Calreticulin Regulates mRNA Stability via Interaction with AU-Rich Element at 3’-Untranslated Region in PC-3 Prostate Cancer Cells

• Main Authors: Chih-Yu Chen, 陳致宇
• Other Authors: Hsinyu Lee
• Format: Others
• Language: en_US
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/2abk5e

碩士 === 國立臺灣大學 === 生命科學系 === 107 === Messenger RNA (mRNA) stability is a post-transcriptional regulation that determines mRNA fate and influences multiple cellular functions. Previous studies have demonstrated that AU-rich element (ARE) in 3’-untranslated region (3’-UTR) of mRNA is critical for mRNA stability via interaction with ARE-binding proteins (ARE-BPs). Chaperone protein calreticulin (CRT) has been reported to bind the 3’-UTR. The expression level of CRT is positively correlated with the expressions of angiogenic factor vascular endothelial growth factor-A (VEGF-A) and β1 integrin. However, the detailed regulatory mechanism of CRT on VEGF-A and β1 integrin expressions remains elusive. Therefore, this study aimed to investigate whether CRT regulates mRNA stability through interaction with ARE. Our results showed that depletion of CRT significantly destabilized VEGF-A and β1 integrin mRNA in PC-3 human prostate cancer cells, and subsequently decreased their expression in both mRNA and protein levels. Sequence analysis showed that ARE in found in the 3’UTR of both mRNA. The result of Luciferase reporter assay demonstrated that CRT interacts with ARE. RNA-immunoprecipitation analysis and electrophoretic mobility shift assay (EMSA) demonstrated that CRT indirectly interacts with mRNA. In summary, this study showed that CRT regulates mRNA stability through interacting with ARE at the 3’-UTR of mRNA.