Kaempferol potentiates the sensitivity of pancreatic cancer cells to erlotinib via inhibition of the PI3K/AKT signaling pathway and epidermal growth factor receptor
Kaempferol potentiates the sensitivity of pancreatic cancer cells to erlotinib via inhibition of the PI3K/AKT signaling pathway and epidermal growth factor receptor
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山奈酚通过抑制 PI3K/AKT 信号通路和表皮生长因子受体增强胰腺癌细胞对厄洛替尼的敏感性
DOI：
10.1007/s10787-021-00848-1
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发表时间：
2021-07-28
影响因子：
5.8
通讯作者：
Shen, Cunsi
中科院分区：
文献类型：
--
作者：
Zhang, Zhengguang;Guo, Yuanyuan;Shen, Cunsi
厄洛替尼(erlotinib，ERL)是胰腺癌(PC)的表皮生长因子受体(EGFR)酪氨酸激酶抑制剂(TKI)。然而，由于绕过EGFR信号转导的替代通路的激活，ERL的临床疗效有限。山奈酚(KAE)是一种天然的黄酮类化合物，具有很强的抗肿瘤和抗炎作用，在本研究中，我们旨在确定KAE对PC细胞和小鼠模型ERL单一治疗的增敏作用。简要地，用CCK-8、集落形成和流式细胞术检测了KAE和ERL联合治疗对两种PC细胞系的增殖和凋亡的影响。此外，利用网络药理学方法预测了KAE和ERL的药病靶点和相关的抗PC机制。通过生存分析评估EGFR差异表达的PC患者的生存结果。分子对接技术预测了KAE与EGFR之间的亲和力。免疫印迹(WB)和免疫组织化学(IHC)分析验证相关蛋白的表达水平。结果表明，与单独使用ERL相比，KAE和ERL联合应用能显著抑制细胞增殖，促进细胞凋亡。网络药理学结果表明，KAE增敏PC对ERL的治疗可能与PI3K/AKT信号通路和EGFR TKI耐药有关。生存分析显示，EGFR高表达的PC患者生存率相对较低。分子对接结果进一步表明，KAE与 - 有很高的结合亲和力，为8.9千卡/摩尔。WB结果显示，KAE和ERL联合应用显著下调p-EGFR、p-AKT、p-ERK1/2和Bcl2的表达水平，上调caspase-9、PARP和Bax的表达水平。体内实验结果表明，KAE和ERL联合治疗进一步缩小了皮下移植瘤的体积和重量。IHC结果证实了世界卫生组织的结果。提示KAE可能通过抑制PI3K/AKT和EGFR信号通路来增强PC细胞对ERL的敏感性。
Erlotinib (ERL) is an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) of pancreatic cancer (PC). However, the clinical efficacy of ERL is limited due to the activation of alternative pathways that bypass the EGFR signaling. Kaempferol (KAE), a natural flavonoid compound, has been reported to possess potent anti-tumor and anti-inflammatory properties, and in this study, we aimed at identifying the sensitization effect of KAE on ERL monotherapy in PC cells and mouse models. Briefly, the CCK-8, colony formation, and flow cytometry were used to assess the proliferation and apoptosis of two PC cell lines in response to a treatment combination of KAE and ERL. Additionally, the drug–disease targets and related anti-PC mechanisms of KAE and ERL were predicted with a network pharmacology method. The survival outcome for PC patients with EGFR differential expression was evaluated through survival analysis. The molecular docking technique predicted the affinity between KAE and EGFR. Moreover, western blot (WB) and immunohistochemistry (IHC) analyses were applied to verify the expression levels of related proteins. As a result, in vitro results showed that the combination of KAE and ERL significantly inhibited cell proliferation and promoted cell apoptosis compared to that with ERL alone. Network pharmacology results demonstrated that KAE sensitized PC to ERL treatment may likely be related to the PI3K/AKT signaling pathway and EGFR TKI resistance. Survival analysis illustrated that PC patients with high expression of EGFR had a relative lower survival rate. Molecular docking results further suggested that KAE had a high binding affinity of – 8.9 kcal/mol with EGFR. WB results indicated that the combination of KAE and ERL dramatically downregulated the expression levels of p-EGFR, p-AKT, p-ERK1/2, and Bcl-2, and upregulated the expression levels of cleaved caspase-9, cleaved PARP, and Bax. The in vivo results revealed that treatment combination of KAE and ERL further reduced the volume and weight of subcutaneous grafted tumors. IHC results confirmed the WB results. These data imply that KAE may be a valid therapeutic candidate to potentiate PC cell sensitivity to ERL via inhibiting PI3K/AKT and EGFR signaling.