Loss of Ubiquitin Activating Enzyme E1 Increases the Sensitivity to 2-Deoxyglucose in Malignant Cancer Cells

• Main Authors: Yen-ChiChen, 陳彥圻
• Other Authors: Wen-Tsan Chang
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/53880180786087346700

碩士 === 國立成功大學 === 生物化學暨分子生物學研究所 === 103 === Ubiquitin is a protein that ubiquitously exists in tissues of eukaryotic organisms. The conjugation of proteins with ubiquitin can regulate many kinds of cellular functions. In many cases, abnormal ubiquitination level in cells causes pathologic reactions, especially the development of cancer. Therefore, the members of ubiquitin pathway could be a great target as novel anticancer drugs. First, I choose the dominant type of E1 enzyme UBE1 as the target of this study. In this research, I establish the stable knockdown UBE1 cancer cell lines by RNAi (RNA interference), including HeLa, H1299, MDA-MB-231, and PC3. The results show that the loss of UBE1 expression decreases cell proliferation rate and cell migration ability. Western blot analysis indicates that many cell cycle proteins like cyclin B1, cdc 2, and cyclin D1 are downregulated in UBE1 knockdown cell. Flow cytometry assay demonstrates that lack of UBE1 causes the change of cell cycle distribution. Furthermore, loss of UBE1 expression causes accumulation of NF-κB inhibitor IκB and inactivation of NF-κB. In pharmacology experiment, I choose three anticancer drugs to analyze the effect on loss of UBE1 expression. In contrast to the mock cells, the sensitivity of these drugs are increased in UBE1 knockdown cells, especially 2-deoxyglucose. The knockdown cells which are treated with low dose 2-deoxyglucose (4 mM) are almost killed. I use the flow cytometry assay to observe cell into apoptosis. Taken together, this study reveals UBE1 can be a potential target for cancer therapy. Key words: Ubiquitin Activating Enzyme E1; 2-Deoxyglucose; Cancer Therapy; Apoptosis