基于QCM传感器实时监测水的跨膜转运研究细胞容积调节

细胞容积调节参与细胞多种生理功能,其调节紊乱会引发多种疾病。水跨细胞膜转运是实现细胞容积快速调节的重要途径,然而,现有技术难以精确测量跨膜转运水质量动态变化表征细胞容积调节。前期研究发现,石英晶体微天平(QCM),一种纳克级质量型在线分析技术,可解决现有技术准确度差、灵敏度低、终点式测量细胞容积等难题。因此,本项目首次提出运用QCM发展新的分析平台研究细胞容积调节,构建QCM传感器实时监测水跨膜转运质量动态变化,建立高灵敏度高时间分辨测量细胞容积调节新方法；以低渗刺激发生调节性细胞容积减小(RVD)为模型,研究分别调控渗透压、容积敏感性氯通道和水通道蛋白1对癌细胞RVD功能的影响,分析几种癌细胞RVD功能差异；探讨RVD过程水跨膜转运与细胞膜通透性关系；建立可实时监测、精准测量细胞容积调节功能新方法。该研究将为细胞生理功能与水关联研究提供新的技术平台，对生理病理研究具有重要意义和应用前景。

Cell volume regulation plays an important role in various physiological processes, and its dysfunction may result in many diseases. Water transportation across plasma membrane is the main pathway for rapid cell volume regulation. However, it is infeasible to accurately detect the dynamic change of cellular mass in the process of water transportation, and to character cell volume regulation by using traditional techniques which usually suffer from poor accuracy, low sensitivity and end-point detection, etc. In our preliminary study, it is found that Quartz crystal microbalance (QCM), a kind of nanogram level of mass sensing and on-line analysis technique, can be used as an excellent tool for tracking cell volume regulation. Thus, this project firstly proposed to develop a novel analytical platform based on QCM to detect the cell volume regulation. So QCM sensor should be constructed to real-time monitoring dynamic change of cellular mass during water transportation across plasma membrane, and the new method is established for assay of cell volume regulation at high accuracy and time resolution. To evaluate the ability of cell volume regulation for different cancer cells, regulatory cell volume decrease (RVD) is used as reseach model to examine the dynamic variation in cell volume (or mass) by QCM sensor under regulating the osmotic pressure, and inhibiting of chloride channel and aquaporin 1, separately. The permeability of plasma menbrane is explored by QCM sensor monitoring water transportation and integrating with AFM and SEM imaging on nanostructure of plasma membrane. The strategy will provide a novel technique platform for relevance analysis of cell function and water, which would be applied to study physiology and pathology of diseases.