activated by depolarization to potentials positive to −50 mV, inactivated with a time constant of several hundred msec, and recovered from inactivation with a time constant ∼21 sec. I _DR was inhibited by 4-aminopyridine (4-AP), tetraethylammonium (TEA⁺), and potently by charybdotoxin (ChTX). (ii) Ca-activated K⁺ current (I _SK) dominated whole-cell currents in cells studied with 3–10 μm [Ca²⁺] _i . I _SK was at most weakly voltage-dependent, with reduced conductance at large positive potentials, and was inhibited by ChTX and weakly by TEA⁺, Cs⁺, and Ba²⁺, but not 4-AP or apamin. Block by Cs⁺ and Ba²⁺ was enhanced by hyperpolarization. (iii) Nonselective cation current, I _cat, appeared at voltages above +20 mV. Little time-dependence was observed, and a panel of channel blockers was without effect. (iv) Chloride current, I _Cl, was present early in experiments, but disappeared with time. (v) Voltage-activated H⁺ selective current is described in detail in a companion paper (DeCoursey & Cherny, 1996. J. Membrane Biol. 152:2). The ion channels in THP-1 cells are compared with channels described in other macrophage-related cells. Profound changes in ion channel expression that occur during differentiation of THP-1 cells are described in a companion paper (DeCoursey et al., 1996. J. Membrane Biol. 152:2). Received: 19 September 1995/Revised: 14 March 1996     

酸敏感离子通道的功能及其相关调控

酸敏感离子通道（ASICs）是一类由胞外酸化所激活的阳离子通道.目前,已发现了6个ASICs亚基,它们在外周和中枢神经系统中广泛表达.利用基因敲除等技术,已证明它们在触觉、痛觉、酸味觉以及学习记忆中具有重要作用.同时,它们也参与某些病理反应.ASICs可以被神经肽、温度、金属离子和缺血相关物质等调控,从而整合细胞周围的多种信号以行使其功能.     

Regulation of Sodium and Calcium Channels by Signaling Complexes

Membrane depolarization and intracellular calcium transients generated by activation of voltage-gated sodium and calcium channels are local signals, which initiate physiological processes such as action potential conduction, synaptic transmission, and excitation-contraction coupling. Targeting of effector proteins and regulatory proteins to ion channels is an important mechanism to ensure speed, specificity, and precise regulation of signaling events in response to local stimuli. In this article, we review recent experimental results showing that sodium and calcium channels form local signaling complexes, in which effector proteins, anchoring proteins, and regulatory proteins interact directly with ion channels. The intracellular domains of these channels serve as signaling platforms, mediating their participation in intracellular signaling processes. These protein-protein interactions are important for efficient synaptic transmission and for regulation of ion channels by neurotransmitters and intracellular second messengers. These localized signaling complexes are essential for normal function and regulation of electrical excitability, synaptic transmission, and excitation-contraction coupling.     

Properties of Voltage-gated Ion Channels Formed by Syringomycin E in Planar Lipid Bilayers

Using the planar lipid bilayer technique we demonstrate that the lipodepsipeptide antibiotic, syringomycin E, forms voltage-sensitive ion channels of weak anion selectivity. The formation of channels in bilayers made from dioleoylglycerophosphatidylserine doped with syringomycin E at one side (1–40 μg/ml) was greatly affected by cis-positive voltage. A change of voltage from a positive to a negative value resulted in (i) an abrupt increase in the single channel conductance (the rate of increase was voltage dependent) simultaneous with (ii) a closing of these channels and an exponential decrease in macroscopic conductance over time. The strong voltage dependence of multichannel steady state conductance, the single channel conductance, the rate of opening of channels at positive voltages and closing them at negative voltages, as well as the observed abrupt increase of single channel conductance after voltage sign reversal suggest that the change of the transmembrane field induces a significant rearrangement of syringomycin E channels, including a change in the spacing of charged groups that function as voltage sensors. The conductance induced by syringomycin E increased with the sixth power of syringomycin E concentration suggesting that at least six monomers are required for channel formation. Received: 3 April 1995/Revised: 24 August 1995