Ribozyme-mediated targeting of IkappaBgamma inhibits melanoma invasion and metastasis.
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.
Torabian
Sima Z
SZ
Department of Dermatology, University of California San Francisco, San Francisco, CA 94115, USA.
de Semir
David
D
Nosrati
Mehdi
M
Bagheri
Sepideh
S
Dar
Altaf A
AA
Fong
Sylvia
S
Liu
Yong
Y
Federman
Scot
S
Simko
Jeff
J
Haqq
Chris
C
Debs
Robert J
RJ
Kashani-Sabet
Mohammed
M
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
D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D016337:RNA, Catalytic; D012334:RNA, Neoplasm; C073701:Nfkb1 protein, mouse