Investigating the role of SOX2 in gastric carcinoma cell line MKN45

• Main Authors: PEI-PEI JHAN, 詹珮佩
• Other Authors: Chin Li
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/77zg8f

碩士 === 國立中正大學 === 生命科學系分子生物研究所 === 105 === In 2014, gastric cancer is the seventh and tenth most common cancer in male and female, respectively. It is a cancer incurring high mortality. In all, 3768 patients were diagnosed, and the mortality was 2241. Despite of its high mortality, our understanding on the molecular pathways of gastric epithelia cell differentiation and tumorigenesis is still not complete. SOX2, a specific transcription factor during stomach development, maintains gastric epithelial cell phenotype and preserves stem cell pluripotency. Aberrant down-regulation of SOX2 expression has been reported to be involved in gastric cancer pathogenesis. To investigate the function of SOX2 in gastric cancer cell lines, we knock-outed SOX2 by CRISPR/CAS9 system and overexpressed SOX2 by Lentivirus system in MKN45 cells. The effect of complete SOX2 depletion and overexpression to growth and migration of MKN45 was then examined. The result showed that MKN45/koSOX2 or MKN45/FLAG-SOX2 exhibited similar growth rates and colony formation ability as MKN45/CTL. However, MKN45/koSOX2 had faster migration rate in would healing assay but MKN45/FLAG-SOX2 exhibits similar migration to MKN45/CTL. Since MKN45 expressed endogenous SOX2, we will use SOX2-negative gastric cancer line MKN28 for following overexpression experiment. In addition, we profiled global gene expression change through next generation sequencing to identify candidate SOX2-regulated genes. We are currently confirming the results of expression profiling and will investigate the functions of the SOX2-regulated genes in gastric carcinoma.