ABSTRACT
Objective To investigate the role of ECRG2 gene in migration and invasion of human fibrosarcoma HT1080 cells. Methods Tet-on system was used to set up ECRG2-inducible cell clones.Knockdown of ECRG2 gene was done by stably expressing short hairpin RNA (shRNA) specific for ECRG2 mRNA in a highly invasive fibrosarcoma cell line HT1080. The expressions of ECRG2 proteins were detected by Western blotting assay. Wound-healing scrape assay and transwell invasion assay were performed to assess the effect of ECRG2 gene on the invasive properties of HT1080 in vitro. Results The most effective ECRG2siRNA interference and inducible was selected by Western blotting assay. Knockdown of ECRG2 gene significantly promoted the migration and invasion of HT1080 cells while expression of ECRG2 impaired the migration and invasion of HT1080 cells in vitro. Conclusion ECRG2 gene is correlated with the invasive and metastatic potential of fibrosarcoma carcinoma, and expression of ECRG2 induces the inhibition of migration and invasion of HT1080 cell line.
ABSTRACT
Objective To investigate the role of ECRG2,a novel tumor suppressor gene,in spindle assembly checkpoint. Methods Using siRNA approach to deplete the expression of ECRG2, using immunofluorescence to test the distribution of ECRG2,using Western blotting to examine the expression cell cycle proteins.Results ECRG2 localized to centrosomes during interphase and kinetochores during mitosis.Further analysis revealed that ECRG2 participates in the spindle assembly checkpoint.Depletion of ECRG2 abolished the spindle assembly checkpoint.Conclusion Our results indicated that ECRG2 is important for ensuring spindle assembly checkpoint,accurate chromosome segregation,and its depletion may contribute to chmmosome instability and aneuploidy in human cancers.
