Ma Yongfang, Liu Xueke, Tang Xiaolong
Medical School, Anhui University of Science and Technology Huainan 232001, Anhui, China.
Department of Clinical Laboratory Medicine, The Fourth Affiliated Hospital, Zhejiang University School of Medicine Yiwu 322000, Zhejiang, China.
Am J Transl Res. 2023 Feb 15;15(2):896-913. eCollection 2023.
The purpose of this study was to clarify the molecular regulatory mechanism of c-Met up-regulated expression and elucidate the molecular mechanisms by which c-Met overexpression and activation drive progression and sorafenib resistance in hepatocellular carcinoma (HCC).
The resistance index was calculated. Bioinformatic techniques were applied to predict the transcription factors that bind and their binding sites on the c-Met promoter. Chromatin immunoprecipitation assays were implemented to verify the prediction results. To determine the regulatory mechanisms and effects of c-Met on sorafenib resistance in HCC, c-Met expression and activation were down-regulated by siRNA and inhibitor in in vivo and vitro experiments, while a parental cell line (Huh-7) was transfected with the adenovirus that upregulated c-Met expression.
c-Met expression was increased in HCC sorafenib-resistant cells. Functional findings suggested that c-Met overexpression and activation drive HCC tumor progression and sorafenib resistance by promoting cell proliferation, migration, and stopping apoptosis. Molecular mechanism findings demonstrated that the MEK/ERK signaling pathway activated the expression and activity of ETS-1 mediated by p-ERK, which led to its binding to the c-Met gene promoter and upregulation of c-Met transcriptional expression. The activation of the HGF/c-Met pathway drives sorafenib resistance in HCC cells by activating the Ras/Raf/ERK and PI3K/Akt signaling pathways, which regulate biologic processes, including cell proliferation, migration and anti-apoptosis.
c-Met overexpression and activation is an essential mechanism of sorafenib resistance in HCC. Combination therapy of sorafenib plus c-Met inhibitor overcame the resistance of sorafenib-targeted therapy for HCC.
本研究旨在阐明c-Met上调表达的分子调控机制，并阐明c-Met过表达和激活驱动肝细胞癌（HCC）进展和索拉非尼耐药的分子机制。
计算耐药指数。应用生物信息学技术预测与c-Met启动子结合的转录因子及其结合位点。实施染色质免疫沉淀试验以验证预测结果。为了确定c-Met对HCC中索拉非尼耐药的调控机制和作用，在体内和体外实验中通过小干扰RNA（siRNA）和抑制剂下调c-Met的表达和激活，同时用上调c-Met表达的腺病毒转染亲本细胞系（Huh-7）。
c-Met在HCC索拉非尼耐药细胞中的表达增加。功能研究结果表明，c-Met过表达和激活通过促进细胞增殖、迁移和抑制凋亡来驱动HCC肿瘤进展和索拉非尼耐药。分子机制研究结果表明，MEK/ERK信号通路激活由p-ERK介导的ETS-1的表达和活性，导致其与c-Met基因启动子结合并上调c-Met转录表达。HGF/c-Met通路的激活通过激活Ras/Raf/ERK和PI3K/Akt信号通路来驱动HCC细胞中的索拉非尼耐药，这些信号通路调节包括细胞增殖、迁移和抗凋亡在内的生物学过程。
c-Met过表达和激活是HCC中索拉非尼耐药的重要机制。索拉非尼联合c-Met抑制剂治疗克服了HCC索拉非尼靶向治疗的耐药性。
