Chen Chien-Min, Hsieh Shu-Ching, Lin Chia-Liang, Lin Yu-Syun, Tsai Jen-Pi, Hsieh Yi-Hsien
Division of Neurosurgery, Department of Surgery, Changhua Christian Hospital, Changhua, Taiwan.
School of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Cell Physiol Biochem. 2017;44(4):1460-1470. doi: 10.1159/000485582. Epub 2017 Dec 1.
BACKGROUND/AIMS: α-mangostin has anti-carcinogenic effects against several cancers. We investigated the molecular mechanism of this compound on the metastasis of human renal carcinoma cells.
Cell viability was measured using the MTT assay, and cell cycle distribution using flow cytometry. A Matrigel-based assay was used to measure in vitro cell migration and invasion. MAPK-related proteins and matrix metalloproteinase (MMP)-9 and MMP-2 expression were measured by western blotting, and MMP2/-9 activities were determined by gelatin zymography. RT-qPCR and a luciferase assay were used to examine the transcriptional activity of MMP-9.
α-mangostin inhibited the migration and invasion of RCC cells in a dose-dependent manner, but had no evident cytotoxic effects. Treatment of 786-O cells with α-mangostin inhibited activation of MEK and ERK. Treatment with a specific MEK inhibitor (U0126) enhanced the inhibitory effects of α-mangostin on cell migration and invasion, and the phosphorylation of ERK and MEK. Moreover, α-mangostin inhibited the expression of the MMP-9 mRNA levels as well as the activity of MMP-9 promoter, and these suppressive effects were further enhanced by U0126.
Our results suggest that α-mangostin suppresses cell migration and invasion via MEK/ERK/MMP9 pathway, and might be a promising anti-metastatic agent against human renal cell carcinoma.
背景/目的：α-山竹黄酮对多种癌症具有抗癌作用。我们研究了该化合物对人肾癌细胞转移的分子机制。
使用MTT法测定细胞活力，采用流式细胞术检测细胞周期分布。基于基质胶的实验用于测量体外细胞迁移和侵袭。通过蛋白质印迹法检测MAPK相关蛋白以及基质金属蛋白酶（MMP）-9和MMP-2的表达，通过明胶酶谱法测定MMP2/-9的活性。使用RT-qPCR和荧光素酶测定法检测MMP-9的转录活性。
α-山竹黄酮以剂量依赖性方式抑制肾癌细胞的迁移和侵袭，但无明显细胞毒性作用。用α-山竹黄酮处理786-O细胞可抑制MEK和ERK的激活。用特异性MEK抑制剂（U0126）处理可增强α-山竹黄酮对细胞迁移和侵袭的抑制作用，以及ERK和MEK的磷酸化。此外，α-山竹黄酮抑制MMP-9 mRNA水平的表达以及MMP-9启动子的活性，而U0126可进一步增强这些抑制作用。
我们的结果表明，α-山竹黄酮通过MEK/ERK/MMP9途径抑制细胞迁移和侵袭，可能是一种有前景的抗人肾细胞癌转移药物。
