缺氧诱导水通道蛋白3通过激活PI3K/Akt信号通路改变肝癌细胞对索拉非尼的敏感性。
Hypoxia-Induced Aquaporin-3 Changes Hepatocellular Carcinoma Cell Sensitivity to Sorafenib by Activating the PI3K/Akt Signaling Pathway.
作者信息
Malale Kija, Fu Jili, Qiu Liewang, Zhan Ke, Gan Xiuni, Mei Zhechuan
机构信息
Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, People's Republic of China.
Department of Endocrinology and Metabolism, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, People's Republic of China.
出版信息
Cancer Manag Res. 2020 Jun 9;12:4321-4333. doi: 10.2147/CMAR.S243918. eCollection 2020.
PURPOSE
Hypoxia-induced changes are primarily activated in patients with hepatocellular carcinoma (HCC) and long-term sorafenib exposure, thereby reducing the sensitivity to the drug. Aquaporin-3 (AQP3), a member of the aquaporin family, is a hypoxia-induced substance that affects the chemosensitivity of non-hepatocellular tumors. However, its expression and role in the sensitivity of hypoxic HCC cells to sorafenib-induced apoptosis remain unclear. The purpose of this study was to detect changes in AQP3 expression in hypoxic HCC cells and to determine whether these changes alter the sensitivity of these cells to sorafenib.
MATERIALS AND METHODS
Huh7 and HepG2 hypoxic cell models were established and AQP3 expression was detected using quantitative real-time polymerase chain reaction (qPCR) and Western blotting. Furthermore, the role of AQP3 in cell sensitivity to sorafenib was evaluated via flow cytometry, Western blotting, and a CCK-8 assay.
RESULTS
The results of qPCR and Western blotting showed that AQP3 was overexpressed in the Huh7 and HepG2 hypoxic cell models. Furthermore, AQP3 protein levels were positively correlated with hypoxia-inducible factor-1α (HIF-1α) levels. Compared with cells transfected with lentivirus-GFP (Lv-GFP), hypoxic cells transfected with lentivirus-AQP3 (Lv-AQP3) were less sensitive to sorafenib-induced apoptosis. However, the sensitivity to the drug increased in cells transfected with lentivirus-AQP3RNAi (Lv-AQP3RNAi). Akt and Erk phosphorylation was enhanced in Lv-AQP3-transfected cells. Compared with UO126 (a Mek1/2 inhibitor), LY294002 (a PI3K inhibitor) attenuated the AQP3-induced insensitivity to sorafenib observed in hypoxic cells transfected with Lv-AQP3. Combined with LY294002-treated cells, hypoxic cells transfected with Lv-AQP3RNAi were more sensitive to sorafenib.
CONCLUSION
The study results show that AQP3 is a potential therapeutic target for improving the sensitivity of hypoxic HCC cells to sorafenib.
目的
缺氧诱导的变化主要在肝细胞癌（HCC）患者和长期使用索拉非尼的情况下被激活，从而降低对该药物的敏感性。水通道蛋白3（AQP3）是水通道蛋白家族的成员，是一种缺氧诱导物质，影响非肝细胞肿瘤的化学敏感性。然而，其在缺氧HCC细胞对索拉非尼诱导凋亡的敏感性中的表达和作用仍不清楚。本研究的目的是检测缺氧HCC细胞中AQP3表达的变化，并确定这些变化是否改变这些细胞对索拉非尼的敏感性。
材料与方法
建立Huh7和HepG2缺氧细胞模型，使用定量实时聚合酶链反应（qPCR）和蛋白质印迹法检测AQP3表达。此外，通过流式细胞术、蛋白质印迹法和CCK-8试验评估AQP3在细胞对索拉非尼敏感性中的作用。
结果
qPCR和蛋白质印迹法结果显示，AQP3在Huh7和HepG2缺氧细胞模型中过表达。此外，AQP3蛋白水平与缺氧诱导因子-1α（HIF-1α）水平呈正相关。与慢病毒-GFP（Lv-GFP）转染的细胞相比，慢病毒-AQP3（Lv-AQP3）转染的缺氧细胞对索拉非尼诱导的凋亡敏感性较低。然而，慢病毒-AQP3RNAi（Lv-AQP3RNAi）转染的细胞对该药物的敏感性增加。Lv-AQP3转染的细胞中Akt和Erk磷酸化增强。与UO126（一种Mek1/2抑制剂）相比，LY294002（一种PI3K抑制剂）减弱了在Lv-AQP3转染的缺氧细胞中观察到的AQP3诱导对索拉非尼的不敏感性。与LY294002处理的细胞联合，Lv-AQP3RNAi转染的缺氧细胞对索拉非尼更敏感。
结论
研究结果表明，AQP3是提高缺氧HCC细胞对索拉非尼敏感性的潜在治疗靶点。
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