This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zhu, L. Articles by Yen, Y. Search for Related Content PubMed PubMed Citation Articles by Zhu, L. Articles by Yen, Y.

¹ Departments of Medical Oncology and Therapeutic Research and ² Cytogenetics, City of Hope National Medical Center, Duarte, California and ³ Department of Basic Medical Science, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China

Requests for reprints: Yun Yen, Department of Medical Oncology and Therapeutic Research, City of Hope National Medical Center, 1500 East Duarte Road, Duarte, CA 91010. Phone: 626-359-8111, ext. 62867; Fax: 626-301-8233. E-mail: yyen{at}coh.org

The presence of t(4;14)(p16.3;q32.3) in multiple myeloma cells results in dysregulated expression of the fibroblast growth factor receptor 3 (FGFR3). FGFR3 acts as an oncogene to promote multiple myeloma cell proliferation and antiapoptosis. These encourage the clinical development of FGFR3-specific inhibitors. Three short hairpin RNAs (shRNA) targeting different sites of FGFR3 were selected and subsequently transfected into KMS-11, OPM-2, and NCI-H929 human myeloma cell lines, all of which are characterized by t(4;14) and FGFR3 over expression. The combination of these three shRNAs can effectively inhibit FGFR3 expression in all three cell lines. Sequential immunocytochemistry/fluorescence in situ hybridization was employed to validate that the shRNAs specifically inhibited FGFR3 expression in OPM-2 cells. Decreased expression of B-cell chronic lymphocytic leukemia/lymphoma 2 (BCL2) and myeloid cell leukemia sequence 1 (MCL1) proteins and increased staining of Annexin V–positive cells showed that inhibition of FGFR3 induces apoptosis. After confirming down-regulation of FGFR3 by real-time PCR, HU-133 plus 2.0 array was employed to compare the gene expression profile of shRNA-treated sample with that of the control. Besides the down-regulation of FGFR3, expression of the antiapoptotic genes CFLAR, BCL2, MCL1, and some members of NF-B family decreased, whereas expression of the proapoptotic genes CYC, BID, CASP2, and CASP6 increased. Microarray results also revealed changes in genes previously implicated in multiple myeloma pathogenesis (RAS, RAF, IL-6R, and VEGF), as well as others (TLR4, KLF4, and GADD45A) not previously linked to multiple myeloma. Our observations indicate that shRNAs can specifically and effectively inhibit FGFR3 expression. This targeted approach may be worth testing in multiple myeloma patients with t(4;14) and FGFR3 overexpression in the future.

Key Words: short hairpin RNA • fibroblast growth factor receptor 3 • multiple myeloma • t(4;14) • apoptosis

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Received 12/ 8/04; revised 2/17/05; accepted 3/15/05.

This article has been cited by other articles:

				J. Lauring, A. M. Abukhdeir, H. Konishi, J. P. Garay, J. P. Gustin, Q. Wang, R. J. Arceci, W. Matsui, and B. H. Park The multiple myeloma associated MMSET gene contributes to cellular adhesion, clonogenic growth, and tumorigenicity Blood, January 15, 2008; 111(2): 856 - 864. [Abstract] [Full Text] [PDF]

				V. Bedell, S. J. Forman, K. Gaal, V. Pullarkat, L. M. Weiss, and M. L. Slovak Successful Application of a Direct Detection Slide-Based Sequential Phenotype/Genotype Assay Using Archived Bone Marrow Smears and Paraffin Embedded Tissue Sections J. Mol. Diagn., November 1, 2007; 9(5): 589 - 597. [Abstract] [Full Text] [PDF]

				S. Arnaud-Dabernat, M. Kritzik, A. G. Kayali, Y.-Q. Zhang, G. Liu, C. Ungles, and N. Sarvetnick FGFR3 Is a Negative Regulator of the Expansion of Pancreatic Epithelial Cells Diabetes, January 1, 2007; 56(1): 96 - 106. [Abstract] [Full Text] [PDF]

				E. Masih-Khan, S. Trudel, C. Heise, Z. Li, J. Paterson, V. Nadeem, E. Wei, D. Roodman, J. O. Claudio, P. L. Bergsagel, et al. MIP-1{alpha} (CCL3) is a downstream target of FGFR3 and RAS-MAPK signaling in multiple myeloma Blood, November 15, 2006; 108(10): 3465 - 3471. [Abstract] [Full Text] [PDF]

				J. Ren, D. Raina, W. Chen, G. Li, L. Huang, and D. Kufe MUC1 Oncoprotein Functions in Activation of Fibroblast Growth Factor Receptor Signaling Mol. Cancer Res., November 1, 2006; 4(11): 873 - 883. [Abstract] [Full Text] [PDF]

				S. Trudel, A. K. Stewart, E. Rom, E. Wei, Z. H. Li, S. Kotzer, I. Chumakov, Y. Singer, H. Chang, S.-B. Liang, et al. The inhibitory anti-FGFR3 antibody, PRO-001, is cytotoxic to t(4;14) multiple myeloma cells Blood, May 15, 2006; 107(10): 4039 - 4046. [Abstract] [Full Text] [PDF]