Subcellular localization of aquaporin 3 in prostate cancer is regulated by RalA
Subcellular localization of aquaporin 3 in prostate cancer is regulated by RalA
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前列腺癌中水通道蛋白 3 的亚细胞定位受 RalA 调节
DOI：
10.3892/or.2018.6308
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发表时间：
2018-05-01
期刊：
影响因子：
4.2
通讯作者：
Wang, Jianbo
中科院分区：
文献类型：
--
作者：
Chen, Qiwei;Zhu, Liang;Wang, Jianbo
我们之前发现，在正常的前列腺上皮中，AQP3的定位仅限于细胞膜；而AQP3蛋白在癌上皮中的表达分布在血浆中。然而，具体机制尚不清楚。本研究构建了稳定下调RAS样原癌基因A （RalA）、过表达E-cadherin的PC-3细胞衍生物。我们发现，E-cadherin的过表达和RaLA的下调导致前列腺癌细胞质膜AQP3的升高。为了研究AQP3再分布在前列腺癌细胞中的功能，我们采用克隆性、MTT和MTS检测AQP3再分布的生长功能。采用流式细胞术和DNA阶梯TUNEL法观察其对细胞凋亡的影响。结果显示，与对照组相比，AQP3在PC-3细胞中的重分布明显抑制了细胞的增殖，增强了细胞的凋亡。伤口愈合实验和Matrigel实验表明，敲除RalA抑制了PC-3细胞的运动和侵袭能力。为了探讨AQP3在PC-3细胞中重新分布的分子机制，我们检测了PC-3细胞中cAMP的水平，结果表明，AQP3的分布受cAMP/PKA/RalA信号通路的调控。总之，这些研究提示了AQP3的新功能，并为rala导向的治疗提供了新的视角。
We previously found that in normal epithelia of the prostate, localization of AQP3 is limited to the cell membranes; however, the expression of AQP3 protein in cancer epithelia is distributed to the plasma. Yet, the detailed mechanism remains unclear. In the present study, PC-3 cell derivatives with stable knockdown of RAS like proto-oncogene A (RalA) and overexpression of E-cadherin were established. We found that overexpression of E-cadherin and knockdown of RaLA resulted in an increase in AQP3 in prostate cancer cell plasma membranes. In order to investigate the functions caused by of the AQP3 redistribution in prostate cancer cells, the growth function of AQP3 redistribution was detected with clonogenic, MTT and MTS assays. In regards to the effect on apoptosis, flow cytometric analysis and DNA Ladder TUNEL assay were utilized. The results showed that AQP3 redistribution in PC-3 cells significantly inhibited the proliferation of cells and enhanced cell apoptosis compared with these parameters in the control. Wound healing assay and Matrigel assays determined that knockout of RalA inhibited the motility and invasion capability of PC-3 cells. To investigate the molecular mechanism involved in AQP3 redistribution in PC-3 cells, the level of cAMP in PC-3 cells was examined, and the results showed that AQP3 distribution was regulated through cAMP/PKA/RalA signal pathways. In conclusion, these studies suggest a novel function of AQP3, and provide a creative view for RalA-directed therapies.