Influence of Stat3 gene repression by RNA interference on metastasis and proliferation of EC-1 cells
AIM: To construct the siRNA expression vector targeting human Stat3 gene and to detect its silencing effects in esophageal carcinoma EC-1 cell lines. METHODS: siRNA expression vectors pSUPER-EGFP-Stat3 were constructed and were transfected into EC-1 cells. The expression of Stat3 mRNA and protein were detected by RT-PCR and Western blot. The invasion ability of EC-1 cells was determined by Boyden-chamber model in vitro and MTT assay was applied to determine the proliferation of EC-1 cells. RESULTS: The siRNA expression vectors pSUPER-EGFP-Stat3 were confirmed by enzyme digestion and DNA sequencing analysis. The results of RT-PCR and Western blot indicated that siRNA knocked down Stat3 mRNA and protein expression. Compared with the EC-1 cells in control groups,Boyden-chamber model showed that the invasion ability of EC-1 cells decreased significantly in interferential group (P < 0.05 ). The level of proliferation also declined in interferential group (P < 0.05). CONCLUSION: siRNA expression vectors inhibit the expression of Stat3 gene effectively in esophageal carcinoma EC-1 cells and suppress the invasion and proliferation ability of EC-1 cells,which lays the foundation for gene therapy of esophageal carcinoma.
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