title: Ribozyme-mediated targeting of IkappaBgamma inhibits melanoma invasion and metastasis.
abstract: IkappaBgamma is one member of a family of proteins that can inhibit the nuclear localization of nuclear factor-kappaB. However, the other specific functions of IkappaBgamma are still poorly understood, and its effects on tumor metastasis have not yet been characterized. We examined the consequences of targeting IkappaBgamma in melanoma cells using a hammerhead ribozyme. We developed stable transformant B16-F10 melanoma cell lines that express a ribozyme that targets mouse IkappaBgamma (IkappaBgamma-144-Rz). Tail-vein injection of B16-F10 cells that stably express IkappaBgamma-144-Rz into mice resulted in a significant reduction of the metastatic potential of these cells. IkappaBgamma-144-Rz-expressing B16 cells were shown to have increased transcriptional activity of nuclear factor-kappaB. We then showed that IkappaBgamma-144-Rz-expressing cells demonstrated both reduced invasion and increased apoptosis, suggesting the existence of pathways through which IkappaBgamma promotes melanoma metastasis. Using gene expression profiling, we identified a differentially expressed gene set that is regulated by the stable suppression of IkappaBgamma that may participate in mediating its anti-metastatic effects; we also confirmed the altered expression levels of several of these genes by quantitative real time polymerase chain reaction. Plasmid-mediated expression of IkappaBgamma-144-Rz produced a significant inhibition of the metastatic progression of B16-F10 cells to the lung and resulted in significant anti-invasive and pro-apoptotic effects on murine Lewis lung carcinoma cells. Our results suggest a novel role for IkappaBgamma in promoting the metastatic progression of melanoma.
authors.affiliation: Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA.
authors.forename: Sima Z
authors.initials: SZ
authors.lastname: Torabian
authors.forename: David
authors.initials: D
authors.lastname: de Semir
authors.forename: Mehdi
authors.initials: M
authors.lastname: Nosrati
authors.forename: Sepideh
authors.initials: S
authors.lastname: Bagheri
authors.forename: Altaf A
authors.initials: AA
authors.lastname: Dar
authors.forename: Sylvia
authors.initials: S
authors.lastname: Fong
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Scot
authors.initials: S
authors.lastname: Federman
authors.forename: Jeff
authors.initials: J
authors.lastname: Simko
authors.forename: Chris
authors.initials: C
authors.lastname: Haqq
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Debs
authors.forename: Mohammed
authors.initials: M
authors.lastname: Kashani-Sabet
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D005434:Flow Cytometry; D008546:Melanoma, Experimental / Q000235:genetics* / Q000473:pathology*; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics*; D052002:NF-kappa B p50 Subunit / Q000235:genetics*; D009361:Neoplasm Invasiveness / Q000473:pathology / Q000517:prevention & control*; D009362:Neoplasm Metastasis / Q000473:pathology / Q000517:prevention & control*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D016337:RNA, Catalytic / Q000235:genetics*; D012334:RNA, Neoplasm / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D016337:RNA, Catalytic; D012334:RNA, Neoplasm; C073701:Nfkb1 protein, mouse