Identification of protein disulfide bonds affected by ERO1L knockdown in pancreatic cancer cells

• Main Authors: Yu Shiuan Cheng, 程煜瑄
• Other Authors: C. C. Wu
• Format: Others
• Language: zh-TW
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/skt8gk

碩士 === 長庚大學 === 醫學生物技術暨檢驗學系 === 104 === In Taiwan, pancreatic cancer (PC) is ranked tenth among cancer related deaths and shows increased mortality rate in the last decade. To improve the strategy of PC treatment and detection, we have previously identified potential biomarkers and therapeutic targets of PC using integrated analyses of secretome and transcriptome. Among the ten identified candidates, endoplasmic reticulum oxidoreductin 1 like protein (ERO1L) is selected for further validation based on the findings that ERO1L is involved in the protein folding and secretion via the regulation of disulfide bond formation. In this study, for the first time, we revealed that ERO1L level in PC tissues is elevated compared with that in adjacent non-cancerous tissues using immunohistochemistry. We further found that the transwell migration ability of ERO1L-knockdown PC cells, PANC-1 and BxPC-3, was significantly reduced compared to that of the control cells, but the proliferation of the PC cells was not affected by the ERO1L knockdown. To investigate changes of protein disulfide formation in ERO1L-knockdown PC cells, we have established an irreversible isobaric iodoacetyl Cys-reactive tandem mass tag (iodoTMT)-based mass spectrometry (MS) platform. Using this approach, the levels of disulfide bonds in 9 and 12 proteins/peptides are found to be elevated in ERO1L-knockdown PANC-1 and BxPC-3, respectively; meanwhile, 11 and 10 are found to be decreased in ERO1L-knockdown PANC-1 and BxPC-3, respectively. More importantly, among the proteins with dysregulation of disulfide bond formation in the ERO1L-knockdown PC cells, several have been found to play roles in cell migration and cancer metastasis, suggesting that those proteins could involve in the ERO1L-related regulation of cell migration probably via formation of ERO1L-associated disulfide bonds. This postulate is awaiting to be investigated.