A novel multicolor hybridization scheme applied to localization of a transcribed sequence (D10S170/H4) and deletion mapping in the thyroid cancer cell line TPC-1

Jossart, Gregg H.; O’Brien, Benjamin; Cheng, Jan‐Fang; Tong, Qiang; Jhiang, Sissy; Duh, Quan Yang; Clark, Orlo H.; Weier, Heinz Ulrich G.

doi:10.1159/000134495

Cited by 32 publications

(33 citation statements)

References 5 publications

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“…A group of pathologists have reported that 90% of RET/PTC1 positive thyroid tumours fail to express the unrearranged allele of H4(D10S170) (Sheils et al, 2000). The same phenomenon has been observed in the TPC-1 thyroid papillary carcinoma cell line, a line harbouring RET/PTC1 oncogene activation that has been reported to have a gene deletion encompassing the normal H4(D10S170) allele (Jossart et al, 1996). Taken together, these data suggest that the presence of the unrearranged normal H4(D10S170) gene product could exert a negative action on cell proliferation and that a growth advantage might result from the loss of the normal allele of this gene, likely during tumour progression.…”

Section: Discussionmentioning

confidence: 71%

“…The same results ( Figure 2a) were obtained in a human thyroid papillary carcinoma cell line, established from a tumour carrying the RET/PTC1 oncogene activation, named TPC-1. It has been shown that these cells have lost by deletion the wt H4(D10S170) allele not involved in the rearrangement with RET (Jossart et al, 1996).…”

Section: H4 Is a Nuclear And Cytosolic Proteinmentioning

confidence: 99%

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H4(D10S170), a gene frequently rearranged with RET in papillary thyroid carcinomas: functional characterization

et al. 2004

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Human thyroid papillary carcinomas are characterized by rearrangements of the RET protooncogene with a number of heterologous genes, which generate the RET/papillary thyroid carcinoma (PTC) oncogenes. One of the most frequent variants of these recombination events is the fusion of the intracellular kinase-encoding domain of RET to the first 101 amino acids of a gene named H4(D10S170). We have characterized the H4(D10S170) gene product, showing that it is a ubiquitously expressed 55 KDa nuclear and cytosolic protein that is phosphorylated following serum stimulation. This phosphorylation was found to depend on mitogen-activated protein kinase (MAPK) Erk1/2 activity and to be associated to the relocation of H4(D10S170) from the nucleus to the cytosol. Overexpression of the H4(D10S170) gene was able to induce apoptosis of thyroid follicular epithelial cells; conversely a carboxy-terminal truncated H4(D10S170) mutant H4(1-101), corresponding to the portion included in the RET/PTC1 oncoprotein, behaved as dominant negative on the proapoptotic function and nuclear localization of H4(D10S170). Furthermore, conditional expression of the H4(D10S170)-dominant negative truncated mutant protected cells from stress-induced apoptosis. The substitution of serine 244 with alanine abrogated the apoptotic function of H4(D10S170). These data suggest that loss of the H4(D10S170) gene function might have a role in thyroid carcinogenesis by impairing apoptosis.

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Section: Discussionmentioning

confidence: 71%

Section: H4 Is a Nuclear And Cytosolic Proteinmentioning

confidence: 99%

H4(D10S170), a gene frequently rearranged with RET in papillary thyroid carcinomas: functional characterization

et al. 2004

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“…Transient expression of H4(D10S170) gene product was associated with induction of apoptosis in thyroid cells. TPC-1 cells, which harbours the RET/PTC1 rearrangement and have lost the expression of the normal unrearranged H4(D10S170) allele (Jossart et al, 1996), transiently transfected with H4(D10S170)wt, showed impaired apoptosis if pretreated with 50 nM wortmannin, 1 mM caffeine and 10 mM KU55933 (Hickson et al, 2004), known inhibitors of ATM kinase activity (Figure 5a). In addition, in TPC-1 cells, H4 T434A expression had a dominant negative effect on H4wt-mediated apoptosis (Figure 5a).…”

Section: H4(d10s170) Location and Phosphorylation In A-t Cellsmentioning

confidence: 99%

Involvement of H4(D10S170) protein in ATM-dependent response to DNA damage

et al. 2007

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H4(D10S170) gene has been identified upon its frequent rearrangement with RET in papillary thyroid tumours (RET/PTC1). The kinase ataxia telangectasia mutated (ATM) phosphorylates a limited number of downstream protein targets in response to DNA damage. We investigated the potential role of H4(D10S170) in DNA damage signaling pathways. We found that in cells treated with etoposide or ionizing radiation (IR), H4(D10S170) underwent ATM-mediated phosphorylation at Thr 434, stabilizing nuclear H4. In ataxia telangectasia cells (A-T), endogenous H4(D10S170) was localized to cytoplasm and was excluded from the nucleus. Moreover, H4(D10S170) was not phosphorylated in ATM-deficient lymphoblasts after ionizing irradiation. Inhibition of ATM kinase interfered with H4(D10S170) apoptotic activity, and expression of H4 with threonine 434 mutated in Alanine, H4 T434A, protected the cells from genotoxic stress-induced apoptosis. Most importantly, after exposure to IR we found that silencing of H4(D10S170) in mammalian cells increased cell survival, as shown by clonogenic assay, allows for DNA synthesis as evaluated by bromodeoxyuridine incorporation and permits cells to progress into mitosis as demonstrated by phosphorylation on Histone H3. Our results suggest that H4(D10S170) is involved in cellular response to DNA damage ATM-mediated, and that the impairment of H4(D10S170) gene function might have a role in thyroid carcinogenesis.

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“…The order of these probes on 10q is reported elsewhere. 32 The RET probe was labeled with SpectrumGreen™-dUTP and the H4 probe with SpectrumRed™-dUTP (Vysis, Inc., Richmond, UK) using the Nick Translation Kit (Vysis, Inc.). A standard hybridization method was used according to the manufacturer's protocol.…”

Section: Fluorescence In Situ Hybridization (Fish) Analysismentioning

confidence: 99%

“…This suggests that H4 may be a tumor suppressor gene. 32 We evaluated H4 gene expression in FB-2 cells by using a semiquantitative RT-PCR and primers encompassing the 3Ј half of the gene downstream from the rearrangement. Like TPC-1 cells, FB-2 cells did not express the H4 gene (Fig.…”

Section: Ret Rearrangement In Fb-2 Cellsmentioning

confidence: 99%

Establishment of a non‐tumorigenic papillary thyroid cell line (FB‐2) carrying the RET/PTC1 rearrangement

Basolo

Giannini

Toniolo

et al. 2001

Intl Journal of Cancer

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A novel human thyroid papillary carcinoma cell line (FB-2) has been established and characterized. FB-2 cells harbor the RET/PTC1 chimeric oncogene in which the RET kinase domain is fused to the H4 gene. FB-2 cells neither formed colonies in semisolid media nor induced tumors after heterotransplant into severe combined immunodeficient mice. However, HMGI(Y), HMGI-C and c-myc genes, which are associated to thyroid cell transformation, were abundantly expressed in FB-2 cells but not in normal thyroid cells. FB-2 cells only partially retained the differentiated thyroid phenotype. In fact, the PAX-8 gene, which codes for a transcriptional factor required for thyroid cell differentiation, was expressed, while thyroglobulin, TSH-receptor and thyroperoxidase genes were not. Moreover, FB-2 cells produced high levels of interleukin (IL)-6 and IL-8. Key words: thyroid; papillary carcinoma; cell line; RET rearrangementTumors of follicular thyroid cells are highly heterogeneous in terms of histology and response to treatment. Malignant thyroid tumors include (i) well-differentiated carcinomas, which comprise follicular (FTC) and papillary (PTC) carcinomas; (ii) poorly differentiated carcinomas; and (iii) undifferentiated carcinomas. 1 Well-differentiated PTC is the most frequent type of thyroid cancer and it accounts for the vast majority of thyroid carcinomas associated with previous exposure to ionizing radiation. 2 The prognosis of papillary thyroid carcinoma is generally favorable. However, a number of patients develop recurrences (local or nodal) and distant metastases, or die. 3 Rearrangements of the RET proto-oncogene represent the most frequent genetic lesion found in thyroid papillary carcinomas. 4 These rearrangements cause the fusion of the tyrosine kinase encoding domain of the RET gene to heterologous genes and lead to the generation of the chimeric RET/PTC oncogenes. Several RET/PTC oncogenes have been identified that differ in the fusion partner. 5-11 RET/PTC1 and RET/PTC3 are the most prevalent RET/PTC variants. 11 In RET/PTC1 and RET/PTC 3, the fusion occurs with the H4 (or D10S170) gene 5 and the RFG gene, respectively. 7 A chromosomal inversion [inv (10) (q11.2q21)] accounts for the generation of RET/PTC1, whereas a cytogenetically undetectable paracentric inversion within 10q11.2 accounts for RET/PTC 3 activation. 11 All the genes fused to RET are ubiquitously expressed and therefore able to drive the expression of truncated forms of RET in thyroid follicular cells that normally do not express it. The RET fusion partners encode dimerizationmotif-containing proteins, which mediate constitutive activation of the rearranged RET kinase, phosphorylation of RET on tyrosine residues and chronic stimulation of RET signal transduction. 12 RET/PTC oncogenes play a pivotal role in the pathogenesis of human thyroid papillary carcinomas. Indeed they transform thyroid cells in culture, 13 and transgenic mice carrying the RET/PTC1and RET/PTC3 oncogene, under the transcriptional control of the thyroid-specific rat thyr...

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A novel multicolor hybridization scheme applied to localization of a transcribed sequence (D10S170/H4) and deletion mapping in the thyroid cancer cell line TPC-1

Cited by 32 publications

References 5 publications

H4(D10S170), a gene frequently rearranged with RET in papillary thyroid carcinomas: functional characterization

H4(D10S170), a gene frequently rearranged with RET in papillary thyroid carcinomas: functional characterization

Involvement of H4(D10S170) protein in ATM-dependent response to DNA damage

Establishment of a non‐tumorigenic papillary thyroid cell line (FB‐2) carrying the RET/PTC1 rearrangement

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