Gene expression profile by inhibiting Raf-1 protein kinase in breast cancer cells

  • Authors:
    • Rajshree R. Mewani
    • Song Tian
    • Bihua Li
    • Malika T. Danner
    • Theresa D. Carr
    • Sung Lee
    • Aquilur Rahman
    • Usha N. Kasid
    • Mira Jung
    • Anatoly Dritschilo
    • Prafulla C. Gokhale
  • View Affiliations

  • Published online on: March 1, 2006     https://doi.org/10.3892/ijmm.17.3.457
  • Pages: 457-463
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Abstract

Raf-1 protein serine-threonine kinase plays an important role in cell growth, proliferation, and cell survival. Previously, we and others have demonstrated that antisense raf oligonucleotide-mediated inhibition of Raf-1 expression leads to tumor growth arrest, radiosensitization and chemosensitization in vivo. Raf-1 inhibition is also associated with apoptotic cell death. In this study, we inhibited Raf-1 using an antisense raf oligonucleotide (AS-raf-ODN) to identify downstream targets of Raf-1 using microarray gene expression analysis. Treatment of MDA-MB-231 breast cancer cells with 250 nM AS-raf-ODN led to significant inhibition of Raf-1 protein (75.2±9.6%) and c-raf-1 mRNA levels (86.2±3.3%) as compared to untreated control cells. The lipofectin control or mismatch oligonucleotide had no effect on Raf-1 expression. To determine the changes in gene expression profiles that were due to inhibition of Raf-1, we simultaneously compared the gene expression patterns in AS-raf-ODN treated cells with untreated control cells and cells treated with lipofectin alone or MM-ODN. A total of 17 genes (4 upregulated and 13 down-regulated) including c-raf-1 were identified that were altered after AS-raf-ODN treatment. Functional clustering analysis revealed genes involved in apoptosis (Bcl-XL), cell adhesion (paxillin, plectin, Rho GDIα, CCL5), metabolism (GM2A, SLC16A3, PYGB), signal transduction (protein kinase C nu), and transcriptional regulation (HMGA1), and membrane-associated genes (GNAS, SLC16A3). Real-time PCR, Northern analysis and Western analysis confirmed the microarray findings. Our study provides insight into Raf-1 related signaling pathways and a model system to identify potential target genes.

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March 2006
Volume 17 Issue 3

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Mewani RR, Tian S, Li B, Danner MT, Carr TD, Lee S, Rahman A, Kasid UN, Jung M, Dritschilo A, Dritschilo A, et al: Gene expression profile by inhibiting Raf-1 protein kinase in breast cancer cells. Int J Mol Med 17: 457-463, 2006
APA
Mewani, R.R., Tian, S., Li, B., Danner, M.T., Carr, T.D., Lee, S. ... Gokhale, P.C. (2006). Gene expression profile by inhibiting Raf-1 protein kinase in breast cancer cells. International Journal of Molecular Medicine, 17, 457-463. https://doi.org/10.3892/ijmm.17.3.457
MLA
Mewani, R. R., Tian, S., Li, B., Danner, M. T., Carr, T. D., Lee, S., Rahman, A., Kasid, U. N., Jung, M., Dritschilo, A., Gokhale, P. C."Gene expression profile by inhibiting Raf-1 protein kinase in breast cancer cells". International Journal of Molecular Medicine 17.3 (2006): 457-463.
Chicago
Mewani, R. R., Tian, S., Li, B., Danner, M. T., Carr, T. D., Lee, S., Rahman, A., Kasid, U. N., Jung, M., Dritschilo, A., Gokhale, P. C."Gene expression profile by inhibiting Raf-1 protein kinase in breast cancer cells". International Journal of Molecular Medicine 17, no. 3 (2006): 457-463. https://doi.org/10.3892/ijmm.17.3.457