Chen Tao, Wang Quan, Li Yunxiao, Huang Hefei, Hu Wei
College of Medical Sciences, China Three Gorges University, Yichang, Hubei 443002, P.R. China; Third-Grade Pharmacological Laboratory on Traditional Chinese Medicine Approved by State Administration of Traditional Chinese Medicine, Yichang, Hubei 443002, P.R. China.
Exp Ther Med. 2016 Jun;11(6):2413-2419. doi: 10.3892/etm.2016.3247. Epub 2016 Apr 11.
The aim of the present study was to observe the effects of the Chinese herbal formula QHF (Q, Qingrejiedu; H, Huoxuehuayu; and F, Fuzhengguben) on the migration and invasion of hepatocellular carcinoma (HCC) HepG2 cells and to elucidate the potential molecular mechanisms involved. HepG2 cells were treated with various concentrations of QHF, and scratch and Transwell migration assays were used to qualitatively analyze differences in the migration and invasion activity of these cells. Extracellular signal-regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK) inhibitors were subsequently introduced in order to study the association between QHF and the invasion of HepG2 cells. The protein expression levels of the mitogen-activated protein kinase (MAPK) signaling pathway in HepG2 cells in the presence and absence of QHF were additionally determined using western blot analysis. The results showed that QHF significantly inhibited the proliferation of the HepG2 cells in a concentration-dependent manner, in addition to inhibiting cell movement, which reduced the ability of the cells to invade and migrate. Western blot analysis indicated that the effects of QHF on HCC HepG2 cells after 24 h were to significantly decrease the expression of phosphorylated- (p-)ERK and to increase the expression of p-p38 and p-JNK; however, the total quantity of ERK, p38 and JNK protein remained unchanged. The administration of an inhibitor of ERK altered p38 and JNK expression and promoted the anti-invasion effects of QHF, whereas p38 and JNK inhibitors only partially reversed this effect. The results of the present study indicate, therefore, that QHF is able to inhibit the migratory and invasive activity of HepG2 cells. A possible underlying mechanism involves the activation of the p38 and JNK MAPK signaling pathway and the attenuation of the ERK signaling pathway.
本研究旨在观察中药复方QHF(Q,清热解毒;H,活血化瘀;F,扶正固本)对肝癌HepG2细胞迁移和侵袭的影响,并阐明其潜在的分子机制。用不同浓度的QHF处理HepG2细胞,采用划痕实验和Transwell迁移实验定性分析这些细胞迁移和侵袭活性的差异。随后引入细胞外信号调节激酶(ERK)、p38和c-Jun氨基末端激酶(JNK)抑制剂,以研究QHF与HepG2细胞侵袭之间的关系。此外,通过蛋白质印迹分析测定有无QHF时HepG2细胞中丝裂原活化蛋白激酶(MAPK)信号通路的蛋白质表达水平。结果显示,QHF除抑制细胞运动外,还以浓度依赖的方式显著抑制HepG2细胞的增殖,降低细胞的侵袭和迁移能力。蛋白质印迹分析表明,QHF作用24小时后对肝癌HepG2细胞的影响是显著降低磷酸化(p-)ERK的表达,增加p-p38和p-JNK的表达;然而,ERK、p38和JNK蛋白的总量保持不变。给予ERK抑制剂可改变p38和JNK的表达,并增强QHF的抗侵袭作用,而p38和JNK抑制剂只能部分逆转这种作用。因此,本研究结果表明,QHF能够抑制HepG2细胞的迁移和侵袭活性。一种可能的潜在机制是激活p38和JNK MAPK信号通路并减弱ERK信号通路。