目的：探究连翘苷（Phi）通过调控Hippo/YAP信号通路对结肠癌LS180细胞增殖、迁移和侵袭的影响。方法：用不同浓度（0、5、10、20、40、80 μmol/L）的Phi处理人结肠癌LS180细胞，MTT法检测24、48 和72 h时的细胞活力。将LS180细胞分为对照组、Phi-L（5 μmol/L Phi）组、Phi-M（10 μmol/L Phi）组、Phi-H（20 μmol/L Phi）组、Phi-H+YAP抑制剂维替泊芬（VP）组（20 μmol/L Phi+5 μmol/L VP），各组均处理24 h。EdU法检测Phi对各组细胞增殖的影响，划痕愈合实验、Transwell小室法分别检测Phi对细胞迁移和侵袭的影响，免疫荧光法和WB 法检测 Phi 对细胞 Ki-67 表达率和LATS1、YAP和p-YAP、MMP-2、MMP-9、E-cadherin、N-cadherin表达的影响。构建LS180细胞移植瘤裸鼠模型，观察Phi对移植瘤体积和质量的影响，免疫荧光法和WB法检测移植瘤组织中Ki-67表达率和LATS1、YAP和p-YAP蛋白的表达水平。结果：与对照组比较，Phi-L、Phi-M和Phi-H组LS180细胞EdU阳性率、划痕愈合率、侵袭细胞数、Ki-67阳性率、MMP-2、MMP-9、N-cadherin表达均显著降低（均P<0.05），E-cadherin、LATS1和p-YAP/YAP表达均显著升高（均P<0.05）；同时使用VP则部分逆转了Phi对LS180细胞增殖、迁移与侵袭的抑制作用（均 P<0.05）Phi显著抑制裸鼠移植瘤生长，与对照组比较，Phi组裸鼠移植瘤体积、质量和 Ki-67 阳性率均显著降低（均 P<0.05），LATS1和 p-YAP/YAP 水平均显著升高（均P<0.05）。结论：Phi可能通过激活Hippo/YAP信号通路抑制结肠癌LS180细胞的恶性生物学行为。

Objective: To investigate the effects of phillyrin (Phi) on the proliferation, migration and invasion of colorectal cancer LS180 cells by regulating the Hippo/YAP signaling pathway. Methods: Human colon cancer LS180 cells were treated with Phi at different concentrations (0, 5, 10, 20, 40, 80 μmol/L), and cell vitality at 24, 48, and 72 h was detected by MTT assay. LS180 cells were divided into the Control group, the Phi-L (5 μmol/L Phi) group, the Phi-M (10 μmol/L Phi) group, the Phi-H (20 μmol/L Phi) group, and the Phi-H+YAP inhibitor verteporfin (VP) group (20 μmol/L Phi+5 μmol/L VP). EdU was applied to detect the effects of Phi on cell proliferation in each group; scratch healing test and Transwell chamber experiments were applied to detect the effects of Phi on cell migration and invasion, respectively; the effects of Phi on Ki-67 expression rate and LATS1, YAP, p-YAP, MMP-2, MMP-9, E-cadherin and N-cadherin expressions were detected by immunofluorescence and WB. A nude mouse model of LS180 cell transplanted tumor was constructed to observe the effects of Phi on the volume and mass of transplanted tumors. Ki-67 expression rate and the expression levels of LATS1, YAP and p-YAP proteins in transplanted tumor tissues were detected by immunofluorescence and WB methods. Results: Compared with the Control group, the cell EdU positive rate, scratch healing rate, cell invasion number, Ki-67 positive rate, MMP-2, MMP-9 and N-cadherin expression levels of LS180 cells in the Phi-L, Phi-M, and Phi-H groups decreased significantly (all P<0.05), while the expression levels of E-cadherin, LATS1 and p-YAP/YAP increased significantly (all P<0.05). The inhibition of Phi on the proliferation, migration and invasion of LS180 cells was partially reversed when VP was used simultaneously. Phi significantly inhibited tumor growth in nude mice. Compared with the Control group, the tumor volume, mass and Ki-67 positive rate of nude mice in the Phi group decreased significantly (all P<0.05), while the levels of LATS1 and p-YAP/YAP increased significantly (all P<0.05). Conclusion: Phi may inhibit the malignant biological behaviors of colon cancer LS180 cells by activating the Hippo/YAP signaling pathway.

济宁医学院教师科研扶持基金（No. JYFC2018FKJ078）