Abstract:

Objective To discuss the inhibitory effect of chelerythrine （CHE） on the migration， invasion， and epithelial-mesenchymal transition （EMT） of the human ovarian cancer SKOV3 cells，and to clarify the associated mechanism. Methods The SKOV3 cells were cultured in vitro and divided into control group and 2.5， 5.0， 10.0， 20.0， and 40.0 μmol·L^－1 CHE groups.Methylthiazolydiphenyl-tetrazolium（MTT） assay was used to detect the inhibitory rates of proliferation of the cells in various groups. The SKOV3 cells were cultured in vitro and divided into control group， transforming growth factor-β1 （TGF-β1） group， TGF-β1+5 μmol·L^－1 CHE group， and TGF-β1+10 μmol·L^－1 CHE group.Cell scratch assay was used to detect the migration rates of the cells in various groups； Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups； Western blotting method was used to detect the expression levels of E-cadherin， N-cadherin， and Vimentin proteins in the cells in various groups； immunofluorescence staining method was used to detect the fluorescence intensities of E-cadherin and N-cadherin in the cells in various groups. Results The MTT assay results showed that compared with control group， the inhibitory rates of proliferation of the cells in 5.0， 10.0， 20.0， and 40.0 μmol·L^－1 CHE groups were significantly increased （P<0.05 or P<0.01）. The cell scratch assay results showed that compared with control group， the migration rate of the cells in TGF-β1 group was increased （P<0.01）； compared with TGF-β1 group， the migration rates of the cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly decreased （P<0.01）. The Transwell chamber assay results showed that compared with control group， the numbers of migration and invasion cells in TGF-β1 group were significantly increased （P<0.05）； compared with TGF-β1 group， the numbers of migration and invasion cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly decreased （P<0.01）. The Western blotting results showed that compared with control group， the expression level of E-cadherin protein in the cells in TGF-β1 group was significantly decreased （P<0.01）， while the expression levels of N-cadherin and Vimentin proteins were increased （P<0.05 or P<0.01）； compared with TGF-β1 group， the expression levels of E-cadherin protein in the cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly increased （P<0.01）， and the expression levels of N-cadherin and Vimentin proteins were significantly decreased （P<0.01）. The immunofluorescence staining results showed that compared with control group， the fluorescence intensity of E-cadherin in the cells in TGF-β1 group was decreased， and the fluorescence intensity of N-cadherin was increased； compared with TGF-β1 group， the fluorescence intensities of E-cadherin in the cells in TGF-β1+5 μmol·L^－1 CHE group and TGF-β1+10 μmol·L^－1 CHE group were significantly increased， and the fluorescence intensities of N-cadherin were decreased. Conclusion CHE can inhibit the proliferation， migration， invasion， and EMT of the human ovarian cancer SKOV3 cells.

Key words: Chelerythrine, Ovarian neoplasm, Epithelial-mesenchymal transition, Transforming growth factor-β1, Cell migration, Cell invasion