Regulation of survival, proliferation, invasion, and metastasis of breast cancer cells possibly through modulation of MMPs and Hedgehog signaling pathways by the extract of Zingiber zerumbet — Zerumbone

• Main Authors: Ying-Yu, 陳盈羽
• Other Authors: 張菡馨
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/14104909159291595554

碩士 === 中山醫學大學 === 營養學研究所 === 99 === Breast cancer has ranked first among female cancers in Taiwan and the world. Base on the experiment model using breast cancer lines MDA-MB-468 and MDA-MB-435S, we investigated how the Zerumbone influences the metastasis of breast cancer cell lines. In this study, cell lines were treated with different concentrations of Zerumbone: for MDA-MB-468 were 0, 10, 20uM; as for MDA-MB-435S were 0, 10, 50uM, and we examined the viability of the cells, matrigel invasion assay and migration assay. High concentration of Zerumbone restrained the relative invasion rate (%) of MDA-MB-468, which has reduced 70.15% (P<0.01); moreover, the relative invasion rate of MDA-MB-435S dropped by 95% (P<0.05). Our results suggest Zerumbone effectively decreased the ability of migration of breast cancer cells. Past researches indicated that inflammatory was linked to carcinogenesis, and the chronic long-term inflammatory contributed to cancer. We used iNOS and COX2 antibodys to detect expression of inflammatory, and the result showed that the expression of inflammatory proteins had decreased in MDA-MB-468 with the treatment of Zerumbone 20uM. Therefore, we speculated Zerumbone could inhibit inflammatory. To investigate how Zerumbone inhibit the activity of matrix metalloproteinases and subsequently reduce metastasis, we applied gelatin zymography assay and found out the activity of MMP-2 in conditional medium of MDA-MB-435S decreased; furthermore, analyzing the cell protein we discovered the activity of the two cell lines in MMP-2 demonstrated different trends: from 8 to 16 hours, the MDA-MB-468 shows a descending phenomenon; as for MDA-MB-435S, after 16 hours, the activity of MMP-2 is higher than 8 hours. The relationship between expression of MMP-2 and metastasis was a positive correlation. The signal transduction pathway between Zerumbone and Hedgehog/GLI were related, and we used the Western Blot to examine how the related protein interacted. The result showed Zerumbone can induce the expression of Hedgehog. After 8 and 16 hours treating with Zerumbone, we found tGLI1 (truncated GLI1) expression increased, and strongly correlate with the results of migration. After 24 hours treatment, the expression of GLI1 decreased, while apoptosis increased, providing a possible cause of cell death. We conclude that Zerumbone is able to inhibit the metastasis of human breast cancer cells through different pathways.