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新型机械激活离子通道 Piezo1 在人骨肉瘤细胞中的功能。

The Function of the Novel Mechanical Activated Ion Channel Piezo1 in the Human Osteosarcoma Cells.

机构信息

NingXia Medical University, Yinchuan, Ningxia, China (mainland).

Gong Li Hospital, Pudong New Area, Shanghai, China (mainland).

出版信息

Med Sci Monit. 2017 Oct 24;23:5070-5082. doi: 10.12659/msm.906959.

DOI:10.12659/msm.906959
PMID:29065102
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5665612/
Abstract

BACKGROUND The Piezo1 protein ion channel is a novel mechanical activated ion channel which is related to mechanical signal transduction. However, the function of the mechanically activated ion channel Piezo1 had not been explored. In this study, we explored the function of the Piezo1 ion channel in human osteosarcoma (OS) cells related to apoptosis, invasion, and the cell proliferation. MATERIAL AND METHODS Reverse transcription polymerase chain reaction (RT-PCR) and western-blotting were used to detect the expression of the Piezo1 protein. CCK-8, Transwell experiments and AV-PI were used to detected cell proliferation, cell invasion and cell apoptosis. RESULTS The Piezo1 protein ion channel was highly expressed in human OS cells. The Piezo1-shRNA inhibited the expression of the Piezo1. We explored whether LV3-PIEZO1-homo-3201 could act as Piezo1-shRNA, which could then be an inhibitor of Piezo1. The expression of Piezo1 in the 2-hour stretch group were slightly higher than the 0-hour stretch group, and the difference was not statistically significant (n=3, p>0.05, one-way ANOVA). The apoptotic gene such as the Bax, BAD, caspase-3, and caspase-9 had the same characteristics as the Piezo1 expression under the stretch force. We also explored the invasion of Piezo1 in vivo using nude mice, and found that Piezo1-shRNA could inhibit the invasion of the OS cells. CONCLUSIONS The Piezo1 protein may be a novel, potential therapeutic target for OS.

摘要

背景

Piezo1 蛋白离子通道是一种新型的机械激活离子通道,与机械信号转导有关。然而,Piezo1 机械激活离子通道的功能尚未被探索。在这项研究中,我们探讨了 Piezo1 离子通道在与细胞凋亡、侵袭和增殖相关的人骨肉瘤(OS)细胞中的功能。

材料和方法

采用反转录聚合酶链反应(RT-PCR)和蛋白质印迹法检测 Piezo1 蛋白的表达。用 CCK-8、Transwell 实验和 AV-PI 检测细胞增殖、细胞侵袭和细胞凋亡。

结果

Piezo1 蛋白离子通道在人骨肉瘤细胞中高表达。Piezo1-shRNA 抑制 Piezo1 的表达。我们探讨了 LV3-PIEZO1-homo-3201 是否可以作为 Piezo1-shRNA,从而成为 Piezo1 的抑制剂。在 2 小时拉伸组中 Piezo1 的表达略高于 0 小时拉伸组,但差异无统计学意义(n=3,p>0.05,单因素方差分析)。在拉伸力下,凋亡基因如 Bax、BAD、caspase-3 和 caspase-9 具有与 Piezo1 表达相同的特征。我们还在裸鼠体内探讨了 Piezo1 的侵袭,发现 Piezo1-shRNA 可以抑制 OS 细胞的侵袭。

结论

Piezo1 蛋白可能是骨肉瘤的一种新的潜在治疗靶点。

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