College of Light Industry and Food Engineering, Guangxi University, Nanning, China.
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
J Pharm Pharmacol. 2020 Sep;72(9):1165-1175. doi: 10.1111/jphp.13289. Epub 2020 May 17.
Eurycoma longifolia Jack (Simaroubaceae) is commonly distributed in the Southeast Asia and Indo China, which has been shown to possess antianxiety, antibacterial, anticancer, antifungal, anti-inflammatory, antimalarial and antioxidant biological activities. 14,15β-dihydroxyklaineanone is a diterpene isolated from E. longifolia Jack, which is cytotoxic against human lung cancer and human breast cancer cell lines. However, the effects and underlying mechanisms of 14,15β-dihydroxyklaineanone on hepatocellular carcinoma remain unknown.
Cell viability assay and colony formation assay were used to measure HepG2 cell proliferation. Flow cytometry was used to analyse cell cycle and apoptosis. Wound-healing assay and transwell assay were used to observe cells migration. RNA sequencing and the enrichment of differentially expressed genes (DEGs) in Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were used to find and determine underlying pathways.
We found that 14,15β-dihydroxyklaineanone inhibited the growth and migration of HepG2 cells but did not induce cell apoptosis. 14,15β-dihydroxyklaineanone induced S cell cycle arrest by downregulating the expression levels of cyclin A, p-CDK2, cyclin B1, p21, E2F-1 and PCNA. In addition, RNA sequencing showed that 14,15β-dihydroxyklaineanone regulated MAPK pathway by increasing the expression levels of phosphor-p38. Downregulating of p38 via both p38 inhibitor (SB203580) and p38-siRNA could antagonize the inhibition of cell proliferation and migration and reverse the changes in p-p38, E-cadherin, N-cadherin and PCNA expression induced by 14,15β-dihydroxyklaineanone treatment.
14,15β-dihydroxyklaineanone inhibited cell proliferation and migration through regulating p38 MAPK pathway in HCC cells.
长叶勾儿茶（Simaroubaceae）广泛分布于东南亚和印度支那地区，具有抗焦虑、抗菌、抗癌、抗真菌、抗炎、抗疟和抗氧化等生物活性。14,15β-二羟基克拉尼酮是从长叶勾儿茶中分离得到的二萜，对人肺癌和人乳腺癌细胞系具有细胞毒性。然而，14,15β-二羟基克拉尼酮对肝癌的作用及其潜在机制尚不清楚。
采用细胞活力测定和集落形成实验检测 HepG2 细胞的增殖；流式细胞术分析细胞周期和凋亡；划痕实验和 Transwell 实验观察细胞迁移；RNA 测序和京都基因与基因组百科全书（KEGG）通路中差异表达基因（DEGs）的富集用于寻找和确定潜在的通路。
我们发现 14,15β-二羟基克拉尼酮抑制 HepG2 细胞的生长和迁移，但不诱导细胞凋亡。14,15β-二羟基克拉尼酮通过下调细胞周期蛋白 A、p-CDK2、细胞周期蛋白 B1、p21、E2F-1 和 PCNA 的表达水平诱导 S 期细胞周期阻滞。此外，RNA 测序显示 14,15β-二羟基克拉尼酮通过增加磷酸化 p38 的表达水平来调节 MAPK 通路。通过 p38 抑制剂（SB203580）和 p38-siRNA 下调 p38 可以拮抗 14,15β-二羟基克拉尼酮处理诱导的细胞增殖和迁移抑制以及 p-p38、E-钙黏蛋白、N-钙黏蛋白和 PCNA 表达的变化。
14,15β-二羟基克拉尼酮通过调节 HCC 细胞中的 p38 MAPK 通路抑制细胞增殖和迁移。
