Kim Jaeryeon, Kim Moon-Moo
Department of Chemistry, Dong-Eui University, Busan614-714, Korea.
Department of Applied Chemistry, Dong-Eui University, Busan614-714, Korea.
Curr Mol Pharmacol. 2021 Oct 25;14(4):620-626. doi: 10.2174/1874467213666201005122230.
Anti-cancer effect of lapachol contained in Tabebuia avellandae has been poorly understood until now.
The aim of this study was to investigate the inhibitory effect of lapachol on MMPs related to cell invasion. Its action mechanism was elucidated by analyzing the activity and the expression of MMPs and the proteins involved in the signaling pathway of cell invasion.
The cytotoxicity of lapachol was evaluated by MTT assay in HT1080 cells. The effects of lapachol on the expression and the activation of MMPs were analyzed by western blot, immunofluorescence staining, and gelatin zymography assays. Their gene expression was analyzed by RT-PCR, and metastasis was evaluated by cell invasion assay.
Lapachol below 2 μM showed no cytotoxicity. It was observed that lapachol above 0.5 μM inhibited the activation of MMP-2 and MMP-9 stimulated by PMA. In particular, the protein and gene expression levels of MMP-2 stimulated by PMA were remarkably decreased in the presence of lapachol at 1 μM compared with the PMA treatment group. In addition, lapachol increased the expression level of TIMP-1 compared with the PMA treatment group. Moreover, lapachol decreased the expression level of p-p38 among MAPKs compared with the PMA treatment group. It was also found that the expression level of p65, a part of NF-kB, in nuclei was reduced in the presence of lapachol above 0.5 μM compared with the PMA treatment group. In addition, lapachol inhibited the invasion of human fibrosarcoma cells stimulated with VEGF.
Above results suggest that lapachol could play an important role in the modulation of MMPs related to cell invasion via the increase in TIMP-1 expression as well as the inactivation of p38 through NF-kB transcription factor.
直到现在，人们对杨叶肖槿中所含拉帕醇的抗癌作用了解甚少。
本研究旨在探讨拉帕醇对与细胞侵袭相关的基质金属蛋白酶（MMPs）的抑制作用。通过分析MMPs的活性和表达以及参与细胞侵袭信号通路的蛋白质，阐明其作用机制。
采用MTT法在HT1080细胞中评估拉帕醇的细胞毒性。通过蛋白质印迹法、免疫荧光染色和明胶酶谱分析，分析拉帕醇对MMPs表达和激活的影响。通过逆转录聚合酶链反应（RT-PCR）分析其基因表达，并通过细胞侵袭试验评估转移情况。
2μM以下的拉帕醇未显示出细胞毒性。观察到0.5μM以上的拉帕醇抑制了佛波酯（PMA）刺激的MMP-2和MMP-9的激活。特别是，与PMA处理组相比，在1μM拉帕醇存在下，PMA刺激的MMP-2的蛋白质和基因表达水平显著降低。此外，与PMA处理组相比，拉帕醇增加了金属蛋白酶组织抑制因子-1（TIMP-1）的表达水平。此外，与PMA处理组相比，拉帕醇降低了丝裂原活化蛋白激酶（MAPKs）中磷酸化p38（p-p38）的表达水平。还发现，与PMA处理组相比，在0.5μM以上拉帕醇存在下，细胞核中核因子κB（NF-κB）一部分的p65表达水平降低。此外，拉帕醇抑制了血管内皮生长因子（VEGF）刺激的人纤维肉瘤细胞的侵袭。
上述结果表明，拉帕醇可能通过增加TIMP-1表达以及通过NF-κB转录因子使p38失活，在调节与细胞侵袭相关的MMPs中发挥重要作用。
