Abstract
The t(4;14)(p16.3;q32) chromosomal translocation occurs in approximately 20% of multiple myelomas (MM) and leads to the apparent deregulation of two genes located on 4p16.3: the fibroblast growth factor receptor 3 (FGFR3) and the putative transcription factor WHSC1/MMSET. Interestingly, FGFR3 mutations known to be associated with autosomal dominant human skeletal disorders have also been found in some MM cell lines with t(4;14) but their pathogenetic role in MM is still controversial. Since cell lines may represent useful models for investigating the effects of deregulated FGFR3 mutants in MM, we analysed the expression, activation, signaling pathways and oncogenic potential of three mutants identified so far: the Y373C and K650E in the KMS-11 and OPM-2 cell lines respectively, and the novel G384D mutation here identified in the KMS-18 cell line. All of the cell lines present a heterozygous FGFR3 gene mutation and transcribe the mutated allele; unlike KMS-11 and OPM-2 (which express the IIIc isoform), the KMS-18 cell line expresses prevalently the isoform IIIb. We demonstrated that, under serum-starved conditions, KMS-11 and OPM-2 cells express appreciable levels of phosphorylated FGFR3 mutants indicating a constitutive activation of the Y373C and K650E receptors; the addition of the aFGF ligand further increased the level of receptor phosphorylation. Conversely, the FGFR3 mutant in KMS-18 does not seem to be constitutively activated since it was phosphorylated only in the presence of the ligand. In all three MM cell lines, ligand-stimulated FGFR3 mutants activated the MAP kinase signaling pathway but did not apparently involve either the STAT1 or STAT3 cascades. However, when transfected in 293T cells, G384D, like Y373C and K650E, was capable of activating MAPK, STAT1 and STAT3 under serum-starved condition. Finally, a focus formation assay of NIH3T3 cells transfected with FGFR3-expressing plasmid vectors showed that Y373C and K650E (albeit at different levels) but not G384D or the wild-type receptor, can induce transformed foci. Overall, our results support the idea that FGFR3 mutations are graded in terms of their activation capability, thus suggesting that they may play a critical role in the tumor progression of MM patients with t(4;14).
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Basilico C, Moscatelli D . 1992 Adv. Cancer Res. 59: 115–165
Cappellen D, De Oliveira C, Ricol D, de Medina SGD, Bourdin J, Sastre-Garau X, Chopin D, Thiery JP, Radvanyl F . 1999 Nature Genet. 23: 18–19
Catlett-Falcone R, Landowski TH, Oshiro MM, Turkson J, Levitzki A, Savino R, Ciliberto G, Moscinski L, Fernandez-Luna JL, Nunez G, Dalton WS, Jove R . 1999 Immunity 16: 2647–2656
Chen C, Okayama H . 1987 Mol. Cell. Biol. 7: 2745–2752
Chesi M, Nardini E, Brents LA, Schrock E, Ried T, Kuel WM, Bergsagel PL . 1997 Nature Genet. 16: 260–264
Chesi M, Nardini E, Lim RSC, Smith KD, Kuel WM, Bergsagel PL . 1998 Blood 92: 3025–3034
d'Avis PY, Robertson SC, Meyer AN, Bardwell WM, Webster MK, Donoghue DJ . 1998 Cell Growth Diff. 9: 71–78
Deng C, Wynshaw-Boris A, Zhou F, Kuo A, Leder P . 1996 Cell 84: 911–921
Finelli P, Fabris S, Zagano S, Baldini L, Intini D, Nobili L, Lombardi L, Maiolo AT, Neri A . 1999 Blood 94: 724–732
Fracchiolla NS, Luminari S, Baldini L, Lombardi L, Maiolo AT, Neri A . 1998 Blood 15: 2987–2989 letter
Francomano CA . 1996 Am. J. Hum. Genet. 59: A25
Greco A, Mariani C, Miranda C, Lupas A, Pagliardini S, Pomati M, Pierotti MA . 1995 Mol. Cell. Biol. 15: 6118–6127
Hart KC, Robertson SC, Kanemitsu MY, Meyer AN, Tynan JA, Donoghue DJ . 2000 Oncogene 19: 3309–3320
Johnson DE, Williams LT . 1993 Adv. Cancer Res. 60: 12–41
Johnson MR, Valentine C, Basilico C, Mansukhani A . 1998 Oncogene 16: 2647–2656
Johnston CL, Cox HC, Gomm JJ, Coombes RC . 1995 J. Biol. Chem. 270: 30643–30650
Kanai M, Goke M, Tsunekawa S, Podolsky DK . 1997 J. Biol. Chem. 272: 6621–6628
Kannan K, Givol D . 2000 Life 49: 197–205
Keegan K, Johnson DE, Williams LT, Hayman M . 1991 Proc. Nat. Acad. Sci. 88: 1095–1099
Legeai-Mallet L, Benoist-Lasselin C, Delezoide AL, Munnich A, Bonaventure J . 1998 J. Biol. Chem. 273: 13007–13014
Li Y, Mangasarian K, Mansukhani A, Basilico C . 1997 Oncogene 14: 1397–1406
Li ZH, Plowright EE, Hawley TS, Bergsagel PL, Chesi M, Hawley RG, Stewart AK . 1998 Blood 92: (suppl.1) A1582
Lofts FJ, Hurst HC, Sternberg MJE, Gullick WJ . 1993 Oncogene 8: 2813–2820
Lombardi L, Newcomb EW, Dalla-Favera R . 1987 Cell 49: 161–170
Malgeri U, Baldini L, Perfetti V, Fabris S, Colli Vignarelli M, Colombo G, Lotti V, Compasso S, Bogni S, Lombardi L, Maiolo AT, Neri A . 2000 Cancer Res. 60: 4058–4061
Muenke M, Schell U . 1997 TIG 11: 308–313
Murgue B, Tsunekawa S, Rosenberg I, deBeaumont M, Podolsky DK . 1994 Cancer Res. 54: 5206–5211
Namba M, Ohtsuki T, Mori M, Togawa A, Wada H, Sugihara T, Yawata Y, Kimoto T . 1989 In Vitro Cell. Devel. Biol. 25: 723–729
Naski M, Wang Q, Xu J, Ornitz DM . 1996 Nature Genet. 13: 233–237
Otsuki T, Nakazawa N, Taniwaki M, Yamada O, Sakaguchi H, Wada H, Yawata Y, Ueki A . 1998 Int. J. Oncol. 12: 545–552
Plowright EE, Li Z, Bergsagel PL, Chesi M, Barber DL, Branch DR, Hawley RG, Stewart AK . 2000 Blood 95: 992–998
Raffioni S, Zhu YZ, Bradshaw RA, Thompson LM . 1998 J. Biol. Chem. 273: 35250–35259
Richelda R, Ronchetti D, Baldini L, Cro L, Viggiano L, Marzella R, Rocchi M, Otsuki T, Lombardi L, Maiolo AT, Neri A . 1997 Blood 90: 4062–4070
Ronchetti D, Bogni S, Finelli P, Lombardi L, Otsuki T, Maiolo AT, Neri A . 2001 Leukemia in press
Rousseau F, Ghouzzi VE, Delezoide AL, Mallet LL, Le Merrer M, Munnich A, Bonaventure J . 1996 Hum. Mol. Genet. 5: 509–512
Sahni M, Ambrosetti DC, Mansukhani A, Gertner R, Levy D, Basilico C . 1999 Genes Dev. 13: 1361–1366
Sambrook J, Fritch EF, Maniatis T . 1989 Molecular Cloning: A Laboratory Manual 2nd Ed Cold Spring Harbour Laboratory Press, Cold Spring Harbour, NY
Scotet E, Houssaint E . 1995 Biochem. Biophys. Acta 1264: 238–242
Stec I, Wright TJ, van Ommen G-JB, de Boer PAJ, van Haeringen A, Moorman AFM, Altherr MR, den Dunnen JT . 1998 Hum. Mol. Genet. 7: 1071–1082
Sternberg MJ, Gullick WJ . 1990 Protein Eng. 3: 245–248
Tavormina PL, Shiang R, Thompson LM, Zhu YZ, Wilkin D, Lachman RS, Wilcox WR, Rimoin DL, Cohn DH, Wasmuth JJ . 1995 Nature Genet. 9: 321–328
Thompson LM, Plummer S, Schalling M, Altherr MR, Gusella JF, Housman DE, Wasmuth JJ . 1991 Genomics 11: 1133–1142
Webster MK, d'Avis PY, Robertson SC, Donoghue DJ . 1996 Mol. Cell. Biol. 16: 4081–4087
Webster MK, Donoghue DJ . 1997a Trends in Genetics 13: 178–182
Webster MK, Donoghue DJ . 1997b Mol. Cell. Biol. 17: 5739–5747
Acknowledgements
Supported by grants from the Associazione Italiana Ricerca sul Cancro (AIRC) (to A Neri), the Ministero della Sanità to Ospedale Maggiore IRCCS, Milan, and from MURST 1999 no 9906038391-010.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ronchetti, D., Greco, A., Compasso, S. et al. Deregulated FGFR3 mutants in multiple myeloma cell lines with t(4;14): comparative analysis of Y373C, K650E and the novel G384D mutations. Oncogene 20, 3553–3562 (2001). https://doi.org/10.1038/sj.onc.1204465
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1204465
Keywords
This article is cited by
-
FLT3-ITD transduces autonomous growth signals during its biosynthetic trafficking in acute myelogenous leukemia cells
Scientific Reports (2021)
-
Coexisting FGFR3 p.K650T mutation in two FGFR3-TACC3 fusion glioma cases
Acta Neuropathologica Communications (2019)
-
Oncogenic signaling by Kit tyrosine kinase occurs selectively on the Golgi apparatus in gastrointestinal stromal tumors
Oncogene (2017)
-
Pan-Raf co-operates with PI3K-dependent signalling and critically contributes to myeloma cell survival independently of mutated RAS
Leukemia (2017)
-
APOBEC family mutational signatures are associated with poor prognosis translocations in multiple myeloma
Nature Communications (2015)