Cuan Xiangdan, Yang Xingying, Wang Jinxian, Sheng Jun, Wang Xuanjun, Huang Yanping
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural University, Kunming, China; Sanmenxia Polytechnic, Sanmenxia, China.
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural University, Kunming, China; College of Food Science and Technology, Yunnan Agricultural University, Kunming, China.
Bioorg Chem. 2024 Dec;153:107808. doi: 10.1016/j.bioorg.2024.107808. Epub 2024 Sep 7.
Epidermal growth factor receptor exon 20 insertions (EGFR Ex20ins) driver mutations in non-small cell lung cancer (NSCLC) is insensitive to EGFR tyrosine kinase inhibitors (TKIs). Therefore, it is necessary to develop more novel strategy to address the limitations of existing therapies targeting EGFR-mutated NSCLC. Lupalbigenin (LB), a flavonoid compound extracted from Derris scandens, has shown preclinical activity in lung cancer. However, the activity of LB in Ex20ins-driven tumors has not yet been elucidated. In this study, a series of stable BaF/3 cell-line that contains a high proportion (>90 %) of EGFR-eGFP Ex20ins were generated using an IL3-deprivation method. Ba/F3 cell models harboring dissimilar Ex20ins were used to characterize the antineoplastic mechanism of LB. Molecular docking confirmed that the LB could effectively bind to key target EGFR. The in vitro anticancer activity of LB was investigated in engineered Ba/F3 cells bearing diverse uncommon EGFR mutations. LB was shown to be more potent in inhibiting the viability of various uncommon EGFR-mutated cell lines. Mechanistic studies disclosed that LB repressed EGFR phosphorylation and downstream survival pathways in Ba/F3 cells expressing EGFR Ex20ins, resulting in caspase activation by activating the intrinsic apoptotic pathway. Further analyses showed that LB significantly induced G0/G1 cell cycle arrest and apoptosis in cells. LB also reduced the protein expression levels of CDK4, CDK6, CDK8, cyclin D1, cyclin A2, and Bcl2 and promoted the expression of cytochrome C, p27, and p53. In summary, we explored the possible potential targets of LB through network pharmacology and verified the target using in vitro experiments. Furthermore, our results demonstrated that LB showed potential anti-Ex20ins cancer activity through suppression of the EGFR and ERK1/2 signaling pathway in Ba/F3 cells bearing two to three amino acid insertion mutations. These findings suggested that LB might be valuable for further investigation as a potential candidate in the treatment of associated diseases.
非小细胞肺癌（NSCLC）中的表皮生长因子受体外显子20插入（EGFR Ex20ins）驱动突变对EGFR酪氨酸激酶抑制剂（TKIs）不敏感。因此，有必要开发更多新颖的策略来解决现有针对EGFR突变NSCLC疗法的局限性。鱼藤素（LB）是从毛鱼藤中提取的一种黄酮类化合物，已在肺癌中显示出临床前活性。然而，LB在Ex20ins驱动的肿瘤中的活性尚未阐明。在本研究中，使用IL3剥夺法生成了一系列稳定的BaF/3细胞系，其中含有高比例（>90%）的EGFR-eGFP Ex20ins。携带不同Ex20ins的Ba/F3细胞模型用于表征LB的抗肿瘤机制。分子对接证实LB可以有效结合关键靶点EGFR。在携带各种罕见EGFR突变的工程化Ba/F3细胞中研究了LB的体外抗癌活性。结果表明，LB在抑制各种罕见EGFR突变细胞系的活力方面更有效。机制研究表明，LB抑制表达EGFR Ex20ins的Ba/F3细胞中的EGFR磷酸化和下游存活途径，通过激活内源性凋亡途径导致半胱天冬酶激活。进一步分析表明，LB显著诱导细胞中G0/G1细胞周期停滞和凋亡。LB还降低了CDK4、CDK6、CDK8、细胞周期蛋白D1、细胞周期蛋白A2和Bcl2的蛋白表达水平，并促进了细胞色素C、p27和p53的表达。总之，我们通过网络药理学探索了LB可能的潜在靶点，并通过体外实验验证了该靶点。此外，我们的结果表明，LB通过抑制携带两到三个氨基酸插入突变的Ba/F3细胞中的EGFR和ERK1/2信号通路，显示出潜在的抗Ex20ins癌症活性。这些发现表明，LB作为治疗相关疾病的潜在候选药物可能具有进一步研究的价值。
