Molecular Cloning of the Canine Fragile Histidine Triad (FHIT) Gene and Fhit Protein Expression in Canine Peripheral Blood Mononuclear Cells

Hiraoka, Hiroko; Minami, Koji; Kaneko, Naoki; Miyama, Takako Shimokawa; Mizuno, Takuya; Okuda, Masaru

doi:10.1292/jvms.71.645

Cited by 3 publications

(8 citation statements)

References 22 publications

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“…We prepared 3 primer pairs-e1/eL, BF1/eL and TF5/TF3 (Table 1)-to detect the canine FHIT genes; in our previous study, we had confirmed that these 3 primer pairs amplify the canine FHIT gene transcripts [15]. These primer pairs were used to amplify exons 1 to 9 of the canine wild-type FHIT transcript, exons 3B to 9 of the alternative splicing form (type B), and exons 4 to 8 of the common ORF of these transcripts (Fig.…”

Section: Cellsmentioning

confidence: 89%

“…It has recently been shown that an alternative splicing form-the type-B FHIT transcript-is coexpressed with the wild-type FHIT transcript in the PBMCs, spleen and intestine of healthy dogs and that it may play a role in an alternative Fhit-protein-expressing system that is activated when expression of the wild-type transcript is reduced [15]. In this study, the type-B transcript could not be detected in any of the canine lymphoma cell lines.…”

Section: Discussionmentioning

confidence: 99%

“…Immunoblot analysis: Immunoblot analysis was performed by using a rabbit antihuman Fhit polyclonal antibody (clone ZR44; Invitrogen) that had been confirmed to detect the canine Fhit protein in a previous study [15]. Each sample was lysed in NP-40 lysis buffer [1% NP-40, 50 mM Tris (pH 8.0), 150 mM NaCl, 1% SDS, 2 g/ml leupeptin and 2 g/ml aprotinin] and was sonicated to degrade the genomic DNA.…”

Section: Cellsmentioning

confidence: 99%

“…Expression of the Fhit protein in canine lymphoma cell lines: In our previous study, a rabbit antihuman Fhit polyclonal antibody (clone ZR44, Invitrogen) was used to detect the canine Fhit protein (approximately 17 kDa) in the PBMCs of healthy dogs [15]. We performed immunoblot analysis by using this antibody in order to evaluate expression of the Fhit protein in canine lymphoma cell lines.…”

Section: Expression Of Fhit Transcripts In Canine Lymphoma Cell Linesmentioning

confidence: 99%

“…Recently, we demonstrated the canine FHIT gene structure and its protein expression in peripheral blood mononuclear cells (PBMCs) and other tissues, including tissues of the lungs, intestine, liver, spleen and thymus, in healthy dogs [15]. The canine FHIT gene is located in an approximately 1.4-megabase pair (Mbp) region of canine chromosome 20; the wild-type transcript is composed of 9 small exons (30-306 bp) that are separated by extremely large introns (2,333-494,789 bp), and the ORF corresponds to exons 4 to 8.…”

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confidence: 99%

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Aberrations of the FHIT Gene and Fhit Protein in Canine Lymphoma Cell Lines

Hiraoka

Minami

Kaneko

et al. 2009

The Journal of Veterinary Medical Science

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ABSTRACT. The fragile histidine triad (FHIT) gene is a tumor-associated gene, and aberrant FHIT gene and protein expression have been described in many types of human tumors. Furthermore, it has been reported that FHIT gene inactivation is induced by hypermethylation of 5' CpG islands in the gene or by genomic deletion around the open reading frame (ORF). In this study, we explored the aberrations in the canine FHIT gene and Fhit protein expression and assessed the methylation status and genomic deletions by using 5 canine lymphoma cell lines. We found that the decrease in the expression of the Fhit protein in canine lymphoma cell lines was similar to that in human tumors. The expression of the wild-type FHIT transcript was reduced in all 5 cell lines. However, we could not confirm the involvement of aberrant methylation events in the 5' CpG islands of the canine FHIT gene. We were able to identify homozygous or heterozygous deletions in the canine FHIT genes in all 5 cell lines. Moreover, a widespread genomic deletion of the FHIT gene, which included the ORF region, was detected in 1 cell line. In the present study, we detected aberrations in the FHIT gene and Fhit protein expression in all 5 canine lymphoma cell lines, and this phenomenon might be an important factor in promoting canine lymphoma.

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Section: Cellsmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Section: Cellsmentioning

confidence: 99%

Section: Expression Of Fhit Transcripts In Canine Lymphoma Cell Linesmentioning

confidence: 99%

mentioning

confidence: 99%

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Aberrations of the FHIT Gene and Fhit Protein in Canine Lymphoma Cell Lines

Hiraoka

Minami

Kaneko

et al. 2009

The Journal of Veterinary Medical Science

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Fragile histidine triad protein: structure, function, and its association with tumorogenesis

Hassan

Naiyer

Ahmad

2009

J Cancer Res Clin Oncol

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In silico study of fragile histidine triad interaction domains with MDM2 and p53

2014

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Background:Fragile histidine triad (FHIT) is considered as a member of the histidine triad (HIT) nucleotide-binding protein superfamily regarded as a putative tumor suppressor executing crucial role in inhibiting p53 degradation by MDM2. Accumulating evidences indicate FHIT interaction with p53 or MDM2; however, there is no certain study deciphering functional domains of FHIT involving in the interaction with MDM2 and/or p53. In this regard, such evident interaction can spring in mind determining important domains of FHIT binding to MDM2 with regard to p53.Materials and Methods:Since there were not any previous studies appraising complete three-dimensional structures of target molecules, molecular modeling was carried out to construct three-dimensional models of full FHIT, MDM2, P53 and also FHIT segments. Truncated structures of FHIT were created to reveal critical regions engaging in FHIT interaction.Results:Given the shape and shape/electrostatic total energy, FHIT structures (β1-5), (β3-7, α1), and (β5-7, α1) appeared to be better candidates than other structures in interaction with full MDM2. Furthermore, FHIT structures (β6-7), (β6-7, α1), (β4-7, α1) were considered to be better than other structures in interaction with p53. FHIT truncates that interact with MDM2 presented lower energy levels than FHIT truncates interacting with p53.Conclusion:These findings are beneficial to understand the mechanism of the FHIT-MDM2-p53 complex activation for designing inhibitory compounds.

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Molecular Cloning of the Canine Fragile Histidine Triad (FHIT) Gene and Fhit Protein Expression in Canine Peripheral Blood Mononuclear Cells

Cited by 3 publications

References 22 publications

Aberrations of the FHIT Gene and Fhit Protein in Canine Lymphoma Cell Lines

Aberrations of the FHIT Gene and Fhit Protein in Canine Lymphoma Cell Lines

Fragile histidine triad protein: structure, function, and its association with tumorogenesis

In silico study of fragile histidine triad interaction domains with MDM2 and p53

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