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Department of Molecular Biology, Institute of Gerontology, Nippon Medical School, 1-396, Kosugi-cho, Nakahara-ku, Kawasaki 211-8533, Japan
1 Kanagawa Prefectural Cancer Center, 1-1-2, Nakao, Asahi-ku, Yokohama 241-0815, Japan
2 Department of Surgery, Nippon Medical School and
3 Ito Hospital, 4-3-6, Jinguumae, Shibuya-ku, Tokyo 150-8308, Japan
4 Laboratory for Medical Informatics, RIKEN, 22-7-1, Suehiro-cho, Tsurumi-ku, Kanagawa 230-0045, Japan
5 Laboratory of Molecular Medicine, Human Genome Center, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan
(Requests for offprints should be addressed to M Emi; Email: memi{at}nms.ac.jp)
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Abstract |
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Introduction |
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TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
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The clinical behavior of ATC is markedly distinct from other types of thyroid cancer. It is one of the most virulent cancers of all human malignancies (Sherman 2003), with a mean survival time among patients of less than 1 year after diagnosis, regardless of treatment (Passler et al. 1999, Voutilainen et al. 1999). Differences in biological characteristics among thyroid tumors might be explained by variations in the pattern of sequential somatic mutations among genes that participate in the mechanisms of growth and differentiation. Although mutation of TP53 (Kitamura et al. 2000) and ß-catenin (Garcia-Rostan et al. 1999) are observed in some ATCs, the former probably inactivating a tumor suppressor and the latter activating an oncogenic function, the underlying molecular mechanism involved in this type of thyroid cancer is poorly understood.
Using 11 anaplastic cancer cell lines (ACLs) and ten primary ATCs, we investigated gene-expression profiles on a cDNA microarray consisting of 25 344 genes. The tumors displayed remarkably characteristic profiles that should be useful for molecular diagnosis, for prediction of prognosis and for identifying potential target molecules for novel drugs to treat this type of cancer.
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Materials and methods |
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TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
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Eleven cell lines that were derived from ATCs, 8305c, 8505c, ARO, FRO, TTA1, TTA2, TTA3, KTA1, KTA2, KTA3 and KTA4, were used for this study. These cell lines were maintained with Dulbecco’s modified Eagle’s medium (Invitrogen, Carlsbad, CA, USA) for 8305c and 8505c, minimum essential medium for ARO and FRO and RPMI 1640 for the other seven lines. All media contained 10% fetal bovine serum but no antibiotics. The cells were cultured in a 37°C incubator under 5% CO2 atmosphere.
Patients and specimens
Primary ATC and non-cancerous thyroid tissues were excised from ten patients who underwent surgery at the Ito Hospital, Tokyo, Japan; the samples were frozen immediately and stored at –80°C. All patients had given informed consent according to guidelines approved by the Institutional Research Board. All tumor specimens that we analyzed contained more than 70% tumor cells. To equalize the tumor-associated deviation in gene expression from normal thyroid, an equal amount of a mixture of RNAs from the ten non-cancerous tissues was used as a normal control for competitive hybridizations on the microarray.
RNA extraction and RNA amplification
Each tissue was homogenized with TRIZOL reagent (Invitrogen) according to the manufacturer’s instructions for RNA extraction. One microgram of extracted RNA was electrophoresed on a 3.0% formaldehyde denaturing gel to eliminate degenerated RNA. Samples with 28S/18S ratios greater than 1.5 were selected for subsequent purification using RNeasy kits (QIAGEN, Valencia, CA, USA) to eliminate contamination with DNA. Total RNA was prepared for microarray analysis using T7 RNA polymerase-based amplification with MessageAmp aRNA kits (Ambion, Austin, TX, USA). In the first round, 5 µg aliquots of total RNA were used as templates for amplification, then 2µg aliquots of first-amplified RNA (aRNA) became the templates for second-round amplification. After the second-round amplification, aRNAs were purified with RNeasy purification kits, the amount of each aRNA was measured by a spectrophotometer, and its quality was checked by formaldehyde-agarose gel electrophoresis.
Preparation of cDNA microarray
The microarray contains 27 648 genes and expression sequence tags (ESTs) were generated with Spot Ready DNA for microarray (Amersham Biosciences Corp., Piscataway, NJ, USA). This cDNA panel was spotted using microarray spotter generation III (Amersham Biosciences Corp.) on microarray slide type 7 (Amersham Biosciences Corp.) and it was then cross-linked by u.v.
Labeling of aRNA and competitive hybridization
To generate hybridization probes, a 3 µg aliquot of each second-round aRNA was rendered fluorescent with the amino allyl cDNA labeling kit (Ambion), following the manufacturer’s protocol. Probes derived from cell lines and from the normal thyroid gland pool were labeled respectively with Cy5 or Cy3 mono-reactive dye (Amersham International plc, Amersham, Bucks, UK). To eliminate incorporated dye, the labeled probes were cleaned up with QIAquick PCR purification kits (Qiagen).
Fluorescent labeled probe (15 pmol) from each cell line was mixed with the Cy3-labeled normal control in 4xmicroarray hybridization buffer (Amersham Biosciences Corp.) and de-ionized formamide. The probe mixtures were hybridized for 12 h at 40°C, then washed once with 0.1xSSC, 0.2% SDS for 5 min and twice for 10 min in the same washing solution. All procedures were performed with an automated slide processor system (Amersham Biosciences Corp.) After hybridization, fluorescent signals were scanned with GenePix 4000 (Amersham International plc and data were collected by GenePix Pro 3.0 software (Amersham International plc). Scanned signals were normalized by a global method (Manos & Jones 2001, Yang et al. 2002).
Analysis of data
We performed random permutation tests to distinguish genes that were expressed differently between ACL and normal thyroid gland (Kitahara et al. 2002). The criteria for selection of discriminating genes were (1) signal/noise ratio of the gene greater than 3.0 in at least ten cell lines, (2) P value in a random permutation test lower than 0.0001 and (3) expression in cancer at least twofold stronger than normal (over-expressed) or half that of normal thyroid tissue (under-expressed). Genes that fitted these criteria were considered significant for discrimination.
Semi-quantitative RT-PCR for ACL and ATC
To confirm the microarray results we performed semi-quantitative RT-PCR (SQ-PCR) analysis of genes selected according to the following criteria: (1) the P value in permutation tests after microarray analysis was below 0.000001 and (2) expression in ACL was either threefold stronger or one-third weaker than in the normal thyroid gland. In addition to these selected genes, expression of p53 gene was evaluated in ATC samples because p53 is considered to be one of the key genes in ATC carcinogenesis. Five normal thyroid samples served as a control. cDNA was reverse transcribed from 10 µg of each total RNA in the usual manner. To adjust the amount of transcribed cDNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was selected as an internal control and SQ-PCR experiments were done as previously described, after adjustment of concentrations (Ono et al. 2000). The primer sequences for GAPDH were 5'-GGAAGGT GAAGGTCGGAGT-3'(forward) and 5'-TGGGTGG AATCATATTGGAA-3'(reverse).
Sequence information was collected from the NCBI GenBank (http://www.ncbi.nlm.nih.gov/), and all primers were designed with primer 3 software (http://www-genome.wi.mit.edu/cgi-bin/primer/primer3_www.cgi). Information about the PCR primers is available upon request to the corresponding author. SQ-PCR experiments were performed with 1 µl cDNA for the template, 5U Takara EX Taq (Takara, Otsu, Japan), 1xPCR buffer (10 mM Tris–HCl, 50 mM KCl and 1.5 mM MgCl2) and reverse primers in 30 µl of total reaction mixture. PCR conditions for each gene were optimized in their respective linear phases of amplification.
For evaluation of differences in gene expression between ACL/ATC and normal thyroid gland, 10 µl of each SQ-PCR product was electrophoresed on a 2.0% agarose gel and stained with ethidium bromide. After staining, the density of each sample spot was measured by AlphaImager 3300 (AlphaIonotech, San Leandro, CA, USA) with background revision. A 16 bit imaging score was acquired from each sample. All SQ-PCR experiments were duplicated. We applied Student’s t-tests to the results of the SQ-PCR assay; P values smaller than 0.05 were considered statistically significant. All statistical procedures were archived by Statview version 5.0 software (SAS Institute Inc., Cary, NC, USA).
Immunohistochemical analysis
To determine a correlation between RNA expression and protein expression, we performed immunohistochemical analyses using ten paraffin-embedded samples of primary ATC in three selected genes. GDI2 was significantly over-expressed with microarray analysis, stathmin and destrin were significantly over-expressed with microarray analysis and SQ-PCR in both ACL and ATC cases. All samples were collected at the Ito Hospital, Tokyo, Japan. Antibodies for this assay were available for GDI2 (1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA), Op-18 (stathmin) (1:100 dilution; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and destrin/ADF (1:100 dilution; Sigma, St Louis, MO, USA). Antigens were microwaved prior to immunostaining with VECTA-STAIN Elite ABC kits (Vector Laboratories Inc., Burlingame, CA, USA) and Dako ENVISION kits (Dako Corporation, Carpinteria, CA, USA) following the manufacturers’ instructions. The sections were counterstained with hematoxylin, and then scanned at low power to identify areas that were evenly stained. Estimates of the numbers of positive cells were scored as follows: negative, 0%; 1, 1–10%; 2, 11–25%; 3, 26–50%; 4, >50% positive (Saiz et al. 2002). Two independent investigators performed the estimation.
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Results |
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TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
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Permutation tests selected 31 genes and ESTs were up-regulated in ACL compared with normal thyroid gland. Those genes, along with their accession numbers, speculated function and chromosomal position are shown in Table 1
. Twenty-four of these genes have known or suggested functions. Among the over-expressed group were genes encoding small nuclear ribonucleoprotein, stathmin (Op-18) and DNA topoisomerase III, all apparently related to mechanisms of cell growth, were up-regulated in ACL. On the other hand, metabolism-related genes such as ATP5A1, ATP synthase and ODC1 were also over-expressed in ACL; however, these result might simply reflect the activated cell dynamics in the immortalized cells. In other categories, several genes encoding ribosomal protein were expressed dominantly in ACL, although a literature search revealed no implied connection between expression levels of those genes and thyroid cancer, especially in ATC, the anaplastic form in particular.
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The 56 genes that were under-expressed in ACL in comparison with the normal thyroid gland are listed in Table 2. Fifty-one of these were known genes, with a wide distribution of speculated functions. For example, ubiquitin-activating enzyme E1 is thought to be a tumor-suppressor gene, while the proto-oncogenes encoding c-fes/fps and human receptor protein tyrosine phosphatase hPTP-J precursor were unexpectedly under-expressed in ACL. These results suggested either that the proposed functions of the listed gene are variable, or the genes themselves were altered in ATC cell lines. Moreover, genes encoding laminin B2 chain and PLOD3, said to relate to cell structure, were down-regulated in ACL. A gene expression portrait of all 87 genes with altered expression in the cell lines is shown in Fig. 1.
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To validate the microarray results, we performed SQ-PCR experiments using samples from 11 cancer cell lines and ten primary ATC tumors, as well as tissues from five normal thyroid glands, and evaluated gene expression after normalization of signals according to the expression of GAPDH. The genes to be tested by SQ-PCR were chosen according to the criteria of (1) a P value below 0.000001 in the permutation test of microarray results and (2) expression levels in tumor at least threefold stronger or one-third weaker than that of normal thyroid gland tissue, as shown in Materials and methods. Figures 2 and 3 show SQ-PCR results for 12 of the over-expressed genes (Fig. 2A for ACL, Fig. 2B for ATC) and 15 under-expressed genes (Fig. 3A for ACL, (Fig. 3B for ATC). All of the results corroborated our microarray data, with statistical significance evaluated by Student’s t-test.
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subunit (IDH3G), C21orf97 and sex hormone-binding globulin (SHBG), were limited to cancer cell lines.
With regard to the p53 gene, the expression was slightly decreased (0.92-fold) in ACLs but it did not reach statistical significance with microarray analysis. We also tested p53 gene expression in ten ATC samples with SQ-PCR. The decreased expression was seen in ATC compared with normal thyroid tissue (P = 0.03) (Fig. 4).
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To evaluate the correlation between microarray profiling and protein expression, we performed immunohistochemical analysis of three antigenic proteins for which antibodies were commercially available (GDI2, stathmin and destrin). GDI2 was highly expressed (scored over 3) in seven of the ten primary ATC tumors. Stathmin (Op18) was also highly expressed in primary ATCs, with six tumors scoring over 3. Destrin was stained in five cases, mainly in cytoplasm. The representative results of these experiments are shown in (Fig. 5. On the other hand, all genes showed lower expression (staining) in part of the normal thyroid tissue. Table 3 summarizes the results of immunostaining analysis of GDI2, stathmin and destrin in all ten primary ATC cases. Consistent correlation between cDNA microarray data, SQ-PCR data and immunohistochemistry provides solid verification for the present study. In addition, this is the first report of over-expression in RNA and protein levels of these three genes in human thyroid malignancies as far as we can find in the literature.
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Discussion |
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TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
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For this study we did have access to 11 established ACLs for analysis on a cDNA microarray, which allowed us to construct a molecular portrait of ATC-derived cells. This is the first report to document a comprehensive gene expression profile of ATC. At present, p53 is the most well-known gene that might be responsible for ATC carcinogenesis (Kitamura et al. 2000); however, the expression level of p53 did not alter significantly between ACL and normal thyroid tissue with microarray analysis. In ATC samples, expression of p53 was weakly decreased. In addition, point mutations of p53 were found in five ACLs (TTA1: codon 72, CGC (wild type)-CTC (mutant); KTA2: codon 158, AAC-GAC; 8305c: codon 248, CGG-GGG; ARO: codon 273, CGT-CAT; KTA3: codon 276, GCC-CCC) All mutations were missense mutations but there was no specific mutation spectrum in ACLs. Hence it is difficult to explain the carcinogenic mechanism of only p53 single gene alterations. It is therefore necessary to examine gene expression alterations widely through the human genome to discover the novel genes responsible for ACL/ATC carcinogenesis.
Although some discrepancies are likely to exist between cell lines and primary ATCs, in general cell lines are thought to reflect, to a large degree, the characteristics of their tumors of origin; for example, a relationship between gene expression profiles of cell lines and primary tumor has been confirmed in bladder cancer (Sanchez-Carbayo et al. 2002). Our microarray analysis identified the up-regulation of 31 genes in the panel of 11 ACLs. Some of the over-expressed genes encoded ribosomal proteins, and several others encoded metabolism-related proteins such as lactate dehydrogenase A, B or ATP5A1 (ATP synthase); however, these may have been over-expressed simply as a result of active dynamism of the immortalized cells. Among the 31 genes in this list, GDI2 (located on 10p15) binds and solubilizes several membrane-associated Rab proteins in a GDP/GTP-dependent manner (Chinni et al. 1998). Amplification of chromosome 10p has been observed in a small number of head and neck cancers (Speicher et al. 1995). Our study showed up-regulation and over-expression of GDI2 in both ACL and ATC, suggesting that GDI2 contributes to carcinogenesis of ATC.
The gene encoding destrin behaved similarly in our experiments. Destrin is important for actin remodeling, endocytosis, polarized cell growth and cellular activation (Moriyama et al. 1990, Yahara et al. 1996). Chromosome 20p, where the destrin gene locates, is often amplified in ovarian cancer cell lines (Watanabe et al. 2001). Our results suggest that destrin might be a previously unsuspected participant in carcinogenesis. For its part, stathmin (Op18) is a member of a novel class of microtubule-destabilizing proteins that regulate the dynamics of microtubule polymerization and depolymerization (Mistry & Atweh 2002). Stathmin protein appears to have oncogenic potential, because it is widely expressed in various kinds of human cancers, including leukemia (Ghosh et al. 1993), prostate cancer (Friedrich et al. 1995) and breast cancer (Curmi et al. 2000), and because inhibition of stathmin can decrease the rate of proliferation of K562 erythroleukemic cells (Luo et al. 1994). In the clinical setting, some anti-cancer drug regimens are designed to inhibit microtubule assembly and arrest cells in mitosis, or to promote assembly of microtubules and stabilize tubulin polymers by preventing their depolymerization (Mistry & Atweh 2002). Over-expression of annexin II (15q21-22) reflects poor prognosis of colorectal and gastric cancers (Emoto et al. 2001a, b). Our results appear to corroborate an oncogenic role of annexin II in ATC.
On the other hand, 56 genes, including hypothetical protein and ESTs, were significantly down-regulated in ACLs on our microarray. Although one of them, thyroglobulin, serves as a tumor marker in differentiated thyroid cancers (Hoang-Vu et al. 1992), its expression was significantly decreased in ACL. Expression of TTF-1 (interacting peptide 20) also decreased in this study; the TTF-1 gene encodes a transcription factor that contributes to expression of thyroid-specific proteins like thyroglobulin (Fabbro et al. 1994). These results implied that a drastic abolition of normal thyroidal function had occurred in the tissue — giving rise to the parent ATCs, and suggested that thyroglobulin cannot be used as a marker for anaplastic thyroid tumors. PBP was also under-expressed. PBP, alternatively known as Raf-mediated activation inhibitor protein (RKIP), is expressed in prostate cancers but not in metastatic foci derived from those tumors. In other reports, over-expression of RKIP in C4-2B cells decreased cell invasion in vitro and inhibited lung metastasis (Fu et al. 2003). Thus PBP has shown a tumor-suppressive function in prostate cancer; our data suggest that PBP might be a tumor suppressor for human ATC as well. Of the 56 under-expressed genes listed in Table 2
, 22 genes are located in chromosomal regions where we previously detected LOH in > 20% of informative ATCs. In particular, CD34 (1q32), SHBG (17p13–p12) and autoantigen CALR (19p13.2) locate on the chromosomal position which showed frequent LOH in ATC (40%, 44% and 36% respectively) (Kitamura et al. 2000). These genes should be investigated as to potential function of tumor suppression in thyroid tissue.
Cell lines generally reflect the character of their tumors of origin but it is always possible for the nature of a cell line to change during immortalization. It is therefore necessary to evaluate expression profiles of primary tumors as well, but because ATC is so aggressive that there is little chance for surgical intervention which would provide fresh tissue for analysis. For the study reported here we did have access to ten precious ATC samples, and we were able to perform SQ-PCR with selected genes. Of 12 genes that had been significantly over-expressed in ACL, nine (destrin, HSPA8, stathmin, LDH-A, ATP5A1, PSMB6, B23, HDP-1 and LDH-B) were also over-expressed in primary ATCs. Of those, LDH-A and -B are isoenzymes, and LDH-A is reported to increase in tumors of all origins (Liu et al. 2003). Generally speaking, up-regulation of metabolic enzymes such as LDH-A or -B and ATP5A1 is likely to be the result of high metabolic activity of ACL, not a cause of carcinogenesis. B23 (alternatively NPM1) is a nucleolar phosphoprotein that is more abundant in tumors than in normal cells. Functionally, it relates to chromosomal translocation, and its over-expression has been confirmed in acute myeloid leukemia by microarray analysis (Jhanwar et al. 1984). HSPA8, PSMB6 and HDP-1, located at 12q23, 17p13 and 6p22.1 respectively, do not have any known functions, especially in cancer. On the other hand, up-regulation of NPD017, IDH3B and ANXA2 was found in ACLs only, suggesting that the activating changes might have been acquired in the process of immortalization.
Of the 15 genes that were selected from the list of under-expressed elements in ACL only three (thyroglobulin, PBP and FES) were under-expressed in primary tumors as well. Down-regulation of thyroglobulin might reflect the loss of normal thyroidal function in ATC. As the gene encoding PBP locates at 12q24-23, where LOH is frequent in ATCs (Kitamura et al. 2000), the combined evidence suggests that PBP might have tumor-suppressive function in thyroid tissue. The FES gene, at 15q26.1 (Lionberger & Smithgall 2000), exhibits strong expression in hematopoietic cells of the myeloid lineage and may regulate chronic myelogenous leukemia (Lionberger & Smithgall 2000). The meaning of the down-regulation of this apparent proto-oncogene in ACL and ATC is unclear; however, it might have a novel function in ATC carcinogenesis apart from the one it has in other situations.
The work reported here has yielded useful information for understanding the molecular mechanism(s) involved in ATC, and has revealed several novel genetic alterations that could be responsible for ATC carcinogenesis. The cause of up-regulation or down-regulation of these genes, and genetic or epigenetic change should be determined. Further study is required.
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Acknowledgements |
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References |
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TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
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Curmi PA, Nogues C, Lachkar S, Carelle N, Gonthier MP, Sobel A, Lidereau R & Bieche I 2000 Overexpression of stathmin in breast carcinomas points out highly proliferative tumours. British Journal of Cancer 82 142–150.[CrossRef][ISI][Medline]
Emoto K, Sawada H, Yamada Y, Fujimoto H, Takahama Y, Ueno M, Takayama T, Uchida H, Kamada K, Naito A et al. 2001a Annexin II overexpression is correlated with poor prognosis in human gastric carcinoma. Anticancer Research 21 1339–1345.[ISI][Medline]
Emoto K, Yamada Y, Sawada H, Fujimoto H, Ueno M, Takayama T, Kamada K, Naito A, Hirao S & Nakajima Y 2001b Annexin II overexpression correlates with stromal tenascin-C overexpression: a prognostic marker in colorectal carcinoma. Cancer 92 1419–1426.[CrossRef][ISI][Medline]
Fabbro D, Di Loreto C, Beltrami CA, Belfiore A, Di Lauro R & Damante G 1994 Expression of thyroid-specific transcription factors TTF-1 and PAX-8 in human thyroid neoplasms. Cancer Research 54 4744–4749.
Friedrich B, Gronberg H, Landstrom M, Gullberg M & Bergh A 1995 Differentiation-stage specific expression of oncoprotein 18 in human and rat prostatic adenocarcinoma. Prostate 27 102–109.[ISI][Medline]
Fu Z, Smith PC, Zhang L, Rubin MA, Dunn RL, Yao Z & Keller ET 2003 Effects of raf kinase inhibitor protein expression on suppression of prostate cancer metastasis. Journal of the National Cancer Institute 95 878–889.
Garcia-Rostan G, Tallini G, Herrero A, D’Aquila TG, Carcangiu ML & Rimm DL 1999 Frequent mutation and nuclear localization of beta-catenin in anaplastic thyroid carcinoma. Cancer Research 59 1811–1815.
Ghosh PK, Anderson J, Cohen N, Takeshita K, Atweh GF & Lebowitz P 1993 Expression of the leukemia-associated gene, p18, in normal and malignant tissues; inactivation of expression in a patient with cleaved B cell lymphoma/ leukemia. Oncogene 8 2869–2872.[ISI][Medline]
Hoang-Vu C, Dralle H, Scheumann G, Maenhaut C, Horn R, von zur Muhlen A & Brabant G 1992 Gene expression of differentiation- and dedifferentiation markers in normal and malignant human thyroid tissues. Experimental and Clinical Endocrinology 100 51–56.[ISI][Medline]
Jhanwar SC, Neel BG, Hayward WS & Chaganti RS 1984 Localization of the cellular oncogenes ABL, SIS, and FES on human germ-line chromosomes. Cytogenetics and Cell Genetics 38 73–75.[ISI][Medline]
Kitahara O, Katagiri T, Tsunoda T, Harima Y & Nakamura Y 2002 Classification of sensitivity or resistance of cervical cancers to ionizing radiation according to expression profiles of 62 genes selected by cDNA microarray analysis. Neoplasia 4 295–303.[CrossRef][ISI][Medline]
Kitamura Y, Shimizu K, Tanaka S, Ito K & Emi M 2000 Allelotyping of anaplastic thyroid carcinoma: frequent allelic losses on 1q, 9p, 11, 17, 19p, and 22q. Genes, Chromosomes and Cancer 27 244–251.[CrossRef][ISI][Medline]
Lionberger JM & Smithgall TE 2000 The c-Fes protein-tyrosine kinase suppresses cytokine-independent outgrowth of myeloid leukemia cells induced by Bcr-Abl. Cancer Research 60 1097–1103.
Liu ZJ, Peng WC, Yang X, Huang JF, Zhang XB, Zhang Y & Maekawa M 2003 Relative mRNA expression of the lactate dehydrogenase A and B subunits as determined by simultaneous amplification and single strand conformation polymorphism. Relation with subunit enzyme activity. Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences 793 405–412.
Luo XN, Mookerjee B, Ferrari A, Mistry S & Atweh GF 1994 Regulation of phosphoprotein p18 in leukemic cells. Cell cycle regulated phosphorylation by p34cdc2 kinase. Journal of Biological Chemistry 269 10312–10318.
Manos EJ & Jones DA 2001 Assessment of tumor necrosis factor receptor and Fas signaling pathways by transcriptional profiling. Cancer Research 61 433–438.
Mistry SJ & Atweh GF 2002 Role of stathmin in the regulation of the mitotic spindle: potential applications in cancer therapy. Mt Sinai Journal of Medicine 69 299–304.
Moriyama K, Nishida E, Yonezawa N, Sakai H, Matsumoto S, Iida K & Yahara I 1990 Destrin, a mammalian actin-depolymerizing protein, is closely related to cofilin. Cloning and expression of porcine brain destrin cDNA. Journal of Biological Chemistry 265 5768–5773.
Nakamura T, Yana I, Kobayashi T, Shin E, Karakawa K, Fujita S, Miya A, Mori T, Nishisho I & Takai S 1992 p53 gene mutations associated with anaplastic transformation of human thyroid carcinomas. Japanese Journal of Cancer Research 83 1293–1298.[CrossRef][ISI][Medline]
Ono K, Tanaka T, Tsunoda T, Kitahara O, Kihara C, Okamoto A, Ochiai K, Takagi T & Nakamura Y 2000 Identification by cDNA microarray of genes involved in ovarian carcinogenesis. Cancer Research 60 5007–5011.
Ozaki O, Ito K, Mimura T & Sugino K 1999 Anaplastic transformation of papillary thyroid carcinoma in recurrent disease in regional lymph nodes: a histologic and immunohistochemical study. Journal of Surgical Oncology 70 45–48.[CrossRef][ISI][Medline]
Passler C, Scheuba C, Prager G, Kaserer K, Flores JA, Vierhapper H & Niederle B 1999 Anaplastic (undifferentiated) thyroid carcinoma (ATC). A retrospective analysis. Langenbeck’s Archives of Surgery 384 284–293.
Saiz AD, Olvera M, Rezk S, Florentine BA, McCourty A & Brynes RK 2002 Immunohistochemical expression of cyclin D1, E2F-1, and Ki-67 in benign and malignant thyroid lesions. Journal of Pathology 198 157–162.
Sanchez-Carbayo M, Socci ND, Charytonowicz E, Lu M, Prystowsky M, Childs G & Cordon-Cardo C 2002 Molecular profiling of bladder cancer using cDNA microarrays: defining histogenesis and biological phenotypes. Cancer Research 62 6973–6980.
Sherman SI 2003. Thyroid carcinoma. Lancet 361 501–511.[CrossRef][ISI][Medline]
Speicher MR, Howe C, Crotty P, du Manoir S, Costa J & Ward DC 1995 Comparative genomic hybridization detects novel deletions and amplifications in head and neck squamous cell carcinomas. Cancer Research 55 1010–1013.
Voutilainen PE, Multanen M, Haapiainen RK, Leppaniemi AK & Sivula AH 1999 Anaplastic thyroid carcinoma survival. World Journal of Surgery 23 975–978; discussion 978–979.[CrossRef][ISI][Medline]
Watanabe T, Imoto I, Kosugi Y, Ishiwata I, Inoue S, Takayama M, Sato A & Inazawa J 2001 A novel amplification at 17q21–23 in ovarian cancer cell lines detected by comparative genomic hybridization. Gynecologic Oncology 81 172–177.[CrossRef][ISI][Medline]
Yahara I, Aizawa H, Moriyama K, Iida K, Yonezawa N, Nishida E, Hatanaka H & Inagaki F 1996 A role of cofilin/destrin in reorganization of actin cytoskeleton in response to stresses and cell stimuli. Cell Structure and Function 21 421–424.[ISI][Medline]
Yang YH, Dudoit S, Luu P, Lin DM, Peng V, Ngai J & Speed TP 2002 Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. Nucleic Acids Research 30 e15.
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