Effects of HYS-32 on Microtubule-Associated Proteins in Human Glioblastoma Cells

• Main Authors: Hao-Xiang Huang, 黃浩翔
• Other Authors: Jiahn-Chun Wu
• Format: Others
• Language: en_US
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/9s9ah3

碩士 === 國立陽明大學 === 解剖學及細胞生物學研究所 === 104 === HYS-32 containing a cis-stilbene moiety is a novel derivative of combretastatin A4 (CA-4), which can deter microtubule assembly. The ethenyl bridge replaced by 2(5H)-furanones (five-membered aromatic heterocycles) and one of the phenyl rings substituted for naphthalene ring on HYS-32, respectively, can enhance the anti-tumor activity and make better the unstable active cis-stilbene conformation. Previous reports indicated that HYS-32 can against various cancer cell lines and cause microtubule catastrophe in astrocytes. Glioblastoma multiforme (GBM) is considered to be grade IV astrocytoma and has highly aggressive. Metastasis is the primary factor causes 90% death of cancer patients, migration or invasion of cancer cells entrance into blood vessel or lymphatic vessels, transfer to other part of body or organ. In this study, we further investigated the plus end tracking proteins (+TIPs), CLIP-170 and EB1, involved in HYS-32-induced microtubule catastrophe. Glioblastoma cells treated with HYS-32 were processed for immunoblot analysis, phase-contrast microscope, confocal microscopy, and wound healing assay. Phase-contrast microscope showed that HYS-32 (10 µM) induced a morphological change in glioblastoma, from a spindle into a sphere. Confocal microscope with immunofluorescence indicted that HYS-32 induced accumulation of CLIP-170 and eliminated EB1 on microtubule plus ends. HYS-32 treatment did not change the protein levels of CLIP-170 and EB1. However, the levels of β-tubulin were decreased in U87-MG cells (12-24 h) and remained unchanged in U251-MG cells. Treatment of HYS-32 reduced migration ability in U87-MG cells (6-12 h) and U251-MG cells (12 h). Altogether, our observations suggest that HYS-32 causes accumulation of CLIP-170 on microtubule plus end and microtubule catastrophe and inhibits glioblastoma cells migration. HYS-32 can prevent tumor cells metastasis as a new potential therapy.