Expression profiles of 39 HOX genes in normal human adult organs and anaplastic thyroid cancer cell lines by quantitative real-time RT-PCR system

Takahashi, Yōko; Hamada, Jun Ichi; Murakawa, Katsuhiko; Takada, Minoru; Tada, Mitsuhiro; Nogami, Ikuko; Hayashi, Nobuyasu; Nakamori, Shoji; Monden, Morito; Miyamoto, Masaki; Katoh, Hiroyuki; Moriuchi, Tetsuya

doi:10.1016/j.yexcr.2003.09.024

Cited by 145 publications

(133 citation statements)

References 35 publications

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“…Real-time PCR amplification was performed in 20 L reaction mixture containing 2 L cDNA sample, 10 L QuantiTect SYBR Green PCR Master Mix (Qiagen, Valencia, CA) and with specific primer sets that had been described in our previous report. 9 PCR was carried out by starting with a 15-min hot start at 95°C followed by a denaturation step at 94°C for 15 sec, an annealing step at 60°C for 30 sec and an extension step at 72°C for 1 min for 40 cycles. Dissociation curve analysis (95°C for 15 sec, 60°C for 15 sec and 95°C for 15 sec) was performed at the end of 40 cycles to verify the PCR product identity.…”

Section: Quantitative Real-time Rt-pcrmentioning

confidence: 99%

“…Quantification was done by using the standard curve method according to the method described in our previous report. 9 Finally, we represented relative gene expression levels as the ratio of the target HOX gene to the internal reference gene (␤-actin) expression based on the initial copy number calibrated along the standard curve.…”

Section: Quantitative Real-time Rt-pcrmentioning

confidence: 99%

“…Y-axis represents relative expression as the ratio of the target HOX gene to the internal reference gene (␤-actin) expression based on the initial copy number calibrated along the standard curve as described in text and in our previous report. 9 …”

Section: Difference In Hox Gene Expression Among Melanoma With Differmentioning

confidence: 99%

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Altered expressions of HOX genes in human cutaneous malignant melanoma

Maeda

Hamada

Takahashi

et al. 2004

Intl Journal of Cancer

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HOX genes act as master genes to control morphogenesis. In human, HOX genes form 4 clusters composing 9 to 11 HOX genes (39 genes in total) on different chromosomes. We hypothesized that aberrant expression of HOX genes was associated with development and subsequent progression of melanoma and that the 39 HOX gene expression pattern determined the sites where melanoma grew. The expression levels of 39 HOX genes in 15 human cutaneous melanoma specimens and 7 nevus pigmentosus specimens were quantified by a comprehensive analysis system based on the real-time RT-PCR method. We found that the expression levels of HOXA11, A13, B9, D12 and D13 in melanoma were higher than those in nevus pigmentosus and that the expression levels of HOXA11, B2 and C13 were significantly different between pT4 melanoma and pT1 to pT3 melanoma. It was most notable that the expression levels of HOXA1, A2, C4 and B13 in melanoma with distant metastasis were higher than those in melanoma without it. On the other hand, we found no relationship between HOX genes expression patterns and the growing sites of melanoma. These results indicated that the misexpressions of some specific HOX genes were implicated in melanoma genesis and metastasis but had no linkage with melanoma sites. © 2004 Wiley-Liss, Inc. Key words: melanoma; HOX; metastasis; nevus pigmentosusHox genes are one of the master regulators of morphogenesis and cell differentiation during embryogenesis of animals. 1 The genes contain a 180 bp DNA sequence (homeobox), which encodes a highly conserved 60 amino acid homeodomain. The HOX proteins function as transcription factors through their homeodomain, which is responsible for recognition and binding of sequence-specific DNA motifs. 2,3 In human genome, 39 HOX genes are clustered in a similar arrangement of 13 paralog groups in 4 different chromosomal regions, HOXA, B, C and D. 4 During embryonic morphogenesis, the HOX genes determine positional identity along the anterior-posterior and secondary axes in animals. Within each cluster, HOX genes located at the 3Ј extremity are activated first and in anterior embryonic domains, whereas 5Ј-located genes are transcribed subsequently and in more caudal areas, which is the property called colinearity rule. 5 HOX genes are also expressed in some normal adult organs in characteristic patterns, suggesting their possible role in the maintenance of tissuespecific architecture besides their roles in embryogenesis. 6 -9 Comparative analysis of HOX gene expression between cancerous tissues and normal tissues uncovered the dysregulated expression(s) of a particular HOX gene(s) in some kinds of carcinomas. For example, HOXB5 and B9 are expressed in normal kidney but not in renal cancer, whereas the expression of HOXC11 is observed in human renal cancer but not in normal kidney. 6 In human prostate cancer, overexpression of HOXC8 correlates with loss of differentiated phenotype. 10 Increased expressions of HOXC4, C5, C6 and C11 are likely to be involved in the development of human bladder transitiona...

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Section: Quantitative Real-time Rt-pcrmentioning

confidence: 99%

Section: Quantitative Real-time Rt-pcrmentioning

confidence: 99%

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Altered expressions of HOX genes in human cutaneous malignant melanoma

Maeda

Hamada

Takahashi

et al. 2004

Intl Journal of Cancer

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“…While their role in animal development is well established, their role in evolution is less well understood, see e.g. [12,69]. A particularly intriguing problem is the role of Hox cluster duplications in vertebrate evolution.…”

Section: Introductionmentioning

confidence: 99%

The duplication of the Hox gene clusters in teleost fishes*1

Prohaska

Stadler

2004

Theory in Biosciences

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Higher teleost fishes, including zebrafish and fugu, have duplicated their Hox genes relative to the gene inventory of other gnathostome lineages. The most widely accepted theory contends that the duplicate Hox clusters orginated synchronously during a single genome duplication event in the early history of ray-finned fishes. In this contribution we collect and re-evaluate all publicly available sequence information. In particular, we show that the short Hox gene fragments from published PCR surveys of the killifish Fundulus heteroclitus, the medaka Oryzias latipes and the goldfish Carassius auratus can used to determine with little ambiguity not only their paralog group but also their membership in a particular cluster. Together with a survey of the genomic sequence data from the pufferfish Tetraodon nigroviridis we show that at least percomorpha, and possibly all eutelosts, share a system of seven orthologous Hox gene clusters, while at least the HoxC and HoxD clusters in ostariophysian (zebrafish) lineage might have arisen independently. There is little doubt about the orthology of the two teleost duplicates of the HoxA and HoxB clusters. A careful analysis of both the coding sequence of Hox genes and of conserved noncoding sequences provides additional support for the "duplication early" hypothesis that the Hox clusters in teleosts are derived by subsequent gene loss from an eight-cluster situation, although the data remain ambiguous in particular for the HoxC clusters. Assuming the "duplication early" hypothesis we use the new evidence on the Hox gene complements to determine the phylogenetic positions of gene-loss events in the wake of the cluster duplication. Surprisingly, we find that the resolution of redundancy seems to be a slow process that ist still-ongoing. A few suggestion on which additional sequence data would be most informative for resolving the history of the teleostean Hox genes are discussed.

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“…For exclusion of contaminated genomic DNA, 50 μg of total RNA was incubated for 30 min at 37˚C in 50 μl of reaction mixture containing 40 mM TrisHCl (pH 7.2), 10 mM NaCl, 6 mM MgCl 2 , 2 mM dithiothreitol, 0.04 U/μl PQ1 DNase (Promega, Madison, WI), and 0.4 U/μl RNase inhibitor. Reverse transcription reaction for real-time PCR was performed by the method described in our previous study (22). Table I.…”

Section: Methodsmentioning

confidence: 99%

Dysregulated expression of HOX and ParaHOX genes in human esophageal squamous cell carcinoma

et al. 2007

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Abstract. Homeobox genes function as master regulators in embryonic morphogenesis. We hypothesized that homeobox genes are essential to maintain tissue-or organ-specificity even in adult body and that the dysregulated expression of homeobox genes results in tumor development and progression. To better understand the roles of homeobox genes in development and progression of esophageal cancer, we analyzed the expression patterns of 39 HOX genes and 4 ParaHOX (CDX1, CDX2, CDX4 and PDX1) genes in esophageal squamous cell carcinoma (ESCC) and normal esophageal mucosa tissues. A total of 48 primary ESCC tissues and 7 normal esophageal mucosa tissues were resected from patients who underwent radical surgery without any preoperative chemotherapy or radiotherapy. The expression of HOX and ParaHOX genes were analyzed by a quantitative real-time RT-PCR method and immunohistochemistry. The expression levels of 24 HOX genes, CDX1, CDX2 and PDX1 were significantly higher in ESCC compared to normal mucosa (p<0.01, Mann-Whitney U test). The Immunohistochemical study revealed that HOXA5 and D9 proteins were more cytoplasmic in ESCC than normal mucosa cells. Our data indicate that the disordered expression of HOX and ParaHOX genes are involved in the development of ESCC or its malignancy.

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Expression profiles of 39 HOX genes in normal human adult organs and anaplastic thyroid cancer cell lines by quantitative real-time RT-PCR system

Cited by 145 publications

References 35 publications

Altered expressions of HOX genes in human cutaneous malignant melanoma

Altered expressions of HOX genes in human cutaneous malignant melanoma

The duplication of the Hox gene clusters in teleost fishes*1

Dysregulated expression of HOX and ParaHOX genes in human esophageal squamous cell carcinoma

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