Functional study of calreticulin in oral cancer cells

• Main Authors: Lee-Hsin Wang, 王李馨
• Other Authors: Yi-Fu Chen
• Format: Others
• Language: en_US
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/94752089497759738779

碩士 === 高雄醫學大學 === 生物科技學系碩士班 === 100 === Oral cancer is one of the most common and highly aggressive types of cancer around the world. Among all the oral malignancies, oral squamous cell carcinoma (OSCC) accounts for over 90% of oral malignant tumor. In addition, the five-year survival rate of patients diagnosed with oral cancer is poor in advanced clinical stages due mainly to locoregional spread and distant metastasis. Accordingly, to clarify the mechanisms involved in cancer progression and further identify the biomarkers for OSCC is very important for the early detection and developments of drug and therapeutic methods. We previously analyzed the protein expression profiles between OSCC tumor tissue and normal oral tissue using 2-dimentional electrophoresis and one of the proteins with differential expression was identified as Calreticulin (CRT). CRT is a calcium (Ca2+)-binding chaperone in endoplasmic reticulum (ER) lumen. CRT is a multi-functional protein and has been implicated in the progression of many types of cancer, the function of CRT in OSCC cells remains unclear. Thus, the aim of this study is to investigate the functions of CRT expression in OSCC. We first verified the up-regulated expression of CRT in OSCC tissues and cell lines using western blot and immunohistochemical analyses. To clarify whether CRT is involved in the regulation of cell proliferation and migration, we performed functional assays using transient CRT knockdown cells by siRNA and also CRT RNAi stable line by shRNA. In conclusion, we showed that CRT is overexpressed in OSCC cells, which could be used as a biomarker for OSCC. In addition, the knockdown of CRT expression inhibits cell proliferation, migration and colony formation in OSCC cells. Moreover, we preliminarily explored the downstream target genes and miRNAs using microarray and miRNA array. In the future, we will further identify the CRT-mediated mechanisms involved in the modulation of cell migration.