基于微流控芯片平台前列腺癌细胞中AQP3信号传导通路及其对肿瘤细胞侵袭能力影响的研究

Prostate carcer(PCa) is the leading cause of male cancers which account's for almost a quarter of all newly diagnosed male cancers every year. In China, more and more patients are diagnosed with PCa. PCa is a more aggressive strain and likely to metastasize than others. Water is the major component of all cells and tissues. Trans-cellular balance of this entity is the most reliable pattern of cell survival and guarantees proper cellular architecture and metabolic framework. Aquaporin3 (AQP3) functions as a selective pore allowing water, glycerol and other small solutes to pass through the plasma membrane. Presently, there's no study for going around which can descry expression and significance of AQPs in PCa. In our previous study, the expression of AQP3 mRNA and protein were examined by qRT-PCR and IHC. We found that in the normal prostatic epithelium, AQP3 localization was limited to cell membrane. However, the expression of AQP3 protein in the cancer epithelium was not observed on the cell membranes and there's some relationship between expression of AQP3 ＆ E-Cad/Cat in the PCa clinical specimens. Some studies showed that the water permeability of the plasma membrane may alter cell survival capability. It suggest that AQP3 may be involved in the sustainability and preservation of cellular character in the human PCa, which is associated with an alteration of trans-cellular water-transporting mechanisms. So, we think that changes in the localization of AQP3 in PCa cells may result from tumorigenesis. Maybe the expression of AQP3 in PCa can lead to the cell apoptotic process. May be the basolateral membrane of AQP3 can regulate not only tumor progression but also play vital part in local tissue invasion along with metastasis to distant organs through alteration of cell-to-cell contact with E-cad. In order to investigate the AQP3 biological activity, particularly in PCa progression, we designed this study. The project is divided into three parts comprising of two stages including in-vivo and in-vitro phase. We aimed to build a proper microfluidic chip to investigate the signaling pathway and the effects of AQP3 function on PCa cell apoptosis;We use Tumour Invasion Assay to study PCa cell invasion.We proceed the investigation to the next level of molecular-biological techniques for the sack of better understanding of E-Cad/Cat expression and heading further in order to dig out the logical explanation of the mechanism in PCa invasion. The core theme of our study is to find out the logical explanation of what sort of alterations do occur by the expression of AQP3 which probably lead to the apoptotic process in PCa. The second most prospective side comprises of uncovering the mystery in invasion of PCa at its molecular level which probably can lead us to a substantially promising breakthrough of new therapy protocols for treating PCa.

目前，国际上对于前列腺癌中水通道蛋白AQPs的表达及意义尚缺少研究。我们通过前期实验，发现水通道蛋白AQP3在正常前列腺上皮细胞与前列腺癌细胞中存在表达差异；AQP3在前列腺癌临床标本中的表达与E-Cad/Cat的表达有相关性；有研究表明，细胞膜上AQPs变化在细胞凋亡过程中有重要作用。故我们设想：AQP3蛋白的表达与前列腺癌细胞凋亡是否有相关性。细胞膜上AQP3向细胞质中的转移是否通过调控细胞膜上的E-Cad/Cat变化从而调控前列腺癌细胞侵袭能力？本项目拟：建立适合本实验条件的多单元集成、多单元联合、多通道连接的高通量微流控芯片实验室；在此基础上探讨AQP3表达的信号传导通路及其与前列腺癌细胞凋亡的关系；通过细胞侵袭实验，探讨AQP3的表达对于前列腺癌细胞侵袭能力的影响；探讨AQP3表达对E-Cad/Cat的影响，从而进一步探讨AQP3对前列腺癌侵袭力的调控机制。

目前，国际上对于前列腺癌中水通道蛋白AQPs的表达及意义尚缺少研究。我们通过前期实验，发现水通道蛋白AQP3在正常前列腺上皮细胞与前列腺癌细胞中存在亚细胞表达位置差异，前列腺上皮细胞主要为膜表达，而前列腺癌中AQP3主要为胞质表达；有研究表明，前列腺癌细胞中RalA与蛋白定位相关，并且与前列腺正常组织相比，RalA在前列腺癌中表达升高。故我们设想：RalA的表达与前列腺癌中AQP3位置变化是否有相关性。细胞膜上AQP3向细胞质中的转移是否调控前列腺癌细胞侵袭能力？本项目拟：建立适合本实验条件的多单元集成、多单元联合、多通道连接的高通量微流控芯片实验室；在此基础上探讨RalA表达与前列腺癌中AQP3的亚细胞位置关系，通过增殖凋亡侵袭转移实验，探讨AQP3的表达对于前列腺癌细胞生物学行为的影响；通过通路阻断剂从而进一步观察cAMP/PKA/RalA通路对AQP3的亚细胞位置影响。

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