[摘要]目的：探究牛蒡子苷元（Arctigenin，ARG）对黑素瘤细胞化疗耐药性的影响以及该过程中对Hippo信号通路的调节机制。方法：利用不同浓度的维莫非尼（Vemurafenib，VEM）持续培养A375细胞以获得耐药性黑素瘤细胞株A375/VEM；CCK-8检测不同浓度VEM对A375和A375/VEM细胞的增殖活性以及不同浓度ARG对A375/VEM细胞的毒性；将A375/VEM细胞分为对照组、VEM组、ARG组、ARG+VEM组、ARG+VEM+Hippo抑制剂组，CCK-8法检测各组A375/VEM细胞的存活率；克隆形成实验检测各组A375/VEM细胞的增殖情况；流式细胞术检测各组A375/VEM细胞的凋亡情况；Western blot检测各组细胞中Cyclin D1、ERK1/2、p-ERK1/2、AKT、p-AKT及Hippo途经相关蛋白的表达；建立耐药性黑素瘤小鼠移植模型并观察肿瘤生长情况。结果：与对照组相比，VEM组、ARG组及ARG+VEM组A375/VEM细胞的存活率、克隆形成数量、Cyclin D1、p-ERK1/2、p-AKT、Yes相关蛋白（Yes-associated protein，YAP）表达均降低（P＜0.05），且ARG+VEM组低于VEM组和ARG组（P＜0.05），细胞凋亡率、LATS1、p-YAP水平升高（P＜0.05），且ARG+VEM组高于VEM组和ARG组（P＜0.05）；与ARG+VEM组相比，ARG+VEM+Hippo抑制剂组A375/VEM细胞的存活率、克隆形成数量、Cyclin D1、p-ERK1/2、p-AKT、YAP表达均升高（P＜0.05），细胞凋亡率、LATS1、p-YAP水平降低（P＜0.05）；裸鼠移植瘤体内实验发现，ARG+VEM组小鼠肿瘤重量和体积均明显低于VEM组和ARG组（P＜0.05）；进一步加入Hippo途径的抑制剂，发现ARG对A375/VEM细胞耐药性的抑制作用被逆转（P＜0.05）。结论：ARG可能是通过激活Hippo途径来抑制黑素瘤细胞的化疗耐药性。

[关键词]牛蒡子苷元（ARG）；黑素瘤细胞；化疗；耐药性；Hippo信号通路

[中图分类号]R285" " [文献标志码]A" " [文章编号]1008-6455（2025）07-0001-06

Impact of Arctigenin on Chemoresistance of Melanoma Cells by Regulating Hippo Signal Pathway

GAO Yang1， DU Ziwei1， WANG Haifei2

（ 1.Department of Dermatology， Hebei College of Traditional Chinese Medicine， Shijiazhuang 050000， Hebei， China; 2.Department of Dermatology， Luanping County Traditional Chinese Medicine Hospital， Chengde 068250， Hebei， China ）

Abstract： Objective" To explore the impact of arctigenin （ARG） on chemoresistance of melanoma cells and its regulation mechanism on Hippo signaling pathway in this process. Methods" The drug resistant melanoma cell line A375/VEM was obtained by continuously culturing A375 cells with different concentrations of vemurafenib （VEM）. CCK-8 was applied to detect the proliferative activity of VEM at different concentrations on A375 and A375/VEM cells， and the toxicity of ARG at different concentrations on A375/VEM cells. A375/VEM cells were grouped into control group， VEM group， ARG group， ARG+VEM group， and ARG+VEM+Hippo inhibitor group， CCK-8 method was applied to detect the survival rate of A375/VEM cells in each group， clone formation experiment was applied to detect the proliferation of A375/VEM cells in each group， flow cytometry was applied to detect the apoptosis of A375/VEM cells in each group. Western blot was applied to detect the expression of Cyclin D1， ERK1/2， p-ERK1/2， AKT， p-AKT， and Hippo pathway related proteins of cells in each group， the transplantation model of drug-resistant melanoma in mice was established， and the tumor growth was observed. Results" Compared with the control group， the survival rate of A375/VEM cells， clone formation quantity， the expression of Cyclin D1， p-ERK1/2， p-AKT， and Yes-associated protein in the VEM group， ARG group， and ARG+VEM group reduced （P＜0.05）， and the ARG+VEM group were lower than the VEM group and ARG group （P＜0.05）， the apoptosis rate， and the levels of LATS1 and p-YAP increased （P＜0.05）， and the ARG+VEM group were higher than the VEM group and ARG group （P＜0.05）. Compared with the ARG+VEM group， the survival rate of A375/VEM cells， clone formation quantity， the expression of Cyclin D1， p-ERK1/2， p-AKT， and YAP in the ARG+VEM+Hippo inhibitor group increased （P＜0.05）， the apoptosis rate， and the levels of LATS1 and p-YAP decreased （P＜0.05）. In vivo experiments on nude mice transplanted with tumor cells showed that the tumor weight and volume in the ARG+VEM group were obviously lower than those in the VEM and ARG groups （P＜0.05）. Further addition of Hippo pathway inhibitors revealed that the inhibitory effect of ARG on drug resistance in A375/VEM cells was reversed （P＜0.05）. Conclusion" ARG may inhibit the chemoresistance of melanoma cells by activating Hippo pathway.

Key words： arctigenin（ARG）; melanoma cells; chemotherapy; resistance; hippo signaling pathway

黑素瘤主要是黑素细胞发展而来，紫外线辐射通过其对皮肤的有害影响和直接的DNA损伤而成为皮肤黑素瘤形成的主要因素，加速肿瘤发生[1-2]。据了解，BRAF基因突变是黑素瘤最常见的突变，BRAF抑制剂维莫非尼（Vemurafenib，VEM）已被用于黑素瘤的治疗，但化疗后会产生的耐药性[3]。……