目的 基于类固醇受体共激活因子（Src）/黏着斑激酶（FAK）通路探讨淫羊藿苷对卵巢癌细胞顺铂（DDP） 耐药性的影响。方法 通过依次递增顺铂浓度的方式构建顺铂耐药 SKOV3 细胞（SKOV3/DDP）。通过 CCK-8 法检测细胞增殖率，考察淫羊藿苷及顺铂的最佳干预浓度。SKOV3/DDP 细胞分组及干预：对照组不做任何干 预；顺铂组用 5 µg·mL-1 顺铂干预 24 h；顺铂+淫羊藿苷组用 5 µg·mL-1 顺铂与 20 µg·mL-1 淫羊藿苷同时干预 24 h；顺铂+sc-3052 组用 5 µg·mL-1 顺铂与 3 µmol·L-1 sc-3052 同时干预 24 h；顺铂+淫羊藿苷+sc-3052 组用 5 µg·mL-1 顺铂、20 µg·mL-1 淫羊藿苷与 3 µmol·L-1 sc-3052 同时干预 24 h。采用 CCK-8 法检测细胞吸光度 （A450）值；克隆形成实验检测细胞增殖能力；Transwell 实验检测细胞侵袭能力；划痕实验检测细胞迁移能力； 流式细胞术检测细胞凋亡情况；Western Blot 法检测 SKOV3/DDP 细胞中多药耐药相关蛋白 5（MRP5）、多重耐 药蛋白 1（MDR1）、p-Src、p-FAK、磷酸化细胞外信号调节激酶 1/2（p-ERK1/2）蛋白表达水平。结果 （1）收集 可在 5 µg·mL-1顺铂中稳定生长的 SKOV3 细胞即为 SKOV3/DDP。选择 20 µg·mL-1淫羊藿苷、5 µg·mL-1 顺铂 作为后续实验的干预浓度。（2）与对照组比较， 顺铂组的上述各项检测指标均无明显差异（P＞0.05）。与顺铂组 比较，顺铂+淫羊藿苷组 SKOV3/DDP 细胞的 A450 值、克隆形成率、侵袭细胞数、划痕愈合率以及 MRP5、 MDR1、p-Src、p-FAK、p-ERK1/2 蛋白表达水平明显降低（P＜0.05），细胞凋亡率明显升高（P＜0.05）；顺铂 +sc-3052 组 SKOV3/DDP 细胞 A450 值、克隆形成率、侵袭细胞数、划痕愈合率以及 MRP5、MDR1、p-Src、 p-FAK、p-ERK1/2 蛋白表达水平明显升高（P＜0.05），细胞凋亡率明显降低（P＜0.05）。与顺铂+淫羊藿苷组比 较，顺铂+淫羊藿苷+sc-3052 组 SKOV3/DDP 细胞的 A450 值、克隆形成率、侵袭细胞数、划痕愈合率以及 MRP5、MDR1、p-Src、p-FAK、p-ERK1/2 蛋白表达水平明显升高（P＜0.05），细胞凋亡率明显降低（P＜ 0.05）。结论 淫羊藿苷可能通过抑制 Src/FAK 通路降低 SKOV3/DDP 细胞的顺铂耐药性。

Objective To investigate the effect of icariin on cisplatin （DDP） resistance in ovarian cancer cells based on the steroid receptor coactivator （Src）/focal adhesion kinase （FAK） pathway. Methods Cisplatin-resistant SKOV3 cells （SKOV3/DDP） were constructed by gradually increasing the concentration of cisplatin. The optimal intervention concentrations of icariin and cisplatin were determined by CCK-8 assay to measure cell proliferation rate. SKOV3/DDP cells were divided and intervened as follows：the control group received no intervention；the cisplatin group was treated with 5 µg·mL-1 cisplatin for 24 hours；the cisplatin + icariin group was treated with 5 µg·mL-1 cisplatin and 20 µg·mL-1 icariin simultaneously for 24 hours；the cisplatin + sc-3052 group was treated with 5 µg·mL-1 cisplatin and 3 µmol·L-1 sc-3052 simultaneously for 24 hours；the cisplatin + icariin + sc-3052 group was treated with 5 µg·mL-1 cisplatin， 20 µg·mL-1 icariin，and 3 µmol·L-1 sc-3052 simultaneously for 24 hours. The absorbance （A450） value of cells was detected by CCK-8 assay；cell proliferation ability was assessed by colony formation assay；cell invasion ability was evaluated by Transwell assay；cell migration ability was measured by scratch assay；cell apoptosis was detected by flow cytometry；and the protein levels of multidrug resistance-associated protein 5 （MRP5），multidrug resistance protein 1 （MDR1），p-Src，p-FAK，and phosphorylated extracellular signal-regulated kinase 1/2 （p-ERK1/2） in SKOV3/ DDP cells were determined by Western Blot. Results （1） SKOV3 cells that could stably grow in 5 µg·mL-1 cisplatin were collected as SKOV3/DDP. The concentrations of 20 µg·mL-1 icariin and 5 µg·mL-1 cisplatin were selected for subsequent experiments. （2） Compared with the control group， there were no significant differences in the above detection indicators in the cisplatin group （P＞0.05）. Compared with the cisplatin group， the A450 value， colony formation rate， number of invasive cells， scratch healing rate， and protein expression levels of MRP5， MDR1， p-Src，p-FAK，and p-ERK1/2 in the cisplatin + icariin group were significantly decreased （P＜0.05），while the apoptosis rate was significantly increased （P＜0.05）. In the cisplatin + sc-3052 group， the A450 value， colony formation rate， number of invasive cells， scratch healing rate， and protein expression levels of MRP5， MDR1， p-Src，p-FAK，and p-ERK1/2 were significantly increased （P＜0.05），while the apoptosis rate was significantly decreased （P＜0.05）. Compared with the cisplatin + icariin group，the A450 value，colony formation rate，number of invasive cells，scratch healing rate，and protein expression levels of MRP5，MDR1，p-Src，p-FAK，and p-ERK1/ 2 in the cisplatin + icariin + sc-3052 group were significantly increased （P＜0.05）， while the apoptosis rate was significantly decreased （P＜0.05）. Conclusion Icariin may reduce cisplatin resistance in SKOV3/DDP cells by inhibiting the Src/FAK pathway.

R285.5

021年度河南省医学科技攻关计划联合共建项目（LHGJ20210914）