Background: Gefitinib is an important drug used to treat Non-Small Cell Lung Cancer (NSCLC) with EGFR activating mutations, but drug resistance restricts its clinical application. In this present study, combined Jin Fu Kang Decoction (JFKD) and gefitinib showed specific cytotoxicity to gefitinib-resistant cancer cells (PC-9/gef).

Objective: This study aimed to decipher the molecular mechanism of the JFKD on drug resistance when used together with Gefitinib and to find the contributing bio-active substance(s) in JFKD based on the putative mechanism.

Methods: To investigate the combined effect of gefitinib and JFKD, in vitro experiments were conducted on the established gefitinib-resistant PC-9 subclone, while in vivo experiments were conducted on the BALB/c nude mice with PC-9/gef xenografts. Western blot was used to evaluate the protein expression, and Ultra-Performance Liquid Chromatography (UPLC) coupled with quadrupole time-offlight Mass Spectrometry (MS) was used to detect the bio-active compounds of JFKD.

Results: The expression of the PTEN-relevant protein p-EGFR, p-Akt in vitro was inhibited more when combined JKFD and gefitinib were used, whereas the activities of PDCD4 and PTEN were increased; remarkably, in vivo experiments showed enhanced tumor growth inhibition when treated with this combination. Due to this combination, the effect on the gefitinib-resistant cell line, one of the JFKD-induced anti-cancer mechanisms, was found. To link the putative mechanism and the anticancer compounds in JFKD, 14 saponins and flavonoids were detected.

Conclusion: The results suggested that a promising TCM-participated therapy can be established by the putative mechanism of the combined treatment in resistant NSCLC and screening the contributing bio-active substance(s) in JFKD is meaningful on new TCM formula discovery.

背景：吉非替尼是一种用于治疗具有EGFR激活突变的非小细胞肺癌（NSCLC）的重要药物，但耐药性限制了其临床应用。在本研究中，金福康汤（JFKD）和吉非替尼联合使用对耐吉非替尼的癌细胞（PC-9 / gef）显示出特定的细胞毒性。

目的：本研究旨在探讨肯尼迪与吉非替尼一起使用时抗药性的分子机制，并在推定的机理基础上寻找肯尼迪中的重要生物活性物质。

方法：为了研究吉非替尼和JFKD的联合作用，对已建立的耐吉非替尼的PC-9亚克隆进行了体外实验，而对具有PC-9 / gef异种移植物的BALB / c裸鼠进行了体内实验。蛋白质印迹法用于评估蛋白质表达，超高效液相色谱（UPLC）结合四极杆时间-离线质谱法（MS）检测JFKD的生物活性化合物。

结果：联合JKFD和吉非替尼联合使用时，PTEN相关蛋白p-EGFR，p-Akt的体外表达受到更多抑制，而PDCD4和PTEN的活性增加。值得注意的是，体内实验显示，用这种组合物治疗后，肿瘤生长抑制作用增强。由于这种结合，发现了对吉非替尼耐药细胞系（JFKD诱导的抗癌机制之一）的作用。为了确定JFKD中的推测机制和抗癌化合物之间的联系，共检测到14种皂苷和类黄酮。

结论：结果表明，通过耐药性NSCLC联合治疗的推定机制可以建立有希望的中药参与疗法，筛选肯尼迪中的重要生物活性物质对发现新的中药配方具有重要意义。