Molecular cloning of the canine telomerase reverse transcriptase gene and its expression in neoplastic and non-neoplastic cells

Mitsuhiro YazawaDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Masaru OkudaLaboratory of Veterinary Internal Medicine, Faculty of Agriculture, Yamaguchi University, Yamaguchi, 753-8515, Japan.

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Noriko KanayaDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Sung-Hyeok HongDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Tomoko TakahashiDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Emi OhashiDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Takayuki NakagawaDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Ryohei NishimuraDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Nobuo SasakiDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Kenichi MasudaDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Koichi OhnoDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Hajime TsujimotoDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan.

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Abstract

Objective—To perform molecular cloning of the canine telomerase reverse transcriptase (TERT) gene and determine its expression in neoplastic and nonneoplastic cells.

Sample Population—9 canine tumor cell lines derived from various neoplasms, 16 primary canine tumors, and tissues from 15 normal canine organs.

Procedure—Tumor cell lines were derived from canine tumors that included osteosarcoma, mammary gland adenocarcinoma, melanoma, acute lymphoblastic leukemia, lymphoma, and mastocytoma and a canine primary fibroblast culture. Canine TERT complementary DNA (cDNA) was amplified by use of polymerase chain reaction (PCR) and sequenced. Expression of TERT mRNA was examined by reverse transcription (RT)-PCR assay. Telomerase activity was measured by use of the telomeric repeat amplification protocol assay.

Results—The canine TERT cDNA clone was 237 base pairs in length and contained a central region encoding the reverse transcriptase motif 2. Expression of TERT mRNA was detected in canine tumor cell lines that had telomerase activity but not in telomerasenegative canine primary fibroblasts. The TERT mRNA was detected in 13 of 16 canine tumor tissues and several normal tissues such as liver, ovary, lymph node, and thymus. A significant correlation between TERT expression level and telomerase activity was noted.

Conclusions and Clinical Relevance—Expression of TERT mRNA was closely associated with telomerase activity in neoplastic cells as well as some non-neoplastic cells from dogs. In addition to telomerase activity, expression of TERT mRNA can be used as a marker of tumor cells. (Am J Vet Res 2003;64:1395–1400)

Abstract

Objective—To perform molecular cloning of the canine telomerase reverse transcriptase (TERT) gene and determine its expression in neoplastic and nonneoplastic cells.

Sample Population—9 canine tumor cell lines derived from various neoplasms, 16 primary canine tumors, and tissues from 15 normal canine organs.

Procedure—Tumor cell lines were derived from canine tumors that included osteosarcoma, mammary gland adenocarcinoma, melanoma, acute lymphoblastic leukemia, lymphoma, and mastocytoma and a canine primary fibroblast culture. Canine TERT complementary DNA (cDNA) was amplified by use of polymerase chain reaction (PCR) and sequenced. Expression of TERT mRNA was examined by reverse transcription (RT)-PCR assay. Telomerase activity was measured by use of the telomeric repeat amplification protocol assay.

Results—The canine TERT cDNA clone was 237 base pairs in length and contained a central region encoding the reverse transcriptase motif 2. Expression of TERT mRNA was detected in canine tumor cell lines that had telomerase activity but not in telomerasenegative canine primary fibroblasts. The TERT mRNA was detected in 13 of 16 canine tumor tissues and several normal tissues such as liver, ovary, lymph node, and thymus. A significant correlation between TERT expression level and telomerase activity was noted.

Conclusions and Clinical Relevance—Expression of TERT mRNA was closely associated with telomerase activity in neoplastic cells as well as some non-neoplastic cells from dogs. In addition to telomerase activity, expression of TERT mRNA can be used as a marker of tumor cells. (Am J Vet Res 2003;64:1395–1400)