水通道蛋白5(AQP5)已被证明在许多类型的恶性肿瘤中具有促致癌作用。本研究旨在探讨AQP5对富集型胃癌干细胞(GCSCs)的作用及其分子机制。方法:免疫组织化学,蛋白质印迹(WB),采用RT-qPCR技术鉴定胃癌(GC)及其癌旁组织中AQP5的存在。此外,进行统计学分析以确定AQP5表达与病理和组织学参数之间的相关性.此外,本研究旨在评估AQP5表达对GC术后长期生存的预测价值.使用无血清培养方法富集GCSC。使用RT-qPCR和WB探索富集的GCSCs中AQP5的表达。平板克隆,transwell,WB,RT-qPCR,并利用球体形成试验来监测增殖,迁移,AQP5敲除后GCSCs的自我更新能力。WB和免疫荧光检测AQP5对自噬的影响.WB,RT-qPCR,等实验用于深入研究AQP5在GC中的潜在分子调控机制。结果:AQP5在GC组织和GC细胞中高表达,AQP5的过表达与淋巴结转移有关,肿瘤大小增加,GC患者术后5年生存率较低;其他研究表明AQP5在GCSCs中高表达。敲除AQP5抑制体内肿瘤发生并抑制增殖,迁徙,和GCSCs的自我更新能力。还发现AQP5可以激活GCSCs的自噬现象,在机械上,我们发现AQP5可以调节TRPV4从而影响GCSCs的自我更新能力。结论:AQP5可进一步探索用于GC治疗,因为它对GCSC的自我更新能力有重大影响,这阻止了GC进展。
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). Methods: 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. Results:
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 in vivo 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. Conclusion: 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.