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研究水通道蛋白5(AQP5)对胃癌干细胞自我更新能力的作用机制及影响。

Investigating the mechanism and the effect of aquaporin 5 (AQP5) on the self-renewal capacity of gastric cancer stem cells.

作者信息

Gao Peiyao, Chen Amin, Tian Hengjin, Wang Feifan, Wang Na, Ge Kunpeng, Lian Chaoqun, Wang Fengchao, Zhang Qiang

机构信息

Department of Clinical Laboratory, The First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China.

Department of Blood Transfusion, The First Affiliated Hospital of Naval Medical University, Shanghai 200433, China.

出版信息

J Cancer. 2024 Jun 11;15(13):4313-4327. doi: 10.7150/jca.92745. eCollection 2024.

Abstract

Aquaporin 5 (AQP5) has been shown to have a pro-carcinogenic effect in numerous types of malignancies. This research intends to investigate the role and the molecular mechanism of AQP5 on enriched gastric cancer stem cells (GCSCs). : Immunohistochemistry, western blot (WB), and RT-qPCR techniques were employed to identify the presence of AQP5 in gastric cancer (GC) and the neighboring paracancerous tissues. Additionally, a statistical analysis was conducted to determine the correlation between AQP5 expression and the pathological and histological parameters. Furthermore, the study aimed to assess the predictive value of AQP5 expression in long-term survival after GC surgery. GCSCs were enriched using the serum-free culture method. The expression of AQP5 in enriched GCSCs was explored using RT-qPCR and WB. Plate cloning, transwell, WB, RT-qPCR, and the sphere-forming assay were utilized to monitor the proliferation, migration, and self-renewal capability of GCSCs after AQP5 knockdown. WB and Immunofluorescence for Detecting the Effect of AQP5 on Autophagy. WB, RT-qPCR, and other experiments were used for in-depth investigation of the potential molecular regulatory mechanism of AQP5 in GC. : AQP5 was highly expressed in GC tissues and GC cells, and overexpression of AQP5 was associated with lymph node metastasis, increased tumor size, and low 5-year postoperative survival in GC patients; other studies have shown that the AQP5 was highly expressed in GCSCs. Knockdown of AQP5 suppressed tumorigenesis and inhibited the proliferative, migratory, and self-renewal capability of GCSCs. It was also found that AQP5 could activate the autophagy phenomenon of GCSCs, and mechanistically, we found that AQP5 could regulate TRPV4 to affect the self-renewal ability of GCSCs. AQP5 can be further explored for GC therapy, as it has shown a significant impact on the self-renewal capability of GCSCs, which prevents GC progression.

摘要

水通道蛋白5(AQP5)已被证明在多种恶性肿瘤中具有促癌作用。本研究旨在探讨AQP5在富集的胃癌干细胞(GCSCs)中的作用及分子机制。采用免疫组织化学、蛋白质免疫印迹(WB)和逆转录定量聚合酶链反应(RT-qPCR)技术鉴定胃癌(GC)及癌旁组织中AQP5的存在情况。此外,进行统计分析以确定AQP5表达与病理和组织学参数之间的相关性。此外,该研究旨在评估AQP5表达对GC手术后长期生存的预测价值。采用无血清培养法富集GCSCs。运用RT-qPCR和WB技术探究富集的GCSCs中AQP5的表达情况。利用平板克隆、Transwell、WB、RT-qPCR和球体形成实验监测AQP5敲低后GCSCs的增殖、迁移和自我更新能力。采用WB和免疫荧光检测AQP5对自噬的影响。运用WB、RT-qPCR等实验深入研究AQP5在GC中的潜在分子调控机制。AQP5在GC组织和GC细胞中高表达,AQP5的过表达与GC患者的淋巴结转移、肿瘤大小增加及术后5年生存率低相关;其他研究表明AQP5在GCSCs中高表达。敲低AQP5可抑制肿瘤发生,并抑制GCSCs的增殖、迁移和自我更新能力。还发现AQP5可激活GCSCs的自噬现象,从机制上看,我们发现AQP5可调节瞬时受体电位香草酸亚型4(TRPV4)以影响GCSCs的自我更新能力。由于AQP5已显示出对GCSCs的自我更新能力有显著影响,可阻止GC进展,因此可进一步探索将其用于GC治疗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd4e/11212097/0eea92cd63fe/jcav15p4313g001.jpg

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