IKKα/IKKβ/NFκB/SURVIVIN expression regulated by E2F1 in esophageal cancer cell line ECA109

Background: To investigate the regulation of SURVIVIN by E2F1 through the classical NFκB activation pathway and to elucidate the effects of E2F1 on the biological functions of esophageal cancer cells. Materials and methods: An E2F1-overexpression plasmid, pGV142-E2F1, was constructed and transfected into the esophageal cancer cell line ECA109 by electroporation. Transfection efficiency was determined by observing the intensity of green fluorescent protein (GFP) under an inverted fluorescence microscope. Cell cycle and apoptosis of transfected cells were measured by flow cytometry. Total RNA and protein were extracted from transfected cells to determine changes in transcript and protein levels, respectively, of E2F1, IKKα, IKKβ, NFκB, and SURVIVIN. Results: GFP expression in ECA109 cells transfected with the E2F1 overexpression plasmid showed a needle-like shape. The proportion of pGV142-E2F1-transfected ECA109 cells in S-phase was elevated compared to the number of S-phase cells in the empty plasmid and negative control groups (P < 0.05). The proportion of early apoptotic cells following pGV142-E2F1 transfection was significantly reduced compared to that with the empty plasmid and negative control groups (P < 0.05). Expression of E2F1 mRNA and protein were markedly elevated 48 h post-transfection (P < 0.05). Compared to the negative control group, expression levels of NFκB and SURVIVIN mRNA and proteins were significantly increased with overexpression of E2F1. The expression level of IKKα was markedly reduced, and IKKβ mRNA and protein levels were markedly increased on E2F1 overexpression compared to the negative control (P < 0.05). Conclusion: E2F1 and SURVIVIN share a regulatory relationship in ECA109 cells. E2F1 overexpression has a cis-effect on the regulation of IKKβ, NFκB and SURVIVIN, but has a trans-effect on IKKα regulation. Regulation of the expression of SURVIVIN by E2F1 in ECA109 cells may be conducted through the activation of IKKβ, which subsequently activates the classical NFκB activation pathway, leading to the transcriptional activation of SURVIVIN.

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