Yang Linsong, Ji Weiwei, Zhong Hui, Wang Luyao, Zhu Xiaolin, Zhu Jie
Biomedicine Laboratory, School of Pharmaceutical Engineering and Life Science, Changzhou University Changzhou Jiangsu 213164 P. R. China
Changzhou's Key Laboratory of Pharmaceutical Manufacture and Quality Control Engineering Changzhou 213164 P. R. China.
RSC Adv. 2019 Oct 3;9(54):31517-31526. doi: 10.1039/c9ra06024c. eCollection 2019 Oct 1.
The aim of this paper is to study the anti-tumor mechanism of volatile oil from Thunb. (sodium new houttuyfonate, SNH). , SNH exhibited a concentration-dependent cytotoxic effect against four human cancer lines (HepG2, A2780, MCF-7, SKOV-3). SNH treatment with different concentrations induced HepG2 cells to exhibit varying degrees of morphological changes in apoptotic features, such as round shape, cell shrinkage and formation of apoptotic body. It was observed that SNH caused the decrease in Bcl-2 mRNA expression and triggered the apoptosis of HepG2 cells. Wound healing assay and RT-PCR results showed that the decrease in the expression level of MMP9 and VEGF was observed in HepG2 cells after treatment with SNH for 48 h, suggesting that the extracellular matrix pathway degradation was involved in the HepG2 cells migration. Moreover, we got an insight into the binding mode of SNH into the MMP9 active site through 3D pharmacophore models. Docking study and molecular dynamics (MD) simulation analysis sheds light on that SNH was completely embedded into the MMP9 active site and formed hydrogen bonds with key catalytic residues of MMP9, including Ala191, His190, Ala189 and Glu227. The prediction of SNH binding interaction energies in the MMP9 was almost in good agreement with the original inhibitor EN140. experiments, both SNH and cyclophosphamide significantly reduced tumor weights and their tumor inhibitory rates were 50.78% and 82.61% respectively. This study demonstrated that SNH was an apoptosis inducer in HepG2 cells. SNH has four possible functions, that it could induce apoptosis by mitochondria pathway in HepG2 cells, inhibit the tumor growth, regulate Bcl-2 family mRNA expression and effectively subdue migration of hepatocellular carcinoma cells by decreasing the expression of MMP9 and VEGF. Therefore, SNH might be a potential candidate drug for the treatment of hepatocellular carcinoma, which could provide a reference for further clinical research.
本文旨在研究蕺菜挥发油（新鱼腥草素钠，SNH）的抗肿瘤机制。SNH对四种人类癌细胞系（HepG2、A2780、MCF-7、SKOV-3）表现出浓度依赖性的细胞毒性作用。不同浓度的SNH处理诱导HepG2细胞呈现出不同程度的凋亡特征形态变化，如圆形、细胞收缩和凋亡小体形成。观察到SNH导致Bcl-2 mRNA表达降低并触发HepG2细胞凋亡。伤口愈合实验和RT-PCR结果表明，SNH处理48小时后，HepG2细胞中MMP9和VEGF的表达水平降低，提示细胞外基质途径降解参与了HepG2细胞迁移。此外，我们通过三维药效团模型深入了解了SNH与MMP9活性位点的结合模式。对接研究和分子动力学（MD）模拟分析表明，SNH完全嵌入MMP9活性位点并与MMP9的关键催化残基形成氢键，包括Ala191、His190、Ala189和Glu227。SNH在MMP9中的结合相互作用能预测与原始抑制剂EN140几乎完全一致。实验中，SNH和环磷酰胺均显著降低肿瘤重量，其肿瘤抑制率分别为50.78%和82.61%。本研究表明SNH是HepG2细胞中的凋亡诱导剂。SNH具有四种可能的功能，即它可以通过线粒体途径诱导HepG2细胞凋亡、抑制肿瘤生长、调节Bcl-2家族mRNA表达并通过降低MMP9和VEGF的表达有效抑制肝癌细胞迁移。因此，SNH可能是治疗肝癌的潜在候选药物，可为进一步的临床研究提供参考。
