Department of Biochemistry and Molecular Biology, PR China; Anhui Province Key Laboratory of Active Biological Macro-molecules, PR China.
Anhui Province Key Laboratory of Active Biological Macro-molecules, PR China; School of Clinical Medicine, Wannan Medical College, Wuhu, Anhui 241002, PR China.
J Pharmacol Sci. 2021 Sep;147(1):18-26. doi: 10.1016/j.jphs.2021.05.005. Epub 2021 May 24.
High-mobility group box 1 (HMGB1), a highly conserved chromosome protein, is considered as a potential therapeutic target and novel biomarker because of its regulation in the proliferation and metastasis of Hepatocellular carcinoma (HCC). Calenduloside E (CE), a natural active product, has been reported to anti-cancer effect. However, the role and underlying molecular mechanism of CE in HCC is still unclear. The purpose of this study is to investigate the effects of CE on the proliferation and migration of HCC, and then explore the possible underlying molecular mechanism. HepG2 cells were treated with CE or transfected with HMGB1 shRNA plasmids, EdU and colony formation assays were used to detect cell proliferation ability. Wound healing and transwell assays were used to determine the role of CE in cell migration. The expression of Cyclins, PCNA, MMPs, HMGB1, N-cadherin, E-cadherin and phosphorylation of p38, ERK and JNK were all detected using Western blotting. Our results showed that CE inhibited HepG2 cells proliferation and migration in a dose dependent manner; reduced the expression levels of Cycins, PCNA, HMGB1, MMPs and N-cadherin; up-regulated E-cadherin expression; enhanced the phosphorylation of p38 and JNK signalling pathways. Blocking the activation of p38 and JNK obviously reversed CE-mediated inhibitory effects on HepG2 cell proliferation and migration; reversed CE-induced down-regulation of Cyclins, PCNA, MMPs, N-cadherin and HMGB1, as well as E-cadherin up-regulation. In conclusion, our study suggested that CE reduces the expression levels of Cyclins, MMPs and epithelial-mesenchymal transformation (EMT) through p38/JNK-HMGB1 signaling axis and then inhibits HepG2 cells proliferation and migration in HepG2 cells. This study provides a new perspective for the anti-tumour molecular mechanism of CE in HCC.
高迁移率族蛋白 B1（HMGB1）是一种高度保守的染色体蛋白，因其在肝癌（HCC）的增殖和转移中的调节作用而被认为是潜在的治疗靶点和新型生物标志物。山茱萸苷 E（CE）是一种天然活性产物，已被报道具有抗癌作用。然而，CE 在 HCC 中的作用及其潜在的分子机制尚不清楚。本研究旨在探讨 CE 对 HCC 细胞增殖和迁移的影响，然后探讨其可能的潜在分子机制。用 CE 或 HMGB1 shRNA 质粒转染 HepG2 细胞，用 EdU 和集落形成实验检测细胞增殖能力，用划痕愈合和 Transwell 实验检测 CE 在细胞迁移中的作用。用 Western blot 检测细胞周期蛋白、PCNA、MMPs、HMGB1、N-钙黏蛋白、E-钙黏蛋白和 p38、ERK 和 JNK 磷酸化的表达。结果表明，CE 呈剂量依赖性抑制 HepG2 细胞的增殖和迁移；降低细胞周期蛋白、PCNA、HMGB1、MMPs 和 N-钙黏蛋白的表达水平；上调 E-钙黏蛋白的表达；增强 p38 和 JNK 信号通路的磷酸化。阻断 p38 和 JNK 的激活明显逆转了 CE 介导的对 HepG2 细胞增殖和迁移的抑制作用；逆转了 CE 诱导的细胞周期蛋白、PCNA、MMPs、N-钙黏蛋白和 HMGB1 下调以及 E-钙黏蛋白上调。综上所述，本研究表明，CE 通过 p38/JNK-HMGB1 信号通路降低细胞周期蛋白、MMPs 和上皮间质转化（EMT）的表达水平，从而抑制 HepG2 细胞的增殖和迁移。本研究为 CE 在 HCC 中的抗肿瘤分子机制提供了新的视角。
