|
| AQP9通过调控细胞炎症及氧化应激抑制肝癌细胞转移 |
| AQP9 inhibits liver cancer cell metastasis by regulating cellular inflammation and oxidative stress |
| 投稿时间：2024-08-14 修订日期：2025-02-20 |
| DOI： |
|
 |
| 中文关键词: 肝癌 AQP9 HepG2 SMMC-7721 炎症 氧化应激 干性维持 |
| 英文关键词:Liver cancer, AQP9, HepG2, SMMC-7721, Inflammation, ROS, Stemness maintenance |
| 基金项目: |
|
| 摘要点击次数: 87 |
| 全文下载次数: 0 |
| 中文摘要: |
| 背景与目的：肝细胞癌（HCC）的转移和复发与其微环境中的炎症及氧化应激密切相关。水通道蛋白AQP9在肝癌中表达下调，但其通过调控炎症和氧化应激抑制转移的具体机制尚不明确。本研究旨在探讨AQP9是否通过调节炎症因子释放、氧化应激状态及癌细胞干性抑制肝癌转移，并解析其分子机制。方法：通过Western blot（WB）和qPCR检测不同恶性程度肝癌组织（I期、II/III期、IV期，各n=6）及癌旁正常组织中AQP9的蛋白和mRNA表达水平，结合ELISA检测促炎因子（IL-6、IL-1β、TNF-α）的表达。在低表达AQP9的HepG2和SMMC-7721细胞系中过表达AQP9，评估其对细胞迁移（划痕实验、Transwell）、增殖（克隆形成）、炎症因子（ELISA）、氧化应激指标（ROS、MDA、SOD、GSH、GSH-Px）及干性相关蛋白（CD133、Nanog、Oct4、Bmi1）的影响。通过co-IP验证AQP9调控β-catenin/TCF4/FOX3a信号通路的相互作用。结果：临床样本显示，随着肝癌恶性程度增加，AQP9在蛋白和mRNA水平显著下调（IV期较正常组织分别降至21%和22%，p<0.001），而IL-6、IL-1β、TNF-α表达升高（IV期IL-6升高3.04倍，p<0.001）。体外实验中，AQP9过表达显著抑制肝癌细胞迁移（HepG2伤口愈合率由52.2%降至29.2%，p=0.0206）和增殖（SMMC-7721克隆数减少61%，p<0.001），并降低促炎因子水平（SMMC-7721中TNF-α降至23%，p<0.001）及氧化应激标志物（ROS降低69%，MDA降低45%，p<0.01）。机制研究表明，AQP9通过增强β-catenin与FOX3a的结合，竞争性抑制β-catenin/TCF4复合体形成，从而下调干性相关基因（CD133、Nanog）。结论：AQP9通过抑制炎症反应、缓解氧化应激及调控β-catenin/TCF4/FOX3a信号通路，降低肝癌细胞干性，进而抑制其迁移和侵袭。本研究为靶向AQP9的肝癌治疗策略提供了新的理论依据。 |
| 英文摘要: |
| Background & OBJECTIVE: The metastasis and recurrence of hepatocellular carcinoma (HCC) are closely related to inflammation and oxidative stress in the microenvironment. Aquaporin AQP9 is down-regulated in liver cancer, but the specific mechanism by which AQP9 inhibits metastasis by regulating inflammation and oxidative stress remains unclear. The aim of this study was to investigate whether AQP9 can inhibit liver cancer metastasis by regulating the release of inflammatory factors, oxidative stress and dry cancer cells, and to elucidate its molecular mechanism. Methods: Western blot (WB) and qPCR were used to detect the protein and mRNA expression levels of AQP9 in different malignant HCC tissues (stage I, II/III, IV, n=6) and adjacent normal tissues, and ELISA was used to detect the expression of pro-inflammatory factors (IL-6, IL-1β, TNF-α). Overexpression of AQP9 in HepG2 and SMMC-7721 cell lines with low AQP9 expression, The effects on cell migration (scratch test, Transwell), proliferation (clonogenesis), inflammatory factors (ELISA), oxidative stress indexes (ROS, MDA, SOD, GSH, GSH-PX) and dry related proteins (CD133, Nanog, Oct4, Bmi1) were evaluated. co-IP was used to verify the interaction of AQP9 regulating β-catenin/TCF4/FOX3a signaling pathway. Results: Clinical samples showed that with the increase of malignant degree of liver cancer, the protein and mRNA levels of AQP9 were significantly down-regulated (21% and 22% in stage IV compared with normal tissues, p<0.001), while the expressions of IL-6, IL-1β and TNF-α were increased (3.04 times higher in stage IV, p<0.001). In vitro, overexpression of AQP9 significantly inhibited hepatoma cell migration (HepG2 wound healing rate decreased from 52.2% to 29.2%, p=0.0206) and proliferation (SMMC-7721 clone number decreased by 61%, p<0.001), and the level of pro-inflammatory factors decreased (TNF-α decreased to 23% in SMMC-7721, P <0.001). (p<0.001) and markers of oxidative stress (ROS decreased by 69%, MDA decreased by 45%, p<0.01). Mechanism studies have shown that AQP9 competitively inhibits the formation of β-catenin/TCF4 complex by enhancing the binding of β-catenin and FOX3a, thereby down-regulating the dry related genes (CD133, Nanog). Conclusion: AQP9 can reduce the stemness of hepatocellular carcinoma cells by inhibiting inflammatory response, relieving oxidative stress and regulating β-catenin/TCF4/FOX3a signaling pathway, and then inhibit its migration and invasion. This study provides a new theoretical basis for the treatment strategy of liver cancer targeting AQP9. |
|
在线阅读
查看/发表评论 下载PDF阅读器 |
|
|
|