Calreticulin regulates vascular endothelial growth factor-A mRNA stability in gastric cancer cells

Autoři: Po-Chu Lee aff001;  Jui-Chung Chiang aff004;  Chih-Yu Chen aff004;  Yin-Chieh Chien aff004;  Wei-Min Chen aff004;  Chin-Wei Huang aff004;  Wen-Chin Weng aff005;  Chia-I Chen aff002;  Po-Huang Lee aff002;  Chiung-Nien Chen aff002;  Hsinyu Lee aff004
Působiště autorů: Department of Traumatology, National Taiwan University Hospital, Taipei, Taiwan aff001;  Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan aff002;  Graduate Institutes of Clinical Medicine, National Taiwan University, Taipei, Taiwan aff003;  Department of Life Science, National Taiwan University, Taipei, Taiwan aff004;  Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan aff005;  Department of Pediatrics, National Taiwan University, Taipei, Taiwan aff006;  Department of Pediatric Neurology, National Taiwan University Children’s Hospital, Taipei, Taiwan aff007;  Department of Surgery, E-DA Hospital, Kaohsiung, Taiwan aff008;  Department of Electrical Engineering, National Taiwan University, Taipei, Taiwan aff009;  Angiogenesis Research Center, National Taiwan University, Taipei, Taiwan aff010;  Research Center for Developmental Biology, National Taiwan University, Taipei, Taiwan aff011;  Regenerative Medicine and Center for Biotechnology, National Taiwan University, Taipei, Taiwan aff012
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0225107


Calreticulin (CRT) and vascular endothelial growth factor-A (VEGF-A) are crucial for angiogenesis, and mediate multiple malignant behaviors in gastric cancer. In this study, we report that CRT is positively correlated with VEGF-A in gastric cancer patients. Moreover, high expressions of both CRT and VEGF-A are markedly associated with the pathological stage, progression, and poor prognosis in the patients. Therefore, we sought to elucidate the mechanism by which CRT affects VEGF-A in gastric cancer. Firstly, we demonstrate the novel finding that knockdown of CRT reduced VEGF-A mRNA stability in two gastric cancer cell lines, AGS and MKN45. The AU-Rich element (ARE) is believed to play a crucial role in the maintenance of VEGF-A mRNA stability. Luciferase reporter assay shows that knockdown of CRT significantly decreased the activity of renilla luciferase with VEGF-A ARE sequence. Additionally, competition results from RNA-binding/electrophoretic mobility shift assay indicate that CRT forms an RNA-protein complex with the VEGF-A mRNA by binding to the ARE. In addition, the proliferation rate of human umbilical vein endothelial cells (HUVEC) was significantly reduced when treated with conditioned medium from CRT knockdown cells; this was rescued by exogenous VEGF-A recombinant protein. Our results demonstrate that CRT is involved in VEGF-A ARE binding protein complexes to stabilize VEGF-A mRNA, thereby promoting the angiogenesis, and progression of gastric cancer.

Klíčová slova:

Angiogenesis – Gastric cancer – Immunoprecipitation – Luciferase – Messenger RNA – Metastasis – RNA extraction – RNA-binding proteins


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