Gene Silencing of ß-catenin by RNAi Inhibits Proliferation of Human Esophageal Cancer Cells by Inducing G0/G1 Cell Cycle Arrest


Objectives: The aim of the present study was to explore mechanisms underlying the effects of down-regulatingß-catenin expression on esophageal carcinoma (EC) cells.
Methods: Cell cycle distribution and apoptosis weredetermined using flow cytometry and annexin V apoptosis assay, respectively. Transmission electron microscopy(TEM) was used to examine changes in ultrastructure, while expression of cyclin D1 protein and mRNA wasdetected by western blot and real-time PCR. Proliferating cell nuclear antigen (PCNA) and extracellularsignal-regulated kinase (ERK) 1⁄2 were evaluated by Western blot analysis. PCNA labeling index (LI) wasdetermined by immunocytochemistry.
Results: Compared with pGen-3-con transfected and Eca-109 cells,the percentage of G0/G1-phase pGen-3-CTNNB1 transfected cells was obviously increased (P<0.05), with nosignificant difference among the three groups with regard to apoptosis (P>0.05). pGen-3-CTNNB1 transfectedcells exhibited obvious decrease in cyclin D1 mRNA and protein expression (P<0.05) and the ultrastructure ofEca-109 cells underwent a significant change after being transfected with pGen-3-CTNNB1, suggesting thatdown-regulating β-catenin expression can promote the differentiation and maturation. The expression of PCNAand the ERKI/2 phosphorylation state were also down-regulated in pGen-3-CTNNB1 transfected cells (P<0.05).At the same time, the PCNA labeling index was decreased accordingly (P<0.05).
Conclusion: Inhibition of ECEca-109 cellproliferation by down-regulating ß-catenin expression could improve cell ultrastructure by mediatingblockade in G0/G1 through inhibiting cyclin D1, PCNA and the MAPK pathway (p-ERK1/2).