Ji Rong-Sheng, Wang Zi-Ling, Wu Ting, Li Zhou-Yuan, Chen Le, Deng Xin-Ran, Lu Shan, DU Hong-Zhi, Liu Da-Hui
Institute of Chinese Material Science, School of Pharmacy, Hubei University of Chinese Medicine Wuhan 430065, China.
Research Center for Pharmaceutical Preparations, Hubei University of Chinese Medicine Wuhan 430065, China.
Zhongguo Zhong Yao Za Zhi. 2023 Feb;48(3):752-761. doi: 10.19540/j.cnki.cjcmm.20221012.401.
This study explores the effect of apigenin(APG), oxymatrine(OMT), and APG+OMT on the proliferation of non-small cell lung cancer cell lines and the underlying mechanisms. Cell counting kit-8(CCK-8) assay was used to detect the vitality of A549 and NCI-H1975 cells, and colony formation assay to evaluate the colony formation ability of the cells. EdU assay was employed to examine the proliferation of NCI-H1975 cells. RT-qPCR and Western blot were performed to detect the mRNA and protein expression of PLOD2. Molecular docking was carried out to explore the direct action ability and action sites between APG/OMT and PLOD2/EGFR. Western blot was used to study the expression of related proteins in EGFR pathway. The viability of A549 and NCI-H1975 cells was inhibited by APG and APG+OMT at 20, 40, and 80 μmol·L~(-1) in a dose-dependent manner. The colony formation ability of NCI-H1975 cells was significantly suppressed by APG and APG+OMT. The mRNA and protein expression of PLOD2 was significantly inhibited by APG and APG+OMT. In addition, APG and OMT had strong binding activity with PLOD2 and EGFR. In APG and APG+OMT groups, the expression of EGFR and proteins in its downstream signaling pathways was significantly down-regulated. It is concluded that APG in combination with OMT could inhibit non-small lung cancer, and the mechanism may be related to EGFR and its downstream signaling pathways. This study lays a new theoretical basis for the clinical treatment of non-small cell lung cancer with APG in combination with OMT and provides a reference for further research on the anti-tumor mechanism of APG in combination with OMT.
本研究探讨芹菜素（APG）、氧化苦参碱（OMT）及APG+OMT对非小细胞肺癌细胞系增殖的影响及其潜在机制。采用细胞计数试剂盒-8（CCK-8）法检测A549和NCI-H1975细胞的活力，用集落形成试验评估细胞的集落形成能力。采用EdU试验检测NCI-H1975细胞的增殖情况。进行RT-qPCR和蛋白质印迹法检测PLOD2的mRNA和蛋白表达。开展分子对接以探究APG/OMT与PLOD2/EGFR之间的直接作用能力及作用位点。利用蛋白质印迹法研究EGFR通路中相关蛋白的表达。APG和APG+OMT在20、40和80μmol·L⁻¹时以剂量依赖方式抑制A549和NCI-H1975细胞的活力。APG和APG+OMT显著抑制NCI-H1975细胞的集落形成能力。APG和APG+OMT显著抑制PLOD2的mRNA和蛋白表达。此外，APG和OMT与PLOD2和EGFR具有较强的结合活性。在APG和APG+OMT组中，EGFR及其下游信号通路中蛋白的表达显著下调。结论是APG联合OMT可抑制非小细胞肺癌，其机制可能与EGFR及其下游信号通路有关。本研究为APG联合OMT临床治疗非小细胞肺癌奠定了新的理论基础，并为进一步研究APG联合OMT的抗肿瘤机制提供了参考。
