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

English version

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: https://doi.org/10.1371/journal.pone.0225107

Souhrn

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

Zdroje

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

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