The previous studies have showed that neutrophil gelatinase-associated lipocalin (NGAL) was overexpressed in the progress of human esophageal epithelial cells and novel 12-O-tetradecanoyl phorbol-13- acetate (TPA) response elements (TRE) might be located on the -152～-60 position of the 5′ flanking region of the promoter. However, the nucleic proteins bound to the elements have not been identified. In present study, the TRE binding-proteins from EC109 cells following TPA induction were purified by oligonucleotide trapping DNA affinity chromatograph, further separated by SDS-PAGE and stained with silver staining. And then the protein bands were subsequently identified by MALDI-TOF-MS and the resulting spectra were searched to NCBI protein data bank through Mascot. C19, KIAA1949, TDRD1, RXRβ, FAM54A, KLF15, KLF10 and YY-1 eight proteins were identified according to their function, localization and molecular mass. Finally, RT-PCR was performed to determine the TPA responsiveness of these nucleic proteins. Results showed that C19, KIAA1949, TDRD1, RXRβ and KLF15 proteins have the obvious TPA responsiveness in the transcription level. The results suggest these proteins might respond the stimulation of TPA and regulate the expression of NGAL in EC109 cells.