The mechansensory Ca2+ channel in prostate tumor cells

前列腺肿瘤细胞中的机械感觉Ca2通道

• 批准号: 6919638
• 负责人: OWEN P HAMILL
• 金额: $ 12.99万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6919638
• 关键词: SDS polyacrylamide gel electrophoresis; calcium channel; cell adhesion; cell line; cell migration; confocal scanning microscopy; epithelium; fluorescence microscopy; mechanoreceptors; membrane proteins; neoplasm /cancer invasiveness; neoplastic cell; prostate neoplasms; sensory mechanism; video microscopy; voltage /patch clamp

DESCRIPTION (provided by applicant): In its early stages prostate cancer stays in the prostate and is not life-threatening, but without treatment spreads to other parts of the body and eventually causes death. A major challenge is thus to recognize and target the molecular mechanisms allowing prostate tumor cells to spread. Our long-term goal is to identify and characterize mechanosensitive and Ca2+-dependent mechanisms that regulate tumor cell migration and invasiveness. Our research is based on the emerging concept that pathophysiologically enhanced cell motility is a critical step in the metastatic cascade leading to migration and invasion by tumor cells. This project focuses on the mechanosensory Ca2+-permeable cation channels (MscCa) that transduce membrane stretch into a Ca2+ influx, thus providing feedback between cell extension and Ca2+-dependent mechanisms that generate traction forces and regulate cell adhesions. We hypothesize that specific MscCa properties acquired during tumor cell transformation allow it to coordinate migration and invasiveness. We will test this hypothesis via two specific aims comparing the functional and molecular expression of MscCa in normal human prostate epithelial and in prostate tumor cell derivatives showing transformed motility and metastatic potential vs. their parent cell lines. Aim 1 will use patch-/pressure-clamp techniques and confocal immunofluorescence microscopy to compare MscCa's properties, (including conductance, gating dynamics, membrane surface distribution, and interaction with other Ca2+-sensitive channels) between "normal" and transformed" cells. Aim 2 will use time-lapse video microscopy and fluorescent Ca2+ imaging to measure fluctuations in intracellular [Ca2+] in order to determine how MscCa and other Ca2+-sensitive channels regulate cell migration. Cell migration plays a critical role in the pathogenesis of prostate cancer, so identifying and characterizing the MscCa, a rate-limiting regulatory molecule in cell migration, should introduce an important new therapeutic target for pharmacological and genetic treatments that are urgently required to prevent the spread and mortality of prostate cancer.

描述（由申请人提供）：在早期阶段，前列腺癌停留在前列腺内，不会危及生命，但如果不进行治疗，就会扩散到身体的其他部位并最终导致死亡。因此，一个主要的挑战是识别和靶向允许前列腺肿瘤细胞扩散的分子机制。我们的长期目标是识别和表征调节肿瘤细胞迁移和侵袭的机械敏感和 Ca2+ 依赖性机制。我们的研究基于一个新兴概念，即病理生理学增强的细胞运动性是导致肿瘤细胞迁移和侵袭的转移级联中的关键步骤。该项目重点研究机械感觉 Ca2+ 渗透性阳离子通道 (MscCa)，该通道将膜拉伸转换为 Ca2+ 流入，从而在细胞延伸和产生牵引力并调节细胞粘附的 Ca2+ 依赖机制之间提供反馈。我们假设肿瘤细胞转化过程中获得的特定 MscCa 特性使其能够协调迁移和侵袭。我们将通过两个具体目标来检验这一假设，即比较正常人前列腺上皮细胞和前列腺肿瘤细胞衍生物中 MscCa 的功能和分子表达，并与其亲本细胞系相比显示出转化的运动性和转移潜力。目标 1 将使用膜片/压力钳技术和共焦免疫荧光显微镜来比较“正常”和转化的细胞之间的 MscCa 特性（包括电导、门控动力学、膜表面分布以及与其他 Ca2+ 敏感通道的相互作用）。 2 将使用延时视频显微镜和荧光 Ca2+ 成像来测量细胞内 [Ca2+] 的波动，以确定 MscCa 和其他 Ca2+ 敏感通道如何调节细胞迁移。 MscCa 在前列腺癌的发病机制中发挥着关键作用，因此鉴定和表征 MscCa（细胞迁移中的限速调节分子）应该为药物和基因治疗引入重要的新治疗靶点，而药物和基因治疗是预防前列腺癌扩散和死亡所迫切需要的。前列腺癌。