Centrosomal and mitotic abnormalities in cell lines derived from papillary thyroid cancer harboring specific gene alterations

Molecular Cytogenetics

Roberta Vanni¹  Paolo Degan²  Daniela V Frau¹  Paola Caria¹  Silvia Viaggi²  Irena Maric² 

[1] Dipartimento di Scienze e Tecnologie Biomediche, Università di Cagliari, 09042, Italy;IRCCS Azienda Ospedaliera Universitaria San Martino - IST - Istituto Nazionale per la Ricerca sul Cancro, Genova, 16132, Italy

关键词: BRAF;    RET/PTC;    mitotic spindle;    centrosome;    thyroid carcinoma;   

Others  :  1151940
DOI  :  10.1186/1755-8166-4-26

 received in 2011-09-11, accepted in 2011-11-16,  发布年份 2011

【 摘 要 】

Background

Differentiated thyroid carcinoma offers a good model to investigate the possible correlation between specific gene mutations and chromosome instability. Papillary thyroid neoplasms are characterized by different mutually exclusive genetic alterations, some of which are associated with aneuploidy and aggressive phenotype.

Results

We investigated the centrosome status and mitotic abnormalities in three thyroid carcinoma-derived cell lines, each maintaining the specific, biologically relevant gene alteration harbored by the parental tumors: RET/PTC1 rearrangement in TPC1; heterozygous and homozygous BRAF^V600E mutation in K1 and in B-CPAP, respectively. B-CPAP cells showed a statistically significant (P < 0.01) higher frequency of abnormal mitotic figures compared to TPC1 and K1 cells.

Conclusions

Our data indicate that RET/PTC1 oncogenic activity is not related to mitotic chromosome impairment and missegregation whereas, based on the consistent difference in types/frequencies of centrosome and spindle abnormalities observed between K1 and B-CPAP cells, the hetero/homozygous allelic status of BRAF^V600E mutation seems to be not irrelevant in respect to chromosomal instability development.

【 授权许可】

   
2011 Maric et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Ricke RM, van Ree JH, van Deursen JM: Whole chromosome instability and cancer: a complex relationship. Trends Genet 2008, 24:457-466.
• [2]King RW: When 2+2 = 5: the origins and fates of aneuploid and tetraploid cells. Biochim Biophys Acta 2008, 1786:4-14.
• [3]McPherson JP, Hande MP, Poonepalli A, Lemmers B, Zablocki E, Migon E, Shehabeldin A, Porras A, Karaskova J, Vukovic B, Squire J, Hakem R: A role for Brca1 in chromosome end maintenance. Hum Mol Genet 2006, 15:831-838.
• [4]Fukasawa K: Oncogenes and tumour suppressors take on centrosomes. Nat Rev Cancer 2007, 7:911-924.
• [5]Brinkley BR: Managing the centrosome numbers game: from chaos to stability in cancer cell division. Trends Cell Biol 2001, 11:18-21.
• [6]Nikiforov YE, Nikiforova MN: Molecular genetics and diagnosis of thyroid cancer. Nat Rev Endocrinol 2011, 7:569-80.
• [7]Meireles AM, Preto A, Rocha AS, Rebocho AP, Máximo V, Pereira-Castro I, Moreira S, Feijão T, Botelho T, Marques R, Trovisco V, Alves C, Velho S, Soares P, Sobrinho-Simões M: Molecular and genotypic characterization of human thyroid follicular cell carcinoma-derived cell lines. Thyroid 2007, 17:707-715.
• [8]Mitelman F, Johansson B, Mertens F, (Eds): Mitelman Database of Chromosome Aberrations and Gene Fusions in Cancer. [http://cgap.nci.nih.gov/Chromosomes/Mitelman] webcite 2011. accession 15 October 2011
• [9]Rodrigues R, Roque L, Espadinha C, Pinto A, Domingues R, Dinis J, Catarino A, Pereira T, Leite V: Comparative genomic hybridization, BRAF, RAS, RET, and oligo-array analysis in aneuploid papillary thyroid carcinoma. Oncol Rep 2007, 18:917-26.
• [10]Mitsutake N, Knauf JA, Mitsutake S, Mesa C Jr, Zhang L, Fagin JA: Conditional BRAFV600E expression induces DNA synthesis, apoptosis, dedifferentiation, and chromosomal instability in thyroid PCCL3 cells. Cancer Res 2005, 65:2465-73.
• [11]Niu D, Murata S, Kondo T, Nakazawa T, Kawasaki T, Ma D, Yamane T, Nakamura N, Katoh R: Involvement of centrosomes in nuclear irregularity of thyroid carcinoma cells. Virchows Arch 2009, 455:149-157.
• [12]Ishizaka Y, Ushijima T, Sugimura T, Nagao M: cDNA cloning and characterization of ret activated in a human papillary thyroid carcinoma cell line. Biochem Biophys Res Commun 1990, 168:402-408.
• [13]Lui WO, Zeng L, Rehrmann V, Deshpande S, Tretiakova M, Kaplan EL, Leibiger I, Leibiger B, Enberg U Höög A, Larsson C, Kroll TG: CREB3L2-PPARgamma fusion mutation identifies a thyroid signaling pathway regulated by intramembrane proteolysis. Cancer Res 2008, 68:7156-7164.
• [14]Caria P, Vanni R: Cytogenetic and molecular events in adenoma and well-differentiated thyroid follicular-cell neoplasia. Cancer Genet Cytogenet 2010, 203:21-29.
• [15]van Staveren WC, Solís DY, Hébrant A, Detours V, Dumont JE, Maenhaut C: Human cancer cell lines: Experimental models for cancer cells in situ? For cancer stem cells? Biochim Biophys Acta 2009, 1795:92-103.
• [16]Mahoney NM, Goshima G, Douglass AD, Vale RD: Making microtubules and mitotic spindles in cells without functional centrosomes. Curr Biol 2006, 16:564-569,.
• [17]Cui Y, Borysova MK, Johnson JO, Guadagno TM: Oncogenic B-Raf(V600E) induces spindle abnormalities, supernumerary centrosomes, and aneuploidy in human melanocytic cells. Cancer Res 2010, 15:675-684.
• [18]Davies H, Bignell G R, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W, Davis N, Dicks E, Ewing R, Floyd Y, Gray K, Hall S, Hawes R, Hughes J, Kosmidou V, Menzies A, Mould C, Parker A, Stevens C, Watt S, Hooper S, Wilson R, Jayatilake H, Gusterson BA, Cooper C, Shipley J, et al.: Mutations of the BRAF gene in human cancer. Nature 2002, 417:949-954.
• [19]Rubinstein JC, Sznol M, Pavlick AC, Ariyan S, Cheng E, Bacchiocchi A, Kluger HM, Narayan D, Halaban R: Incidence of the V600K mutation among melanoma patients with BRAF mutations, and potential therapeutic response to the specific BRAF inhibitor PLX4032. J Transl Med 2010, 14(8):67.
• [20]Fugazzola L, Puxeddu E, Avenia N, Romei C, Cirello V, Cavaliere A, Faviana P, Mannavola D, Moretti S, Rossi S, Sculli M, Bottici V, Beck-Peccoz P, Pacini F, Pinchera A, Santeusanio F, Elisei R: Correlation between B-RAFV600E mutation and clinico-pathologic parameters in papillary thyroid carcinoma: data from a multicentric Italian study and review of the literature. Endocr Relat Cancer 2006, 13:455-64.
• [21]Shinmura K, Bennett RA, Tarapore P, Fukasawa K: Direct evidence for the role of centrosomally localized p53 in the regulation of centrosome duplication. Oncogene 2007, 26:2939-44.
• [22]Rhoden KJ Unger K, Salvatore G, Yilmaz Y, Vovk V, Chiappetta G, Qumsiyeh MB, Rothstein JL, Fusco A, Santoro M, Zitzelsberger H, Tallini G: RET/Papillary Thyroid Cancer Rearrangement in Nonneoplastic Thyrocytes: Follicular Cells of Hashimoto's Thyroiditis Share Low-Level Recombination Events with a Subset of Papillary Carcinoma. J Clin Endocrinol Metab 2006, 91:2414-2423.
• [23]Health Protection Agency Culture Collections [http://www.hpa.org.uk] webcite
• [24]Dettori T, Frau DV, Garcia JL, Pierantoni G, Lee C, Hernandez JM, Fusco A, Morton CC, Vanni R: Comprehensive conventional and molecular cytogenetic characterization of BCPAP, a human papillary thyroid carcinoma-derived cell line. Cancer Genet Cytogenet 2004, 151:171-177.
• [25]Ribeiro FR, Meireles AM, Rocha AS, Teixeira MR: Conventional and molecular cytogenetics of human non-medullary thyroid carcinoma: characterization of eight cell line models and review of the literature on clinical samples. BMC Cancer 2008, 16:371.
• [26]Liu R, Liu D, Trink E, Bojdani E, Ning G, Xing M: The Akt-specific inhibitor MK2206 selectively inhibits thyroid cancer cells harboring mutations that can activate the PI3K/Akt pathway. J Clin Endocrinol Metab 2011, 96:E577-85.
• [27]Lemoine NR, Mayall ES, Jones T, Sheer D, McDermid S, Kendall-Taylor P, Wynford-Thomas D: Characterisation of human thyroid epithelial cells immortalised in vitro by simian virus 40 DNA transfection. Br J Cancer 1989, 60:897-903.
• [28]Krämer A, Schweizer S, Neben K, Giesecke C, Kalla J, Katzenberger T, Ho AD, Ott G: Centrosome aberrations as a possible mechanism for chromosomal instability in non-Hodgkin's lymphoma. Leukemia 2003, 17:2207-2213.