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Thymidylate synthetase allelic imbalance in clear cell renal carcinoma

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Abstract

Purpose

To investigate the allelic status of the thymidylate synthetase (TYMS) gene, located at chromosome band 18p11.32, in renal cell carcinoma (RCC). TYMS is a key target of the 5-fluorouracil (5-FU)-based class of drugs, frequently considered in combination therapies in advanced RCC. TYMS variants, such as the TYMS polymorphic 5′-untranslated region variable number tandem repeat sequence (VNTR), are under investigation to guide 5-FU treatment. Yet, no information is available with regard to changes in TYMS allele frequencies in RCC malignances.

Methods

Blood and matched tumor samples were collected from 41 histological proven clear cell RCC affected patients (30 males, 11 females.). TYMS VNTR genotype was first determined in blood to identify heterozygotes employing PCR techniques. To evaluate for allelic imbalance, fragment analysis was performed both in blood and matched tumor DNA of the heterozygote patients. Microsatellite analysis, employing the markers D18S59 and D18S476 mapping, respectively, at the TYMS locus (18p11.32) and 1.5 Mb downstream of the TYMS gene sequence (18p11.31), was performed to confirm TYMS allelic imbalance in tumors.

Results

Germ-line TYMS VNTR distribution was: 2R/2R (19.5%), TYMS 2R/3R (36.6%) and TYMS 3R/3R (43.9%). Allelic imbalance for the TYMS tandem repeat region was detected in 26.6% of the heterozygote patients. Microsatellite analysis confirmed the allelic imbalance detected by TYMS VNTR analysis and revealed that the overall frequence of allelic imbalance of chromosome band 18p11.32 was 35%, while the overall allelic imbalance of chromosome band 18p11.31 was 28%.

Conclusions

By focusing on the TYMS polymorphic variants in renal cancer, we here provide evidence, to our knowledge, for the first time showing loss of 18p11.32 and 18p11.31 in renal cell carcinomas. As allelic imbalances involving TYMS locus may be an important variable affecting 5-FU responsiveness, this study may contribute to explain different responses of advanced RCC in combined chemotherapeutic regimens incorporating fluoropyridines.

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Acknowledgments

The present work is a part of the study program supported by AOI (Associazione Oncologica Italiana no profit, Padova), IAES (International Academy of Environmental Sciences no profit, Venezia), Antonio R. Cananzi no profit Association (Padova) and Italian research program Biotech I and Biotech II (regione Veneto).

Conflict of interest statement

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Authors and Affiliations

  1. Research and Innovation (R&I) S.p.A., Via Svizzera 16, 35127, Padua, Italy

    Davide Colavito, Anna Stecca, Maurizio Dalle Carbonare & Alberta Leon

  2. 1ST UOC Medical Oncology, National Cancer Institute IOV-IRCCS, Via Gattamelata 64, 35128, Padua, Italy

    Giuseppe Cartei, Fable Zustovich, Miriam Farina & Eva Colombrino

  3. UOC Urology, S. Antonio Hospital, Via Facciolati 71, 35126, Padua, Italy

    Massimo Dal Bianco

  4. International Academy Environmental Sciences (IAES), Campo della Chiesa 3, 30132, Venice, Italy

    Giuseppe Cartei

  5. Department of Pharmacology and Anesthesiology, University of Padua, Largo Meneghetti 2, 35131, Padua, Italy

    Eugenio Ragazzi

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  1. Davide Colavito
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  2. Giuseppe Cartei
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  3. Massimo Dal Bianco
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  4. Anna Stecca
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  10. Alberta Leon
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Correspondence to Davide Colavito.

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Colavito, D., Cartei, G., Dal Bianco, M. et al. Thymidylate synthetase allelic imbalance in clear cell renal carcinoma. Cancer Chemother Pharmacol 64, 1195–1200 (2009). https://doi.org/10.1007/s00280-009-0986-9

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  • DOI: https://doi.org/10.1007/s00280-009-0986-9

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