Wei Jia, Li Yanmeng, Xu Bo, Yu Jing
Department of Oncology, Beijing Friendship Hospital, Capital Medical University No. 95 Yong An Road, Xicheng District, Beijing 100050, China.
Experimental Center, Beijing Friendship Hospital, Capital Medical University No. 95 Yong An Road, Xicheng District, Beijing 100050, China.
Am J Transl Res. 2020 May 15;12(5):1640-1657. eCollection 2020.
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have been used as first-line recommended therapy for EGFR mutant non-small cell lung cancer patients. However, epithelial-mesenchymal transition (EMT) can reduce EGFR-TKI sensitivity and lead to resistance. This study was designed to investigate the reversal effect of astragalus polysaccharides (APS) on gefitinib resistance (GR) and to elucidate the underlying mechanisms. PC9 and HCC827 lung cancer cells were stimulated by TGF-β1 to develop EMT-associated GR cells. Cell proliferation, migration and apoptosis assays were used to confirm the effect of gefitinib on GR cells and the therapeutic effect of APS on GR cells after knockdown and over-expression of related signaling pathways. Reverse transcription polymerase chain reaction, western blotting, and immunofluorescent staining assays were used to evaluate the expression levels of E-cadherin, N-cadherin, vimentin, PD-L1, and SREBP-1. Furthermore, proliferation and migration abilities were enhanced, while apoptosis ability was weakened in EMT-associated GR cells. After over-expression of PD-L1, expression levels of N-cadherin, vimentin and SREBP-1 increased, while expression of E-cadherin decreased. After knockdown of PD-L1 or SREBP-1, E-cadherin expression increased, while expression of N-cadherin and vimentin decreased. Further studies revealed that APS promoted apoptosis and reduced proliferation and migration abilities in GR cells. Moreover, APS increased expression of E-cadherin and decreased expression of N-cadherin and vimentin, indicating that it may be related to inhibition of the PD-L1/SREBP-1/EMT signaling pathway. Based on these findings, it can be concluded that APS can reverse acquired resistance to gefitinib in lung cancer cells by inhibiting the PD-L1/SREBP-1/EMT signaling pathway.
表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)已被用作表皮生长因子受体(EGFR)突变的非小细胞肺癌患者的一线推荐治疗药物。然而,上皮-间质转化(EMT)会降低EGFR-TKIs的敏感性并导致耐药。本研究旨在探讨黄芪多糖(APS)对吉非替尼耐药(GR)的逆转作用,并阐明其潜在机制。用转化生长因子-β1(TGF-β1)刺激PC9和HCC827肺癌细胞,以建立与EMT相关的GR细胞。采用细胞增殖、迁移和凋亡检测方法,确认吉非替尼对GR细胞的作用以及在相关信号通路敲低和过表达后APS对GR细胞的治疗效果。采用逆转录聚合酶链反应、蛋白质免疫印迹和免疫荧光染色检测方法,评估E-钙黏蛋白、N-钙黏蛋白、波形蛋白、程序性死亡受体配体1(PD-L1)和固醇调节元件结合蛋白1(SREBP-1)的表达水平。此外,与EMT相关的GR细胞的增殖和迁移能力增强,但凋亡能力减弱。PD-L1过表达后,N-钙黏蛋白、波形蛋白和SREBP-1的表达水平升高,而E-钙黏蛋白的表达降低。PD-L1或SREBP-1敲低后,E-钙黏蛋白的表达增加,而N-钙黏蛋白和波形蛋白的表达降低。进一步研究表明,APS可促进GR细胞凋亡,并降低其增殖和迁移能力。此外,APS增加E-钙黏蛋白的表达,降低N-钙黏蛋白和波形蛋白的表达,表明其可能与抑制PD-L1/SREBP-1/EMT信号通路有关。基于这些发现,可以得出结论:APS可通过抑制PD-L1/SREBP-1/EMT信号通路逆转肺癌细胞对吉非替尼的获得性耐药。