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¹ Department of Pathology, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA
² Department of Pathology, University of Michigan Medical School and Comprehensive Cancer Center, University of Michigan, Ann Arbor, Michigan 48109-0054, USA
³ Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA
⁴ Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania, USA

(Requests for offprints should be addressed to Y E Nikiforov who is now at Department of Pathology, University of Pittsburgh, A713 Scaife Hall, 3550 Terrace Street, Pittsburgh, Pennsylvania 15261, USA; Email: nikiforovye{at}upmc.edu)

R Ciampi is now at Department of Endocrinology and Metabolism, University of Pisa, Pisa, Italy

	Abstract

Top Abstract Introduction Materials and methods Results Discussion References

 
Chromosomal rearrangements of the RET proto-oncogene (RET/PTC) are the common feature of papillary thyroid carcinoma (PTC). In this study, we report the identification, cloning, and functional characterization of a novel type of RET/PTC rearrangement that results from the fusion of the 3'-portion of RET coding for the tyrosine kinase (TK) domain of the receptor to the 5'-portion of the Homo sapiens hook homolog 3 (HOOK3) gene. The novel fusion was identified in a case of PTC that revealed a gene expression signature characteristic of RET/PTC on DNA microarray analysis, but was negative for the most common types of RET rearrangement. A fusion product between exon 11 of HOOK3 and exon 12 of RET gene was identified by 5'RACE, and the presence of chimeric HOOK3-RET protein of 88 kDa was detected by western blot analysis with an anti-RET antibody. The protein is predicted to contain a portion of the coiled-coil domains of HOOK3 and the intact TK domain of RET. Expression of the HOOK3-RET cDNA in NIH3T3 cells resulted in the formation of transformed foci and in tumor formation after injection into nude mice, confirming the oncogenic nature of HOOK3-RET.

	Introduction

Top Abstract Introduction Materials and methods Results Discussion References

 
Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer and accounts for ~80% of all thyroid malignancies (Hundahl et al. 2000). Genetic alterations along the RET/RAS/BRAF/ MAPK signaling pathway have been demonstrated to be crucial in the pathogenesis of papillary carcinomas, as they are found in more than 70% of all tumors and rarely overlap in the same tumor, suggesting that activation of a single effector of this pathway is sufficient for cell transformation (Kimura et al. 2003, Soares et al. 2003, Frattini et al. 2004). The most common genetic alteration of this pathway is the point mutation V600E of the BRAF gene, which is found in ~40% of papillary carcinomas (reviewed in Ciampi & Nikiforov 2005, Xing 2005). Recently, BRAF has also been shown to be activated in papillary carcinomas by chromosomal rearrangement resulting in the AKAP9-BRAF fusion (Ciampi et al. 2005). It is a rare event in sporadic papillary carcinomas and more common in radiation-induced tumors. Approximately, 10% of papillary carcinomas harbor point mutations in one of the RAS genes (N-RAS, H-RAS, K-RAS), with most of these tumors being the follicular variants of papillary carcinoma (De Micco 2003, Zhu et al. 2003). Chromosomal rearrangements involving the RET proto-oncogene is another common alteration in PTC (Fusco et al. 1987, Grieco et al. 1990).

The RET proto-oncogene resides on chromosome 10q11.2 and codes for a membrane receptor tyrosine kinase (TK; Takahashi et al. 1985, Takahashi 1988). It contains three functional domains: an extracellular ligand-binding domain with four cadherin-like repeats and one cysteine-rich region, a hydrophobic trans-membrane domain, and an intracellular domain containing the TK domain. The ligands of the RET receptor belongs to the group of the glial cell line-derived neutrophic factor family and include neurturin (NRTN), persephin (PSPN), and artemin (ARTN; Airaksinen et al. 1999). Binding of the ligand causes receptor dimerization, autophosphorylation of critical tyrosine residues within the intracellular domain, and activation of the signaling cascade.

In the thyroid gland, RET is expressed at high level in parafollicular C-cells, where point mutations of RET are responsible for the development of medullary thyroid carcinoma. In follicular thyroid cells, RET can be activated by chromosomal rearrangement resulting in the fusion of the 3'-portion of the RET gene to the 5'-portion of several unrelated genes, which is known as RET/PTC rearrangement (Fusco et al. 1987, Grieco et al. 1990). Three types of RET/PTC were originally identified and remain by far the most common in papillary carcinomas. Of them, RET/PTC1 is formed by fusion with the H4 (also known as D10S170, CCDC6) gene (Grieco et al. 1990) and RET/PTC3 by fusion with the NCOA4 (RFG, ELE1) gene (Bongarzone et al. 1994, Santoro et al. 1994). RET/PTC1 and RET/PTC3 are intrachromosomal paracentric inversions since both genes participating in the rearrangement are located on chromosome 10q (Pierotti et al. 1992, Minoletti et al. 1994). In contrast, RET/PTC2 is a translocation between chromosomes 10 and 17, resulting in the RET fusion with the regulatory subunit RI of the cAMP-dependent protein kinase A (Bongarzone et al. 1993). More recently, several novel types of RET/PTC have been described in single cases of PTC, all of which are interchromosomal translocations (Klugbauer et al. 1998, 2000, Klugbauer & Rabes 1999, Nakata et al. 1999, Corvi et al. 2000, Salassidis et al. 2000, Saenko et al. 2003).

In a previous study of PTC using DNA microarray analysis, we determined a distinct expression profile for the RET/PTC rearrangement and established a highly accurate mutational classifier based on the expression of several genes (Giordano et al. 2005). We identified a case of papillary carcinoma in which the classifier predicted the presence of RET/PTC rearrangement, despite the negative testing of the tumor RNA for RET/PTC1 and RET/PTC3 rearrangements by RT-PCR. By fluorescence in situ hybridization (FISH), we demonstrated that the tumor cells indeed harbor a RET rearrangement, but one that did not correspond to the most common RET/PTC1 or RET/PTC3 types. In this study, we performed further analysis of this case which resulted in the identification, cloning, and functional characterization of a novel type of RET/PTC rearrangement.

	Materials and methods

Top Abstract Introduction Materials and methods Results Discussion References

 
Tumor samples and cell lines

Snap frozen tissue from the index case was obtained from the University of Michigan Health System through the Tissue Procurement Service with Institutional Review Board (IRB) approval. The index case was a 14-year-old female with a 7 x 4 x 4 cm papillary carcinoma of the right lobe. The tumor morphology was classical papillary type and there were four positive central compartment lymph nodes. There was no history of prior radiation exposure. Frozen tissue samples from 129 PTC were obtained from the Department of Pathology, University of Cincinnati following a protocol approved by the University of Cincinnati IRB and through the Cooperative Human Tissue Network.

Fluorescence in situ hybridization (FISH)

FISH was performed as previously described (Ciampi et al. 2005). Briefly, a mix of the two bacterial artificial chromosome (BAC) clones RP11-168L22 (GenBank accession no. AC068707) and RP13-368N15 (GenBank accession no. AL591169) were used to design a probe spanning the RET gene. The clone RP11-122A17 (GenBank accession no. AC110275) was used as a probe for the HOOK3 gene. The clones were purchased from BAC/PAC Resources, Children’s Hospital, Oakland. The extracted DNA was labeled with Spectrum-Green-dUTP and SpectrumRed-dUTP using a nick translation kit (Vysis Inc., Downers Grove, IL, USA). Touch-preparations from the tumor were prepared from snap frozen tissue and fixed in 3:1 methanol:acetic acid, pretreated with 10 mg collagenase H (Sigma), and co-denatured with 10 ng each labeled probe. Hybridization was carried at 37 °C overnight. Microscopy was performed with a Leica Microsystem TCS 4D confocal fluorescence microscope with digital image capture (Leica, Wetzlar, Germany).

5'RACE

Total RNA from the index case was extracted using Trizol Reagent (Invitrogen) and subjected to 5'RACE using the 5'RACE System for Amplification of cDNA Ends (Invitrogen). cDNA synthesis was performed using a previously published primer corresponding to exon 13 of the RET gene, 5'-CTGCTTCAGGACGTTGAA-3' (Santoro et al. 1994). Subsequent PCRs utilized primer 5'-GCAGGTCTCCGCAGCTCACTC-3' for the first reaction and 5'-CTTTCAGCATCTTCACGG-3' (Santoro et al. 1994) for the second round of amplification. The RACE products were excised from the gel, purified and sequenced using an automated ABI 377 Sequencer (Perkin–Elmer, Waltham, MA, USA).

Cloning and sequencing analysis

The open reading frame (ORF) of the chimeric cDNA was amplified using primers spanning the start codon of HOOK3 (5'-CACCATGTTCAGCGTAGAG-3') and the stop codon of RET (5'-ACTATCAAACGTGTCCATTAATTT-3'). The PCR product was electrophoresed on agarose gel, purified, and cloned into the pcDNA3.1D/V5-His-TOPO expression vector using the pcDNA3.1 Directional TOPO Expression kit (Invitrogen). Several clones were fully sequenced and the clone I-9 was chosen for further experiments.

RT-PCR experiments

Total RNA was extracted using Trizol Reagent (Invitrogen), reverse transcribed, and amplified by PCR using primers flanking the HOOK3-RET usion point: 5'-AGGCGGCAGGTTAAACTCTT-3' located in the exon 11 of HOOK3 and 5'-TCCAAATTCGCCTTCTCCTA-3' in the exon 12 of RET, with an expected product of 195 bp. Amplification was carried out for 35 cycles (94 °C for 40 s, 55 °C for 1 min, and 72 °C for 1 min 30 s). In order to study the expression of the wild-type HOOK3, semi-quantitative RT-PCR was performed for 30 cycles using primers 5'-TCCAAATTCGCCTTCTCCTA-3' and 5'-TGTTCTCAGCCTGTCCTTTTC-3', with the expected amplification product of 246 bp. The results were normalized using amplification of the housekeeping gene phosphoglycerate kinase (PGK), as described elsewhere (Argani et al. 1998).

Western blot analysis

Western blotting was performed as previously described (Kunzli et al. 2002). Twenty microgram of proteins were loaded on a 7.5% polyacrylamide gel and subjected to SDS–PAGE. After the electro-transfer, membranes were incubated with anti-RET mouse monoclonal antibody (clone RET-X-D210) generated against the C-terminus of the RET protein in our laboratory and previously validated (Knauf et al. 2003). The dilution of the antibody was 1:250. HRP-goat-anti-mouse secondary antibody (Zymed, San Francisco, CA, USA) was used for the detection. Protein extracts from liver, medullary thyroid carcinoma, and TPC1 cell line harboring RET/PTC1 rearrangement (Ishizaka et al. 1990) were used as controls.

Transformation assays

NIH 3T3 cells were plated in six-well plates at a density of 3.4 x 10⁵ and transfected 24 h later using Lipofectamine (Invitrogen) with 16 µg pcDNA3.1D HOOK3-RET plasmid. Cells transfected with the empty vector were used as a negative control. After 1 day, cells were split into three 100 mm³ dishes and cultured in DMEM with 10% fetal bovine serum (FBS). For the focus assay, cells were cultured for 21 days, stained with 0.5% crystal violet, and foci counted. For stable cell lines, cells were mass selected for 21 days in the media supplemented with 350 µg/ml G-418 Sulfate (Invitrogen). A total of 5 x 10⁶ pooled cells were suspended in 0.4 ml culture media and then injected subcutaneously into the flank of homozygous Nu/Nu female mice (Charles River Laboratories, Wilmington, MA, USA). Cells expressing HOOK3-RET were injected into the left flank and cells containing empty vector were injected into the right flank of each of the six mice. Tumors were removed 20–26 days later, measured, weighed, and fixed in 10% formalin for histologic evaluation.

	Results

Top Abstract Introduction Materials and methods Results Discussion References

 
We studied a case of papillary carcinoma that had revealed an expression profile consistent with RET/PTC rearrangement in our previous study (Giordano et al. 2005). The presence of RET rearrangement in the majority of tumor cells was confirmed with FISH using a probe spanning the RET gene. A split of one of the two RET signals was noted in the majority of tumor cells (Fig. 1A).

View larger version (46K): [in this window] [in a new window]   Figure 1 Fluorescence in situ hybridization (FISH) on interphase tumor cell nuclei from the index case. (A) Hybridization with the RET probe only shows split of one of the signals (arrows) indicating the presence of the RET gene rearrangement. (B) Two-color FISH with the RET probe (green) and the HOOK3 probe (red) confirming the HOOK-RET fusion (arrow). Full color version available at http://erc.endocrinology-journals.org.

View larger version (26K): [in this window] [in a new window]   Figure 2 (A) Sequence of the 5' RACE product showing the fusion between HOOK3 and RET. (B) Genomic organization of the HOOK3 and RET genes and the HOOK3-RET fusion. The boxes indicate exons and the lines indicate introns. Black arrows indicate the location of breakpoint in the two genes. The chimeric cDNA is shown at the bottom of the panel with indication of its size (bp) and the number of corresponding amino acids.

View larger version (28K): [in this window] [in a new window]   Figure 3 Expression of the wild-type HOOK3 gene in thyroid and other tissues. Electrophoresis of RT-PCR products showing HOOK3 expression in the liver (L), skeletal muscle (SM), kidney (K), and three samples from normal thyroid (NT). PGK amplification was used as a control for RNA quantity. M, 123 bp DNA ladder; NC, negative control.

View larger version (72K): [in this window] [in a new window]   Figure 4 Western blot analysis with the RET specific antibody of protein extracts from liver (A), medullary thyroid carcinoma (B), TPC1 cell line (C), and in the index case of papillary carcinoma (D). The index case shows a band corresponding to predicted molecular weight of 88.2 kDa of the HOOK3-RET protein (*). TPC1 cell line harboring RET/PTC1 rearrangement shows an expected band corresponding to 60 kDa (**), while medullary thyroid carcinoma shows two bands at 170 and 180 kDa corresponding to the wild-type RET (***). Liver cell extract as expected does not reveal the RET protein. An additional band (arrow) is likely to be due to non-specific antibody binding.

The ability of HOOK3-RET protein to induce cell transformation was studied in an NIH3T3 cells focus assay. Cells transfected with the HOOK3-RET ORF cDNA formed transfected foci at high efficiency when compared with cells transformed with empty vector (Fig. 5A). In addition, NIH3T3 cells stably transfected with HOOK3-RET formed tumors in nude mice as early as 10 days after injection. Tumors formed in all mice injected and grew to 0.78–1.22 g (20–26 mm in size) 3 weeks after injection. In contrast, no tumors were detected after injection of cells transfected with empty vector (Fig. 5B). These results demonstrate that HOOK3-RET functions as an oncogene and is able to transform NIH3T3 cells both in vitro and in vivo.

View larger version (7K): [in this window] [in a new window]   Figure 5 (A) Formation of transformed foci in NIH3T3 cells transiently transfected with pcDNA3.1D/HOOK3-RET and the empty vector. The results represent the average number of foci per plate in three separate plates. Similar results were obtained in two separate experiments. (B) s.c. injection of NIH3T3 cells stably transfected with HOOK3-RET or empty vector in the flank of nude mice. Results are shown as average weight of tumors removed from six mice.

	Discussion

Top Abstract Introduction Materials and methods Results Discussion References

 
In this study, we report the identification of a novel RET/PTC rearrangement that results from fusion of the RET and HOOK3 genes. Since the identification of the first RET partner in a chromosomal rearrangement reported in 1990 (Grieco et al. 1990), 12 different types of RET/PTC have been identified resulting from the fusion of RET to different partner genes (Table 1; Bongarzone et al. 1993, 1994, Santoro et al. 1994, Klugbauer et al. 1998, 2000, Klugbauer & Rabes 1999, Nakata et al. 1999, Corvi et al. 2000, Salassidis et al. 2000, Saenko et al. 2003). RET/PTC1 and RET/PTC3 result from intrachromosomal inversions, and constitute the two most common RET/PTC rearrangement types found in sporadic and radiation-associated papillary carcinomas. RET/PTC2 and the more recently identified rare types of RET/PTC are all interchromosomal translocations. Most of these rare RET/PTC types have been found in papillary carcinomas from patients with a history of either environmental (after Chernobyl) or therapeutic exposure to ionizing radiation (Klugbauer et al. 1998, 2000, Klugbauer & Rabes 1999, Corvi et al. 2000, Salassidis et al. 2000, Saenko et al. 2003). The one exception is the ELKS-RET fusion identified in a case of papillary carcinoma with no apparent history of radiation exposure (Nakata et al. 1999). In the present study, we identify yet another novel RET/PTC rearrangement in a patient with no radiation exposure history.

It has been established that different RET partner genes involved in RET/PTC rearrangement share several common characteristics important for the oncogenic function of the chimeric gene. They are all expressed in thyroid follicular cells, providing an active promoter for the fusion gene. In addition, they all encode putative dimerization domains, either a coiled–coil or leucine zipper, essential for dimerization and ligand-independent activation of the RET (TK; Tong et al. 1997, Jhiang 2000). As for the RET gene, virtually all breakpoints occur within its 1.8 kb intron 11, leaving intact the TK domain of the receptor and therefore enabling the RET/PTC oncoprotein to bind SHC via Y1062 and activate the RAS–RAF–MAPK pathway in thyroid cells (Knauf et al. 2003).

All of these structural features can be found in the HOOK3-RET chimeric gene. HOOK3 encodes a protein belonging to a recently identified family of human cytosolic coiled-coil proteins that link cytoplasmic organelles to microtubules (Walenta et al. 2001). The three known human HOOK proteins, HOOK1, HOOK2, and HOOK3, contain conserved N-terminal domains which attach to microtubules, and more variable C-terminal domains which mediate binding to different organelles. The C-terminal portion of HOOK3 binds to Golgi membranes in vitro, suggesting that it may participate in defining the organization and localization of the mammalian Golgi complex within the cell (Walenta et al. 2001). Similar to all other family members, the HOOK3 protein contains an extended central coiled-coil motif, which was shown to mediate homodimerization in the Drosophila Hook protein (Kramer & Phistry 1996, Sevrioukov et al. 1999). In the HOOK3-RET fusion, the first 11 exons of the HOOK3 gene are fused to exons 12–20 of the RET gene. Two large coiled-coil domains of HOOK3 encoded by exons 7–11 (aa. 171–226 and 244–359) are preserved in the HOOK3-RET protein, indicating that it possesses a dimerization domain found in all known RET fusion partners. In the RET gene, the breakpoint is located within intron 11, similar to most other rearrangement types. Finally, we confirmed that the wild-type HOOK3 gene is expressed in thyroid follicular cells, providing an active promoter to drive the expression of the TK domain of RET. These qualities are expected to enable the HOOK3-RET with oncogenic properties, which indeed were identified in both in vitro and in vivo experiments.

In summary, we report a novel RET/PTC rearrangement resulting from RET fusion to the HOOK3 gene which leads to the formation of an oncogene.

	Acknowledgements

 
This study was supported by NIH grant R01 CA88041. Raffaele Ciampi is a recipient of the FIRC (Italian Federation for Cancer Research) fellowship. The authors declare that there is no conflict of interest that would prejudice the impartiality of this scientific work.

	References

Top Abstract Introduction Materials and methods Results Discussion References

 
Airaksinen MS, Titievsky A & Saarma M 1999 GDNF family neurotrophic factor signaling: four masters, one servant? Molecular and Cellular Neurosciences 13 313–325.[CrossRef][ISI][Medline]

Argani P, Zakowski MF, Klimstra DS, Rosai J & Ladanyi M 1998 Detection of the SYT-SSX chimeric RNA of synovial sarcoma in paraffin-embedded tissue and its application in problematic cases. Modern Pathology 11 65–71.[ISI][Medline]

Bongarzone I, Monzini N, Borrello MG, Carcano C, Ferraresi G, Arighi E, Mondellini P, Della Porta G & Pierotti MA 1993 Molecular characterization of a thyroid tumor-specific transforming sequence formed by the fusion of ret tyrosine kinase and the regulatory subunit RI alpha of cyclic AMP-dependent protein kinase A. Molecular and Cellular Biology 13 358–366.[Abstract/Free Full Text]

Bongarzone I, Butti MG, Coronelli S, Borrello MG, Santoro M, Mondellini P, Pilotti S, Fusco A, Della Porta G & Pierotti MA 1994 Frequent activation of ret proto-oncogene by fusion with a new activating gene in papillary thyroid carcinomas. Cancer Research 54 2979–2985.[Abstract/Free Full Text]

Ciampi R & Nikiforov YE 2005 Alterations of the BRAF gene in thyroid tumors. Endocrine Pathology 16 163–172.[CrossRef][ISI][Medline]

Ciampi R, Knauf JA, Kerler R, Gandhi M, Zhu Z, Nikiforova MN, Rabes HM, Fagin JA & Nikiforov YE 2005 Oncogenic AKAP9-BRAF fusion is a novel mechanism of MAPK pathway activation in thyroid cancer. Journal of Clinical Investigation 115 94–101.[CrossRef][ISI][Medline]

Corvi R, Berger N, Balczon R & Romeo G 2000 RET/PCM-1: a novel fusion gene in papillary thyroid carcinoma. Oncogene 19 4236–4242.[CrossRef][ISI][Medline]

De Micco C 2003 Ras mutations in follicular variant of papillary thyroid carcinoma. American Journal of Clinical Pathology 120 803–804.[ISI][Medline]

Frattini M, Ferrario C, Bressan P, Balestra D, De Cecco L, Mondellini P, Bongarzone I, Collini P, Gariboldi M, Pilotti S et al. 2004 Alternative mutations of BRAF, RET and NTRK1 are associated with similar but distinct gene expression patterns in papillary thyroid cancer. Oncogene 23 7436–7440.[CrossRef][ISI][Medline]

Fugazzola L, Pierotti MA, Vigano E, Pacini F, Vorontsova TV & Bongarzone I 1996 Molecular and biochemical analysis of RET/PTC4, a novel oncogenic rearrangement between RET and ELE1 genes, in a post-Chernobyl papillary thyroid cancer. Oncogene 13 1093–1097.[ISI][Medline]

Fusco A, Grieco M, Santoro M, Berlingieri MT, Pilotti S, Pierotti MA, Della Porta G & Vecchio G 1987 A new oncogene in human thyroid papillary carcinomas and their lymph-nodal metastases. Nature 328 170–172.[CrossRef][Medline]

Giordano TJ, Kuick R, Thomas DG, Misek DE, Vinco M, Sanders D, Zhu Z, Ciampi R, Roh M, Shedden K et al. 2005 Molecular classification of papillary thyroid carcinoma: distinct BRAF, RAS, and RET/PTC mutation-specific gene expression profiles discovered by DNA microarray analysis. Oncogene 24 6646–6656.[CrossRef][ISI][Medline]

Grieco M, Santoro M, Berlingieri MT, Melillo RM, Donghi R, Bongarzone I, Pierotti MA, Della Porta G, Fusco A & Vecchio G 1990 PTC is a novel rearranged form of the ret proto-oncogene and is frequently detected in vivo in human thyroid papillary carcinomas. Cell 60 557–563.[CrossRef][ISI][Medline]

Hundahl SA, Cady B, Cunningham MP, Mazzaferri E, McKee RF, Rosai J, Shah JP, Fremgen AM, Stewart AK & Holzer S 2000 Initial results from a prospective cohort study of 5583 cases of thyroid carcinoma treated in the United States during 1996. US and German Thyroid Cancer Study Group. An American College of Surgeons Commission on Cancer Patient Care Evaluation study. Cancer 89 202–217.[CrossRef][ISI][Medline]

Ishizaka Y, Ushijima T, Sugimura T & Nagao M 1990 cDNA cloning and characterization of ret activated in a human papillary thyroid carcinoma cell line. Biochemical and Biophysical Research Communications 168 402–408.[CrossRef][ISI][Medline]

Jhiang SM 2000 The RET proto-oncogene in human cancers. Oncogene 19 5590–5597.[CrossRef][ISI][Medline]

Kimura ET, Nikiforova MN, Zhu Z, Knauf JA, Nikiforov YE & Fagin JA 2003 High prevalence of BRAF mutations in thyroid cancer: genetic evidence for constitutive activation of the RET/PTC-RAS-BRAF signaling pathway in papillary thyroid carcinoma. Cancer Research 63 1454–1457.[Abstract/Free Full Text]

Klugbauer S & Rabes HM 1999 The transcription coactivator HTIF1 and a related protein are fused to the RET receptor tyrosine kinase in childhood papillary thyroid carcinomas. Oncogene 18 4388–4393.[CrossRef][ISI][Medline]

Klugbauer S, Demidchik EP, Lengfelder E & Rabes HM 1998 Detection of a novel type of RET rearrangement (PTC5) in thyroid carcinomas after Chernobyl and analysis of the involved RET-fused gene RFG5. Cancer Research 58 198–203.[Abstract/Free Full Text]

Klugbauer S, Jauch A, Lengfelder E, Demidchik E & Rabes HM 2000 A novel type of RET rearrangement (PTC8) in childhood papillary thyroid carcinomas and characterization of the involved gene (RFG8). Cancer Research 60 7028–7032.[Abstract/Free Full Text]

Knauf JA, Kuroda H, Basu S & Fagin JA 2003 RET/PTC-induced dedifferentiation of thyroid cells is mediated through Y1062 signaling through SHC-RAS-MAP kinase. Oncogene 22 4406–4412.[CrossRef][ISI][Medline]

Kramer H & Phistry M 1996 Mutations in the Drosophila hook gene inhibit endocytosis of the boss transmembrane ligand into multivesicular bodies. Journal of Cell Biology 133 1205–1215.[Abstract/Free Full Text]

Kunzli BM, Berberat PO, Zhu ZW, Martignoni M, Kleeff J, Tempia-Caliera AA, Fukuda M, Zimmermann A, Friess H & Buchler MW 2002 Influences of the lysosomal associated membrane proteins (Lamp-1, Lamp-2) and Mac-2 binding protein (Mac-2-BP) on the prognosis of pancreatic carcinoma. Cancer 94 228–239.[CrossRef][ISI][Medline]

Minoletti F, Butti MG, Coronelli S, Miozzo M, Sozzi G, Pilotti S, Tunnacliffe A, Pierotti MA & Bongarzone I 1994 The two genes generating RET/PTC3 are localized in chromosomal band 10q11.2. Genes Chromosomes Cancer 11 51–57.[ISI][Medline]

Nakata T, Kitamura Y, Shimizu K, Tanaka S, Fujimori M, Yokoyama S, Ito K & Emi M 1999 Fusion of a novel gene, ELKS, to RET due to translocation t(10;12)(q11;p13) in a papillary thyroid carcinoma. Genes Chromosomes Cancer 25 97–103.[CrossRef][ISI][Medline]

Pierotti MA, Santoro M, Jenkins RB, Sozzi G, Bongarzone I, Grieco M, Monzini N, Miozzo M, Herrmann MA, Fusco A et al. 1992 Characterization of an inversion on the long arm of chromosome 10 juxtaposing D10S170 and RET and creating the oncogenic sequence RET/PTC. PNAS 89 1616–1620.[Abstract/Free Full Text]

Saenko V, Rogounovitch T, Shimizu-Yoshida Y, Abrosimov A, Lushnikov E, Roumiantsev P, Matsumoto N, Nakashima M, Meirmanov S, Ohtsuru A et al. 2003 Novel tumorigenic rearrangement, Delta RFP/RET, in a papillary thyroid carcinoma from externally irradiated patient. Mutation Research 527 81–90.[ISI][Medline]

Salassidis K, Bruch J, Zitzelsberger H, Lengfelder E, Kellerer AM & Bauchinger M 2000 Translocation t(10;14)(q11.2:q22.1) fusing the kinetin to the RET gene creates a novel rearranged form (PTC8) of the RET proto-oncogene in radiation-induced childhood papillary thyroid carcinoma. Cancer Research 60 2786–2789.[Abstract/Free Full Text]

Santoro M, Dathan NA, Berlingieri MT, Bongarzone I, Paulin C, Grieco M, Pierotti MA, Vecchio G & Fusco A 1994 Molecular characterization of RET/PTC3; a novel rearranged version of the RET proto-oncogene in a human thyroid papillary carcinoma. Oncogene 9 509–516.[ISI][Medline]

Sevrioukov EA, He JP, Moghrabi N, Sunio A & Kramer H 1999 A role for the deep orange and carnation eye color genes in lysosomal delivery in Drosophila. Molecular Cell 4 479–486.[CrossRef][ISI][Medline]

Soares P, Trovisco V, Rocha AS, Lima J, Castro P, Preto A, Maximo V, Botelho T, Seruca R & Sobrinho-Simoes M 2003 BRAF mutations and RET/PTC rearrangements are alternative events in the etiopathogenesis of PTC. Oncogene 22 4578–4580.[CrossRef][ISI][Medline]

Takahashi M 1988 Structure and expression of the ret transforming gene. IARC Scientific Publications 189–197.

Takahashi M, Ritz J & Cooper GM 1985 Activation of a novel human transforming gene, ret, by DNA rearrangement. Cell 42 581–588.[CrossRef][ISI][Medline]

Tomlins SA, Rhodes DR, Perner S, Dhanasekaran SM, Mehra R, Sun XW, Varambally S, Cao X, Tchinda J, Kuefer R et al. 2005 Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science 310 644–648.[Abstract/Free Full Text]

Tong Q, Xing S & Jhiang SM 1997 Leucine zipper-mediated dimerization is essential for the PTC1 oncogenic activity. Journal of Biological Chemistry 272 9043–9047.[Abstract/Free Full Text]

Walenta JH, Didier AJ, Liu X & Kramer H 2001 The Golgi-associated hook3 protein is a member of a novel family of microtubule-binding proteins. Journal of Cell Biology 152 923–934.[Abstract/Free Full Text]

Xing M 2005 BRAF mutation in thyroid cancer. Endocrine-Related Cancer 12 245–262.[Abstract/Free Full Text]

Zhu Z, Gandhi M, Nikiforova MN, Fischer AH & Nikiforov YE 2003 Molecular profile and clinical-pathologic features of the follicular variant of papillary thyroid carcinoma. An unusually high prevalence of ras mutations. American Journal of Clinical Pathology 120 71–77.[CrossRef][ISI][Medline]

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