title: Regulation of neutrophil gelatinase-associated lipocalin expression by C/EBPβ in lung carcinoma cells.
abstract: Neutrophil gelatinase-associated lipocalin (NGAL), a member of the lipocalin family, has been found to be overexpressed in a variety of tumors, including lung adenocarcinomas. However, the mechanism by which NGAL expression is regulated in lung carcinoma needs further evaluation. In this study, immunohistochemistry was employed to analyze the expression of NGAL in lung carcinoma tissue samples, including lung squamous carcinomas, adenocarcinomas, adenosquamous carcinomas and bronchial alveolar cell carcinomas. The results showed that NGAL was expressed in 82.61% (19/23) of the samples. RT-PCR and immunofluorescent staining showed that NGAL was localized to the cytoplasm in lung carcinoma cell lines. To explore the transcriptional regulation mechanism of NGAL basal expression in lung carcinoma, a 1515-bp fragment (-1431 to +84) of the NGAL promoter region was cloned and a series of deletion and mutation constructs were generated. These constructs were analyzed using the luciferase reporter assay. The results indicated that the cis-acting elements important for the basal activity of NGAL transcription were likely located between -152 and -141. Further analysis using site-directed mutagenesis and the luciferase reporter assay suggested that the C/EBP binding sites were responsible for the activity of the NGAL promoter. Finally, the binding ability and specificity of the transcription factors were determined by electrophoretic mobility-shift assay (EMSA). The results showed that C/EBPβ was able to bind to the -152 and -141 segments. Taken together, these findings suggest that NGAL is expressed in lung carcinomas and that NGAL expression is mediated by the binding of C/EBPβ to the -152 and -141 segment of the NGAL promoter.
authors.affiliation: Department of Biochemistry and Molecular Biology;
authors.forename: Pi-Xian
authors.initials: PX
authors.lastname: Zhang
authors.forename: Jing-Xia
authors.initials: JX
authors.lastname: Chang
authors.forename: Jian-Jun
authors.initials: JJ
authors.lastname: Xie
authors.forename: Hua-Min
authors.initials: HM
authors.lastname: Yuan
authors.forename: Ze-Peng
authors.initials: ZP
authors.lastname: DU
authors.forename: Fa-Ren
authors.initials: FR
authors.lastname: Zhang
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Lü
authors.forename: Li-Yan
authors.initials: LY
authors.lastname: Xu
authors.forename: En-Min
authors.initials: EM
authors.lastname: Li1