PO.MCB03.03 · 分子与细胞生物学
乙酰胆碱诱导的YAP稳定化通过nAchR-alpha7/PI3K/PDPK1通路在缺氧条件下增强胰腺癌细胞的HIF-1信号
Acetylcholine-induced YAP stabilization enhances HIF-1 signaling under hypoxia via the nAchR-alpha7/PI3K/PDPK1 pathway in pancreatic cancer cells
该海报暂无可下载的资料
AACR 官方页面
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
胰腺受自主神经系统高度支配。近期研究广泛探讨了自主神经系统在胰腺导管腺癌（PDAC）发生发展中的潜在作用。缺氧诱导因子-1alpha（HIF-1alpha）通过调控与血管生成、转移和治疗抵抗相关的基因，调节肿瘤对缺氧的适应。我们此前表明，神经递质乙酰胆碱（Ach）在缺氧条件下通过nAchR-alpha7/PDPK1/YAP通路增强胰腺癌细胞中HIF-1alpha的表达，但其潜在分子机制仍不清楚。基于我们此前的发现，本研究旨在阐明Ach在缺氧条件下调控胰腺癌细胞HIF-1alpha表达的分子信号机制。在此，我们在0.5%氧气条件下用Ach处理人胰腺癌细胞。使用Western印迹、qRT-PCR和免疫共沉淀评估蛋白水平、磷酸化和相互作用。通过siRNA沉默PI3K、PDPK1和YAP的表达。在体内，使用皮下异种移植小鼠模型通过尾静脉注射Ach检验Ach的效应。我们发现，Ach激活了nAchR-alpha7下游的PI3K，增加PDPK1磷酸化。这降低了YAP在丝氨酸397位点的磷酸化，稳定了YAP并促进其核转位。在细胞核中，YAP与HIF-1alpha结合，增强其稳定性和转录活性。敲低PI3K、PDPK1或YAP可抑制Ach诱导的HIF-1alpha上调。在体内，Ach显著促进植入对照胰腺癌细胞的小鼠的肿瘤生长，但对携带nAchR-alpha7沉默细胞来源肿瘤的小鼠无影响。对临床数据集的分析显示，高nAchR-alpha7表达与PDAC患者的不良预后相关。总之，这些发现揭示Ach在缺氧条件下通过nAchR-alpha7/PI3K/PDPK1/YAP通路促进胰腺肿瘤进展，该通路稳定HIF-1alpha。这一信号轴代表了乙酰胆碱相关胰腺癌中一个有前景的治疗靶点。
查看英文原文 English abstract
The pancreas is highly innervated by the autonomic nervous system. Recent studies have extensively addressed the potential role of the autonomic nervous system in the development of pancreatic ductal adenocarcinoma (PDAC).Hypoxia-inducible factor-1alpha (HIF-1alpha) regulates tumor adaptation to hypoxia by controlling genes linked to angiogenesis, metastasis, and therapy resistance. We previously showed that neurotransmitter acetylcholine (Ach) enhances HIF-1alpha expression in pancreatic cancer cells via the nAchR-alpha7/PDPK1/YAP pathway under hypoxia, but the underlying molecular mechanisms remained unclear. Following our previous findings, this study aims to elucidate the molecular signaling mechanism by which Ach regulates HIF-1alpha expression under hypoxic conditions in pancreatic cancer cells. Here, we treated human pancreatic cancer cells with Ach under 0.5% oxygen. Protein levels, phosphorylation, and interactions were assessed using Western blot, qRT-PCR, and co-immunoprecipitation. PI3K, PDPK1, and YAP expression were silenced by siRNA. In vivo, the effect of Ach was tested using tail-vein Ach injections in a subcutaneous xenograft mouse model. We found that Ach activated PI3K downstream of nAchR-alpha7, increasing PDPK1 phosphorylation. This reduced YAP phosphorylation at serine 397, stabilizing YAP and promoting its nuclear translocation. In the nucleus, YAP bound to HIF-1alpha, enhancing its stability and transcriptional activity. Knockdown of PI3K, PDPK1, or YAP suppressed Ach-induced HIF-1alpha upregulation. In vivo , Ach significantly promoted tumor growth in mice implanted with control pancreatic cancer cells, but had no effect in mice bearing tumors derived from nAchR-alpha7-silenced cells. Analysis of clinical datasets revealed that high nAchR-alpha7 expression correlates with poor prognosis in PDAC patients. Collectively, these findings reveal that Ach promotes pancreatic tumor progression under hypoxia via the nAchR-alpha7/PI3K/PDPK1/YAP pathway, which stabilizes HIF-1alpha. This signaling axis represents a promising therapeutic target in acetylcholine-associated pancreatic cancer.
利益披露 Disclosure
Y. Cho, None..
H. Kim, None..
E. Oh, None.