QCM sensor provides insight into the role of pivotal ions in cellular regulatory volume decrease
QCM sensor provides insight into the role of pivotal ions in cellular regulatory volume decrease
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QCM 传感器可深入了解关键离子在细胞调节体积减少中的作用
DOI：
10.1007/s00216-022-04415-7
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发表时间：
2022-11
影响因子：
4.3
通讯作者：
Xinxin Lu
中科院分区：
文献类型：
--
作者：
Peihui Yang;Shan Bao;Suting Xiao;Jingwei Feng;Xinxin Lu
所有脊椎动物细胞通常自我调节以维持体内平衡和细胞功能。作为一种主要的调节机制，调节性容积减少（RVD）发生在低渗诱导的细胞肿胀中，然后通过细胞内渗透剂和水的流出而收缩，其中K+、Cl−和Ca 2+离子在RVD过程中起关键作用。我们观察到这些关键离子在反复低渗刺激下可以导致新的RVD行为。然而，目前还缺乏有效的手段来评估关键离子对RVD的影响。在这项工作中，我们提出了一个有效的测量过程的基础上，石英晶体微天平（QCM）结合细胞功能的RVD揭示急性变化的细胞体积调节的关键离子。通过将MCF-7细胞粘附到聚赖氨酸修饰的金芯片上并用低渗NaCl介质循环刺激来实现QCM传感器，其中频率偏移（Δf）显示了该技术在表现RVD行为方面的上级可行性。随着刺激周期数的增加，在3个低渗NaCl刺激周期下，RVD值逐渐降低。与第1周期相比，第2周期和第3周期的RVD水平分别下降60.7±1.7%和82.1±1.6%（n=3），相反，在NaCl-KCl溶液中RVD水平恢复，但在NaCl-CaCl 2溶液中RVD水平显著升高52.2±0.8%。此外，抑制氯通道以阻断Cl−流出也使RVD水平降低56.2± 3.0%。结果表明，这些离子（K+、Cl-、Ca 2+）都能影响RVD的功能，其中细胞内Cl-耗尽会降低测量过程中的RVD，但补充K+后恢复，Ca 2+由于激活离子通道而增强RVD。本研究将石英晶体微天平（QCM）与细胞体积调节功能相结合，分析了关键离子在细胞中的作用，并对QCM与细胞体积调节功能之间的关系进行了探讨（K+，Cl-，Ca 2+）对调节性细胞体积减小（RVD）行为的影响。
All vertebrate cells generally self-regulate for sustaining homeostasis and cell functions. As a major regulatory mechanism, regulatory volume decrease (RVD) occurs in hypotonicity-induced cell swelling, and then shrinking by the efflux of intracellular osmolytes and water, in which the ions K+, Cl−, and Ca2+play a key role in the RVD process. We observed that these pivotal ions could result in novel RVD behaviors under repeatedly hypotonic stimulation. However, there is a lack of valid means for assessing the effect of pivotal ions on RVD. In this work, we proposed an effective measurement process based on a quartz crystal microbalance (QCM) combined with cell function of RVD for revealing acute variations in cell volume regulation induced by the pivotal ions. A QCM sensor was implemented by adhering MCF-7 cells to a poly-l-lysine-modified gold chip and cyclic stimulation with hypotonic NaCl medium, in which a frequency shift (Δf) showed the superior feasibility of the technique in exhibiting RVD behaviors. With the increase in the number of cycles, the RVD values decreased progressively under three stimulation cycles with hypotonic NaCl alone. Compared with the first cycle, the RVD level in the second and third cycles declined by 60.7±1.7% and 82.1±1.6% (n=3), respectively; conversely, it recovered in NaCl-KCl solution, but was significantly enhanced by 52.2±0.8% in NaCl-CaCl2solution. Moreover, the inhibition of chloride channels to block Cl−efflux also decreased the RVD level by 56.2±3.0%. The results indicate that these ions (K+, Cl−, Ca2+) are all able to affect the function of RVD, among which intracellular Cl−depletion reduced RVD during measurement, but which recovered with K+supplement, and Ca2+enhanced RVD due to activation of ion channels. Therefore, this work provides a comprehensive assessment of cellular behavior and offers an innovative method for gaining insight into cellular functions and mechanisms.Graphical abstractA novel strategy was conducted by integrating a quartz crystal microbalance (QCM) with the function of cell volume regulation for analyzing the role of the pivotal ions ( K+, Cl−, Ca2+) in NaCl media on the behaviors of regulatory cell volume decrease (RVD).
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影响因子：
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