Apigenin inhibits HGF-promoted invasive growth and metastasis involving blocking PI3K/Akt pathway and β4 integrin function in MDA-MB-231 breast cancer cells
Apigenin inhibits HGF-promoted invasive growth and metastasis involving blocking PI3K/Akt pathway and β4 integrin function in MDA-MB-231 breast cancer cells
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DOI：
10.1016/j.taap.2007.09.013
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发表时间：
2008-01-15
影响因子：
3.8
通讯作者：
Tseng, Tsui-Hwa
中科院分区：
文献类型：
--
作者：
Lee, Wei-Jiunn;Chen, Wen-Kang;Tseng, Tsui-Hwa
肝细胞生长因子（HGF）及其受体，Met，已知控制侵入性生长程序，最近已被证明在乳腺癌患者的生存中起着至关重要的作用。膳食来源的黄酮类化合物已被报道具有抗侵袭特性;然而，关于抑制HGF/Met介导的肿瘤侵袭和转移的药理学和分子机制的知识知之甚少。在我们的初步研究中，我们使用HGF作为侵袭诱导剂，研究黄酮类化合物包括芹菜素、柚皮素、染料木素和山萘酚对HGF依赖的MDA-MB-231人乳腺癌细胞侵袭性生长的影响。Boyden小室实验结果表明，芹菜素具有最强的抗迁移和抗侵袭能力。此外，芹菜素抑制HGF诱导的细胞运动和散射，抑制HGF促进的细胞迁移和侵袭，并呈剂量依赖性。免疫印迹分析显示，芹菜素能阻断HGF诱导的Akt磷酸化，但不能阻断Met、ERK和JNK磷酸化。除MDA-MB-231细胞外，芹菜素还对HGF诱导的肝癌SK-Hep 1细胞和肺癌A549细胞Akt磷酸化具有抑制作用。通过间接免疫荧光显微镜分析，芹菜素通过PI 3 K依赖的方式抑制HGF诱导的β 4整合素在富含肌动蛋白的粘附位点和板状伪足的聚集。芹菜素处理抑制HGF刺激的整合素β 4功能，包括MDA-MB-231细胞中的细胞-基质粘附和细胞-内皮细胞粘附。Akt-siRNA转染分析证实芹菜素通过阻断PI 3 K/ Akt通路抑制HGF促进的细胞侵袭性生长。最后，我们通过裸鼠肺移植瘤和鸡胚肺转移瘤的实验研究了芹菜素对HGF促进肿瘤转移的影响。通过小鼠肺组织学和大体检查以及宿主组织中人alu的实时PCR分析，表明芹菜素、渥曼青霉素以及抗β 4抗体都抑制HGF促进的转移。这些数据支持芹菜素通过阻断PI 3 K/Akt通路和整合素β 4功能来抑制HGF促进的侵袭性生长和转移。(c)2007年爱思唯尔公司All rights reserved.
Hepatocyte growth factor (HGF) and its receptor, Met, known to control invasive growth program have recently been shown to play crucial roles in the survival of breast cancer patients. The diet-derived flavonoids have been reported to possess anti-invasion properties;, however, knowledge on the pharmacological and molecular mechanisms in suppressin HGF/Met-mediated tumor invasion and metastasis is poorly understood. In our preliminary study, we use HGF as an invasive inducer to investigate the effect of flavonoids including apigenin, naringenin, genistein and kaempferol on HGF-dependent invasive growth of MDA-MB-231 human breast cancer cells. Results show that apigenin presents the most potent anti-migration and anti-invasion properties by Boyden chamber assay. Furthermore, apigenin represses the HGF-induced cell motility and scattering and inhibits the HGF-promoted cell migration and invasion in a dose-dependent manner. The effect of apigenin on HGF-induced signaling activation involving invasive growth was evaluated by immumoblotting analysis, it shows that apigenin blocks the HGF-induced Akt phosphorylation but not Met, ERK, and JNK phosphorylation. In addition to MDA-MB-231 cells, apigenin exhibits inhibitory effect on HGF-induced Akt phosphorylation in hepatoma SK-Hep1 cells and lung carcinoma A549 cells. By indirect immunofluorescence microscopy assay, apigenin inhibits the HGF-induced clustering of beta 4 integrin at actin-rich adhesive site and lamellipodia through PI3K-dependent manner. Treatment of apigenin inhibited HGF-stimulated integrin beta 4 function including cell-matrix adhesion and cell-endothelial cells adhesion in MDA-MB-231 cells. By Akt-siRNA transfection analysis, it confirmed that apigenin inhibited HGF-promoted invasive growth involving blocking PI3K/ Akt pathway. Finally, we evaluated the effect of apigenin on HGF-promoted metastasis by lung colonization of tumor cells in nude mice and organ metastasis of tumor cells in chick embryo. By histological and gross examination of mouse lung and real-time PCR analysis of human alu in host tissues, it showed that apigenin, wortmannin, as well as anti-beta 4 antibody all inhibit HGF-promoted metastasis. These data support the inhibitory effect of apigenin on HGF-promoted invasive growth and metastasis involving blocking PI3K/Akt pathway and integrin beta 4 function. (c) 2007 Elsevier Inc. All rights reserved.