To investigate the functions of Numb in oral carcinogenesis using gene knockout technology

• Main Authors: Chun-Yu Fan Chiang, 范姜群譽
• Other Authors: Kuo-Wei Chang
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/z9247f

碩士 === 國立陽明大學 === 口腔生物研究所 === 105 === Oral squamous cell carcinoma (OSCC) is one of the common cancers worldwide. Numb is an highly conserved protein and it displays a complex pattern of functions. Recent studies have shown Numb as an oncogenic suppressor by inhibiting proliferation, migration, stemness properties, and others. We found in this study that Numb mRNA expression was down-regulated in a large fraction of OSCC tumors. We established Numb 1 and Numb 4 expression lentiviral constructs, and the gene edition Numb-knockout (KO) OSCC cell subclones using CRISPR Cas9 system. The analysis showed that the anchorage independent growth of OSCC cells was elevated after deletion of Numb, while such ability was repressed after the rescue of the Numb expression by viral infection. In addition, the level of lactate production and the ratio of ECAR to OCR in the Numb-KO cells was higher than parental cells. Reports showed that monocarboxylate transporter (MCT) family members were responsible for transporting lactate out of cells. This study further found that the deletion of Numb was able to up-regulate MCT1 and MCT4 expression to increase the lactate production. Advance studies are undergoing to clarify the interplay between anchorage independent growth potential and MCT1/4 expression modulated by Numb in OSCC.