目的 研究金复康口服液对人肺腺癌PC9细胞及耐药PC9/R细胞的吉非替尼增敏作用及机制。方法 金复康口服液20 mg/mL联合不同浓度的吉非替尼（40、20、10、5、2 μmol/L）作用于PC9/R细胞，CCK-8法检测细胞增殖；培养PC9、PC9/R细胞，分为对照组、金复康口服液组、吉非替尼组、联合用药组，流式细胞术检测PC9、PC9/R细胞周期和凋亡。在BALB/c裸鼠右前肢腋窝皮下接种PC9/R细胞建立裸鼠移植瘤模型，观察金复康口服液联合吉非替尼的体内抑瘤作用；对AKT、p-AKT、PTEN、PDCD4蛋白表达的影响和对miRNA-21表达的影响。结果 与吉非替尼（2、5、10、20 μmol/L）组比较，吉非替尼（2、5、10、20 μmol/L）+金复康口服液（20 mg/mL）组的PC9/R细胞抑制率显著增加（P<0.01）；与吉非替尼组比较，联合用药可以通过显著增加早期凋亡细胞比例诱导PC9及PC9/R细胞凋亡，并使PC9、PC9/R细胞停滞在DNA合成前期，从而抑制细胞增殖，差异均具有统计学意义。体内实验表明，与吉非替尼组比较，联合用药显著抑制裸鼠PC9/R移植瘤的生长（P<0.05），且能显著降低裸鼠PC9/R组织p-AKT表达（P<0.05），增强PTEN、PDCD4的表达（P<0.05），联合用药组裸鼠PC9/R瘤组织miRNA-21的表达显著降低（P<0.05）。结论 金复康口服液能提高人肺腺癌PC9/R细胞对吉非替尼的敏感性，其作用的可能机制为通过降低miRNA-21的表达从而增强PTEN、PDCD4的表达以抑制AKT通路活性。

Objective To study investigate the mechanism of Jinfukang Oral Liquid in increasing the sensitivity of human lung adenocarcinoma PC9/R cells to gefitinib. Methods The proliferation of PC9/R cells was detected by CCK-8 method, after Jinfukang Oral Liquid 20 mg/mL combined with different concentrations of gefitinib (40, 20, 10, 5, 2 μmol/L) treating on PC9/R cells. Flow cytometry was used to detect the effects of each group on cell cycle and apoptosis of PC9 and PC9/R cells. PC9/R nude mice model was established by subcutaneous inoculation of PC9/R cells into the axilla of right forelimb of BALB/c nude mice. To observe the effect of Jinfukang oral liquid on the anti-tumor activity of gefitinib, the expression of AKT, p-AKT, PTEN and PDCD4 and the expression of microRNA-21 in nude mice xenografted with PC9/R cells. Results Compared with gefitinib (2, 5, 10, 20 μmol/L) group, the inhibition rate of PC9/R cells in gefitinib (2, 5, 10, 20 μmol/L) + Jinfukang Oral Liquid (20 mg/mL) group increased significantly (P<0.01). The combination of Jinfukang oral liquid and gefitinib could induce PC9 and PC9/R cell apoptosis by increasing the proportion of early apoptotic cells and arrest PC9 and PC9/R cells in the early stage of DNA synthesis, thus inhibiting cell proliferation. In vivo, the combination of PC9/R could inhibit the growth of PC9/R xenografts in nude mice more effectively and significantly by reducing the expression of pAKT in PC9/R tissues of nude mice (P<0.05), enhancing the expression of PTEN and PDCD4 (P<0.05). Notably, PC9/R inhibited the expression of microRNA-21 in the tumor tissues of nude mice (P<0.05). Conclusion Jinfukang Oral Liquid can enhance the sensitivity of human lung adenocarcinoma PC9/R cells to gefitinib, and its possible mechanism is to enhance the expression of PTEN and PDCD4 by reducing the expression of microRNA-21, thus inhibiting the activity ofAKT pathway.

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国家自然科学基金资助项目（81573890）；国家中医药管理局中医临床研究基地业务建设科研专题课题（JDZX2015069）；刘嘉湘国医大师传承工作室建设项目（DSGZS-2017002）；龙华医院科技创新项目（KY2017、KY1739）；上海中医药大学附属龙华医院中青年名中医培养项目