[乐伐替尼在治疗瑞戈非尼耐药的肝细胞癌中下调IGF1R/Mek/Erk信号通路]

[Lenvatinib down-regulates IGF1R/Mek/Erk signaling pathway in the treatment of regorafenib-resistant hepatocellular carcinoma].

作者信息

Zhang J, Kuang S W, Huang N, Zhang J J, Liu M, Wang L M

机构信息

Department of Hepatobiliary Surgery, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.

Department of Oncology, Xiangya Hospital, Central South University, Changsha 410008, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2023 Jun 23;45(6):490-498. doi: 10.3760/cma.j.cn112152-20221017-00704.

DOI:10.3760/cma.j.cn112152-20221017-00704

PMID:37355467

Abstract

To investigate the therapeutic effect and mechanism of lenvatinib on regorafenib-resistant hepatocellular carcinoma cells. CCK-8 and clone formation assay were used to observe the inhibitory effect of lenvatinib on the growth of hepatocellular carcinoma cells. Flow cytometry was used to detect the apoptosis of regorafenib-resistant hepatocellular carcinoma cells treated with lenvatinib. The expression levels of related proteins were detected by western blot and immunohistochemical staining. The inhibitory effect of lenvatinib on the tumor formation ability of regorafenib-resistant hepatocellular carcinoma cells in vivo was observed by subcutaneous tumor formation experiment in mice. CCK-8 and clone formation assay showed that lenvatinib could inhibit the proliferation of regorafenib-resistant hepatocellular carcinoma cells. The number of clones of HepG2, SMMC7721 and regorafenib-resistant HepG2, SMMC7721 cells in lenvatinib group (120.67±11.06, 53.00±11.14, 55.00±9.54, 78.67±14.64) were all lower than those in control group (478.00±24.52, 566.00±27.87, 333.67±7.02, 210.00±12.77, all <0.05). Flow cytometry showed that lenvatinib could promote apoptosis of regorafenib-resistant hepatocellular carcinoma cells, the apoptosis rates of HepG2, SMMC7721 and regorafenib-resistant HepG2, SMMC7721 cells in lenvatinib group [(12.30±0.70)%, (9.83±0.38)%, (15.90±1.32)%, (10.60±0.00)%] were all higher than those in control group [(7.50±0.87)%, (5.00±1.21)%, (8.10±1.61)%, (7.05±0.78)%, all <0.05]. The apoptosis-related protein levels suggested that apoptosis was increased in the treatment of lenvatinib. The animal study showed that lenvatinib can inhibit the growth of regorafenib-resistant cells . Immunohistochemistry and western blot results showed that lenvatinib could down-regulate the abnormally activated IGF1R/Mek/Erk signaling pathway in regorafenib-resistant cells. Lenvatinib can reverse regorafenib resistance in hepatocellular carcinoma, possibly by down-regulating IGF1R/Mek/Erk signaling pathway.

摘要

探讨乐伐替尼对瑞戈非尼耐药的肝癌细胞的治疗作用及机制。采用CCK-8法和克隆形成实验观察乐伐替尼对肝癌细胞生长的抑制作用。采用流式细胞术检测乐伐替尼处理的瑞戈非尼耐药肝癌细胞的凋亡情况。通过蛋白质免疫印迹法和免疫组织化学染色检测相关蛋白的表达水平。通过小鼠皮下成瘤实验观察乐伐替尼对瑞戈非尼耐药肝癌细胞体内成瘤能力的抑制作用。CCK-8法和克隆形成实验表明，乐伐替尼可抑制瑞戈非尼耐药肝癌细胞的增殖。乐伐替尼组HepG2、SMMC7721以及瑞戈非尼耐药的HepG2、SMMC7721细胞的克隆数（分别为120.67±11.06、53.00±11.14、55.00±9.54、78.67±14.64）均低于对照组（分别为478.00±24.52、566.00±27.87、333.67±7.02、210.00±12.77，均P<0.05）。流式细胞术显示，乐伐替尼可促进瑞戈非尼耐药肝癌细胞凋亡，乐伐替尼组HepG2、SMMC7721以及瑞戈非尼耐药的HepG2、SMMC7721细胞的凋亡率[分别为(12.30±0.70)%、(9.83±0.38)%、(15.90±1.32)%、(10.60±0.00)%]均高于对照组[分别为(7.50±0.87)%、(5.00±1.21)%、(8.10±1.61)%、(7.05±0.78)%，均P<0.05]。凋亡相关蛋白水平提示乐伐替尼治疗可增加细胞凋亡。动物研究表明，乐伐替尼可抑制瑞戈非尼耐药细胞的生长。免疫组织化学和蛋白质免疫印迹结果显示，乐伐替尼可下调瑞戈非尼耐药细胞中异常激活的IGF1R/Mek/Erk信号通路。乐伐替尼可能通过下调IGF1R/Mek/Erk信号通路逆转肝癌细胞对瑞戈非尼的耐药。