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Cellular Oncology
Published in: Cellular Oncology 3/2014

01-06-2014 | Original Paper

Identification of GPM6A and GPM6B as potential new human lymphoid leukemia-associated oncogenes

Authors: Cyndia Charfi, Elsy Edouard, Eric Rassart

Published in: Cellular Oncology | Issue 3/2014

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Abstract

Background

Previously, we found that the Graffi murine leukemia virus (MuLV) is able to induce a wide spectrum of hematologic malignancies in vivo. Using high-density oligonucleotide microarrays, we established the gene expression profiles of several of these malignancies, thereby specifically focusing on genes deregulated in the lymphoid sub-types. We observed over-expression of a variety of genes, including Arntl2, Bfsp2, Gfra2, Gpm6a, Gpm6b, Nln, Fbln1, Bmp7, Etv5 and Celsr1 and, in addition, provided evidence that Fmn2 and Parm-1 may act as novel oncogenes. In the present study, we assessed the expression patterns of eight selected human homologs of these genes in primary human B-cell malignancies, and explored the putative oncogenic potential of GPM6A and GPM6B.

Methods

The gene expression levels of the selected human homologs were tested in human B-cell malignancies by semi-quantitative RT-PCR. The protein expression profiles of human GPM6A and GPM6B were analyzed by Western blotting. The localization and the effect of GPM6A and GPM6B on the cytoskeleton were determined using confocal and indirect immunofluorescence microscopy. To confirm the oncogenic potential of GPM6A and GPM6B, classical colony formation assays in soft agar and focus forming assays were used. The effects of these proteins on the cell cycle were assessed by flow cytometry analysis.

Results

Using semi-quantitative RT-PCR, we found that most of the primary B-cell malignancies assessed showed altered expression patterns of the genes tested, including GPM6A and GPM6B. Using confocal microscopy, we found that the GPM6A protein (isoform 3) exhibits a punctate cytoplasmic localization and that the GPM6B protein (isoform 4) exhibits a peri-nuclear and punctate cytoplasmic localization. Interestingly, we found that exogenous over-expression of both proteins in NIH/3T3 cells alters the actin and microtubule networks and induces the formation of long filopodia-like protrusions. Additionally, we found that these over-expressing NIH/3T3 cells exhibit anchorage-independent growth and enhanced proliferation rates. Cellular transformation (i.e., loss of contact inhibition) was, however, only observed after exogenous over-expression of GPM6B.

Conclusions

Our results indicate that several human homologs of the genes found to be deregulated in Graffi MuLV experimental mouse models may serve as candidate biomarkers for human B-cell malignancies. In addition, we found that GPM6A and GPM6B may act as novel oncogenes in the development of these malignancies.
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Literature
1.
2.
V. Voisin, C. Barat, T. Hoang, E. Rassart, Novel insights into the pathogenesis of the Graffi murine leukemia retrovirus. J. Virol. 80, 4026–4037 (2006)PubMedCentralPubMedCrossRef
3.
C. Denicourt, E. Edouard, E. Rassart, Oncogene activation in myeloid leukemias by Graffi murine leukemia virus proviral integration. J. Virol. 73, 4439–4442 (1999)PubMedCentralPubMed
4.
C. Denicourt, C.A. Kozak, E. Rassart, Gris1, a new common integration site in Graffi murine leukemia virus-induced leukemias: overexpression of a truncated cyclin D2 due to alternative splicing. J. Virol. 77, 37–44 (2003)PubMedCentralPubMedCrossRef
5.
C. Denicourt, P. Legault, F.A. McNabb, E. Rassart, Human and mouse cyclin D2 splice variants: transforming activity and subcellular localization. Oncogene 27, 1253–1262 (2008)PubMedCrossRef
6.
C. Charfi, V. Voisin, L.C. Levros Jr., E. Edouard, E. Rassart, Gene profiling of Graffi murine leukemia virus-induced lymphoid leukemias: identification of leukemia markers and Fmn2 as a potential oncogene. Blood 117, 1899–1910 (2011)PubMedCrossRef
7.
C. Charfi, L.C. Levros Jr., E. Edouard, E. Rassart, Characterization and identification of PARM-1 as a new potential oncogene. Mol. Cancer 12, 84 (2013)PubMedCentralPubMedCrossRef
8.
X. Castells, J.J. Acebes, S. Boluda, A. Moreno-Torres, J. Pujol, M. Julia-Sape, A.P. Candiota, J. Arino, A. Barcelo, C. Arus, Development of a predictor for human brain tumors based on gene expression values obtained from two types of microarray technologies. OMICS 14, 157–164 (2010)PubMedCrossRef
9.
P. Urban, M. Bilecova-Rabajdova, Z. Stefekova, A. Ostro, M. Marekova, Overview of potential oncomarkers for detection of early stages of ovarian cancer. Klin. Onkol. 24, 106–111 (2011)PubMed
10.
S. Landais, S. Landry, P. Legault, E. Rassart, Oncogenic potential of the miR-106-363 cluster and its implication in human T-cell leukemia. Cancer Res. 67, 5699–5707 (2007)PubMedCrossRef
11.
M. Ru, C. Shustik, E. Rassart, Graffi murine leukemia virus: molecular cloning and characterization of the myeloid leukemia-inducing agent. J. Virol. 67, 4722–4731 (1993)PubMedCentralPubMed
12.
E. Edouard, C. Charfi, V. Voisin, C. Denicourt, C. Barat, E. Rassart, The Graffi murine leukemia virus: a multipotent retrovirus ideal for the study and characterization of lymphoid and non lymphoid leukemias. Transw. Res. Netw. 8, 23–47 (2012)
13.
V. Voisin, P. Legault, D.P. Ospina, Y. Ben-David, E. Rassart, Gene profiling of the erythro- and megakaryoblastic leukaemias induced by the Graffi murine retrovirus. BMC Med. Genomics 3, 2 (2010)PubMedCentralPubMedCrossRef
14.
F. Sievers, A. Wilm, D. Dineen, T.J. Gibson, K. Karplus, W. Li, R. Lopez, H. McWilliam, M. Remmert, J. Soding, J.D. Thompson, D.G. Higgins, Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol. Syst. Biol. 7, 539 (2011)PubMedCentralPubMedCrossRef
15.
K.C. Chou, H.B. Shen, Signal-CF: a subsite-coupled and window-fusing approach for predicting signal peptides. Biochem. Biophys. Res. Commun. 357, 633–640 (2007)PubMedCrossRef
16.
N.H. Colburn, W.F. Bruegge, J.R. Bates, R.H. Gray, J.D. Rossen, W.H. Kelsey, T. Shimada, Correlation of anchorage-independent growth with tumorigenicity of chemically transformed mouse epidermal cells. Cancer Res. 38, 624–634 (1978)PubMed
17.
V.H. Freedman, S.I. Shin, Cellular tumorigenicity in nude mice: correlation with cell growth in semi-solid medium. Cell 3, 355–359 (1974)PubMedCrossRef
18.
S.I. Shin, V.H. Freedman, R. Risser, R. Pollack, Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc. Natl. Acad. Sci. U. S. A. 72, 4435–4439 (1975)PubMedCentralPubMedCrossRef
19.
20.
M. Ikeda, W. Yu, M. Hirai, T. Ebisawa, S. Honma, K. Yoshimura, K.I. Honma, M. Nomura, cDNA cloning of a novel bHLH-PAS transcription factor superfamily gene, BMAL2: its mRNA expression, subcellular distribution, and chromosomal localization. Biochem. Biophys. Res. Commun. 275, 493–502 (2000)PubMedCrossRef
21.
22.
Y. Yasuniwa, H. Izumi, K.Y. Wang, S. Shimajiri, Y. Sasaguri, K. Kawai, H. Kasai, T. Shimada, K. Miyake, E. Kashiwagi, G. Hirano, A. Kidani, M. Akiyama, B. Han, Y. Wu, I. Ieiri, S. Higuchi, K. Kohno, Circadian disruption accelerates tumor growth and angio/stromagenesis through a Wnt signaling pathway. PLoS ONE 5, e15330 (2010)PubMedCentralPubMedCrossRef
23.
G. Mazzoccoli, V. Pazienza, A. Panza, M.R. Valvano, G. Benegiamo, M. Vinciguerra, A. Andriulli, A. Piepoli, ARNTL2 and SERPINE1: potential biomarkers for tumor aggressiveness in colorectal cancer. J. Cancer Res. Clin. Oncol. 138, 501–511 (2012)PubMedCrossRef
24.
S.D. Georgatos, F. Gounari, S. Remington, The beaded intermediate filaments and their potential functions in eye lens. BioEssays 16, 413–418 (1994)PubMedCrossRef
25.
M.D. Perng, Q. Zhang, R.A. Quinlan, Insights into the beaded filament of the eye lens. Exp. Cell Res. 313, 2180–2188 (2007)PubMedCrossRef
26.
M.A. Aldahmesh, A.O. Khan, J. Mohamed, F.S. Alkuraya, Novel recessive BFSP2 and PITX3 mutations: insights into mutational mechanisms from consanguineous populations. Genet. Med. 13, 978–981 (2011)PubMedCrossRef
27.
G. Wu, L. Zhou, L. Khidr, X.E. Guo, W. Kim, Y.M. Lee, T. Krasieva, P.L. Chen, A novel role of the chromokinesin Kif4A in DNA damage response. Cell Cycle 7, 2013–2020 (2008)PubMedCentralPubMedCrossRef
28.
Y. Kurasawa, W.C. Earnshaw, Y. Mochizuki, N. Dohmae, K. Todokoro, Essential roles of KIF4 and its binding partner PRC1 in organized central spindle midzone formation. EMBO J. 23, 3237–3248 (2004)PubMedCentralPubMedCrossRef
29.
M. Mazumdar, J.H. Lee, K. Sengupta, T. Ried, S. Rane, T. Misteli, Tumor formation via loss of a molecular motor protein. Curr. Biol. 16, 1559–1564 (2006)PubMedCentralPubMedCrossRef
30.
Y.S. Kim, J.D. Hwan, S. Bae, D.H. Bae, W.A. Shick, Identification of differentially expressed genes using an annealing control primer system in stage III serous ovarian carcinoma. BMC Cancer 10, 576 (2010)PubMedCentralPubMedCrossRef
31.
L.L. Yates, C. Schnatwinkel, J.N. Murdoch, D. Bogani, C.J. Formstone, S. Townsend, A. Greenfield, L.A. Niswander, C.H. Dean, The PCP genes Celsr1 and Vangl2 are required for normal lung branching morphogenesis. Hum. Mol. Genet. 19, 2251–2267 (2010)PubMedCentralPubMedCrossRef
32.
M. Katoh, Comparative integromics on non-canonical WNT or planar cell polarity signaling molecules: transcriptional mechanism of PTK7 in colorectal cancer and that of SEMA6A in undifferentiated ES cells. Int. J. Mol. Med. 20, 405–409 (2007)PubMed
33.
C. Korz, A. Pscherer, A. Benner, D. Mertens, C. Schaffner, E. Leupolt, H. Dohner, S. Stilgenbauer, P. Lichter, Evidence for distinct pathomechanisms in B-cell chronic lymphocytic leukemia and mantle cell lymphoma by quantitative expression analysis of cell cycle and apoptosis-associated genes. Blood 99, 4554–4561 (2002)PubMedCrossRef
34.
A.W. Fjorback, H.K. Muller, O. Wiborg, Membrane glycoprotein M6B interacts with the human serotonin transporter. J. Mol. Neurosci. 37, 191–200 (2009)PubMedCrossRef
35.
S. Olinsky, B.T. Loop, A. DeKosky, B. Ripepi, W. Weng, J. Cummins, S.L. Wenger, Y. Yan, C. Lagenaur, V. Narayanan, Chromosomal mapping of the human M6 genes. Genomics 33, 532–536 (1996)PubMedCrossRef
36.
N.L. Nadon, S. Miller, K. Draeger, M. Salvaggio, Myelin proteolipid DM20: evidence for function independent of myelination. Int. J. Dev. Neurosci. 15, 285–293 (1997)PubMedCrossRef
37.
X. Castells, J.M. Garcia-Gomez, A. Navarro, J.J. Acebes, O. Godino, S. Boluda, A. Barcelo, M. Robles, J. Arino, C. Arus, Automated brain tumor biopsy prediction using single-labeling cDNA microarrays-based gene expression profiling. Diagn. Mol. Pathol. 18, 206–218 (2009)PubMedCrossRef
38.
Y. Yan, C. Lagenaur, V. Narayanan, Molecular cloning of M6: identification of a PLP/DM20 gene family. Neuron 11, 423–431 (1993)PubMedCrossRef
39.
H. Werner, L. Dimou, M. Klugmann, S. Pfeiffer, K.A. Nave, Multiple splice isoforms of proteolipid M6B in neurons and oligodendrocytes. Mol. Cell. Neurosci. 18, 593–605 (2001)PubMedCrossRef
40.
J.L. Popot, D. Pham Dinh, A. Dautigny, Major myelin proteolipid: the 4-alpha-helix topology. J. Membr. Biol. 120, 233–246 (1991)PubMedCrossRef
41.
T. Weimbs, W. Stoffel, Proteolipid protein (PLP) of CNS myelin: positions of free, disulfide-bonded, and fatty acid thioester-linked cysteine residues and implications for the membrane topology of PLP. Biochemistry 31, 12289–12296 (1992)PubMedCrossRef
42.
A. Gow, A. Gragerov, A. Gard, D.R. Colman, R.A. Lazzarini, Conservation of topology, but not conformation, of the proteolipid proteins of the myelin sheath. J. Neurosci. 17, 181–189 (1997)PubMed
43.
A.M. Schneider, I.R. Griffiths, C. Readhead, K.A. Nave, Dominant-negative action of the jimpy mutation in mice complemented with an autosomal transgene for myelin proteolipid protein. Proc. Natl. Acad. Sci. U. S. A. 92, 4447–4451 (1995)PubMedCentralPubMedCrossRef
44.
E. Swanton, A. Holland, S. High, P. Woodman, Disease-associated mutations cause premature oligomerization of myelin proteolipid protein in the endoplasmic reticulum. Proc. Natl. Acad. Sci. U. S. A. 102, 4342–4347 (2005)PubMedCentralPubMedCrossRef
45.
S. Yang, W. Wang, C. Lei, Q. Liu, F. Xu, X. Xing, H. Chen, J. Liu, S. Wu, M. Wang, Localization and characterization of rat transmembrane protein 225 specifically expressed in testis. DNA Cell Biol. 30, 9–16 (2011)PubMedCentralPubMedCrossRef
46.
M. Namiki, M. Kitamura, E. Buczko, M.L. Dufau, Rat testis P-450(17)alpha cDNA: the deduced amino acid sequence, expression and secondary structural configuration. Biochem. Biophys. Res. Commun. 157, 705–712 (1988)PubMedCrossRef
47.
J. Haugstetter, T. Blicher, L. Ellgaard, Identification and characterization of a novel thioredoxin-related transmembrane protein of the endoplasmic reticulum. J. Biol. Chem. 280, 8371–8380 (2005)PubMedCrossRef
48.
M. Schweneker, A.S. Bachmann, K. Moelling, JM4 is a four-transmembrane protein binding to the CCR5 receptor. FEBS Lett. 579, 1751–1758 (2005)PubMedCrossRef
49.
M. Wang, G. Luo, F. Li, S. Wu, J. Jiang, C. Huang, Isolation and characterization of a novel four-transmembrane protein PMP22CD specifically expressed in the testis. Int. J. Mol. Sci. 7, 425–437 (2006)CrossRef
50.
H. Ohno, J. Stewart, M.C. Fournier, H. Bosshart, I. Rhee, S. Miyatake, T. Saito, A. Gallusser, T. Kirchhausen, J.S. Bonifacino, Interaction of tyrosine-based sorting signals with clathrin-associated proteins. Science 269, 1872–1875 (1995)PubMedCrossRef
51.
52.
B.D. Trapp, A. Nishiyama, D. Cheng, W. Macklin, Molecular bases for the recognition of tyrosine-based sorting signals. J. Cell Biol. 137, 459–468 (1997)PubMedCentralPubMedCrossRef
53.
M.P. Sinoway, K. Kitagawa, S. Timsit, G.A. Hashim, D.R. Colman, Proteolipid protein interactions in transfectants: implications for myelin assembly. J. Neurosci. Res. 37, 551–562 (1994)PubMedCrossRef
54.
J. Alfonso, M.E. Fernandez, B. Cooper, G. Flugge, A.C. Frasch, The stress-regulated protein M6a is a key modulator for neurite outgrowth and filopodium/spine formation. Proc. Natl. Acad. Sci. U. S. A. 102, 17196–17201 (2005)PubMedCentralPubMedCrossRef
55.
B. Fuchsova, M.E. Fernandez, J. Alfonso, A.C. Frasch, Cysteine residues in the large extracellular loop (EC2) are essential for the function of the stress-regulated glycoprotein M6a. J. Biol. Chem. 284, 32075–32088 (2009)PubMedCentralPubMedCrossRef
56.
M.A. Brocco, M.E. Fernandez, A.C. Frasch, Filopodial protrusions induced by glycoprotein M6a exhibit high motility and aids synapse formation. Eur. J. Neurosci. 31, 195–202 (2010)PubMedCrossRef
57.
K.Y. Huang, G.D. Chen, C.H. Cheng, K.Y. Liao, C.C. Hung, G.D. Chang, P.P. Hwang, S.Y. Lin, M.C. Tsai, K.H. Khoo, M.T. Lee, C.J. Huang, Phosphorylation of the zebrafish M6Ab at serine 263 contributes to filopodium formation in PC12 cells and neurite outgrowth in zebrafish embryos. PLoS ONE 6, e26461 (2011)PubMedCentralPubMedCrossRef
58.
M.E. Fernandez, J. Alfonso, M.A. Brocco, A.C. Frasch, Conserved cellular function and stress-mediated regulation among members of the proteolipid protein family. J. Neurosci. Res. 88, 1298–1308 (2010)PubMed
59.
F. Xue, D.M. Janzen, D.A. Knecht, Contribution of filopodia to cell migration: a mechanical link between protrusion and contraction. Int. J. Cell Biol. 2010, 507821 (2010)PubMedCentralPubMed
60.
A. Arjonen, R. Kaukonen, J. Ivaska, Filopodia and adhesion in cancer cell motility. Cell Adhes. Migr. 5, 421–430 (2011)CrossRef
61.
H. Li, S. Hou, X. Wu, S. Nandagopal, F. Lin, S. Kung, A.J. Marshall, The tandem PH domain-containing protein 2 (TAPP2) regulates chemokine-induced cytoskeletal reorganization and malignant B cell migration. PLoS ONE 8, e57809 (2013)PubMedCentralPubMedCrossRef
62.
C. Scielzo, M.T. Bertilaccio, G. Simonetti, A. Dagklis, E. ten Hacken, C. Fazi, M. Muzio, V. Caiolfa, D. Kitamura, U. Restuccia, A. Bachi, M. Rocchi, M. Ponzoni, P. Ghia, F. Caligaris-Cappio, HS1 has a central role in the trafficking and homing of leukemic B cells. Blood 116, 3537–3546 (2010)PubMedCrossRef
Metadata
Title
Identification of GPM6A and GPM6B as potential new human lymphoid leukemia-associated oncogenes
Authors
Cyndia Charfi
Elsy Edouard
Eric Rassart
Publication date
01-06-2014
Publisher
Springer Netherlands
Published in
Cellular Oncology / Issue 3/2014
Print ISSN: 2211-3428
Electronic ISSN: 2211-3436
DOI
https://doi.org/10.1007/s13402-014-0171-y

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