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T[20] repeat in the 3′-untranslated region of the MT1X gene: a marker with high sensitivity and specificity to detect microsatellite instability in colorectal cancer

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International Journal of Colorectal Disease Aims and scope Submit manuscript

Abstract

Purpose

Stratifying patients defective in mismatch repair (dMMR) with high microsatellite instability (MSI-H) in colorectal cancer (CRC) is of increasing relevance and may provide a more tailored approach to CRC adjuvant therapy. Here, we describe the discovery of a new MSI marker for colorectal cancer located in the 3′-untranslated region (3′UTR, T20 mononucleotide repeat) of the metallothionein 1X gene (MT1XT20).

Methods

We studied 340 consecutive CRCs using three multiplexed polymerase chain reactions amplifying BAT25, BAT26, TGFBR2, MybT22, BAT40, MT1XT20, NR21, NR24, CAT25, D2S123, D5S346, D17S250, D18S58, CSF1PO, D7S820, and D18S51. Fragments length was evaluated by automated capillary electrophoresis.

Results

Based on the NCI/ICG-HNPCC criteria for MSI classification, 40 CRCs were found to be MSI-high (11.8%), 46 (13.5%) CRCs were MSI-low, and 254 CRCs (74.7%) were stable (MSS). MT1XT20 showed very high sensitivity (97.3%) comparable to BAT26 (97.5%) and CAT25 (97.1%) and the best specificity (100%) as well as MybT22 and CAT25. Indeed, MT1XT20 instability was detected in 36 out of 37 cases (97.3%) of MSI-high colorectal cancers, whereas no MT1XT20 alterations were observed in 254 MSS or in 46 MSI-low cases. On the contrary, BAT40 was found to be unstable in 8/46 MSI-low cases, BAT25 in 6/46, BAT26 4/46, NR21 1/46, and NR24 in 1/45.

Conclusions

Our results suggest that MT1XT20 represents a sensitive and specific marker for MSI testing and could be included in a complete set of MSI markers for the confident identification of familial or sporadic dMMR patients in CRCs.

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Abbreviations

dMMR:

Defective mismatch repair

MSI-H:

High microsatellite instability

CRC:

Colorectal cancer

MT1XT20:

3′UTR T20 mononucleotide repeat of the metallothionein 1X gene

MSI-L:

Low microsatellite instability

MSS:

Microsatellite stability

NCI/ICG-HNPCC:

National Cancer Institute/International Collaborative Group-Hereditary Non-Polyposis Colorectal Cancer

AJCC/UICC:

American Joint Cancer Committee/Union Internationale Contre le Cancer

FFPE:

In formalin fixed and paraffin embedded

H&E:

Haematoxylin and eosin

LOH:

Loss of heterozygosity

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Acknowledgements and funding

This study was supported in part by a regional grant (1897, 2008) given by Friuli Venezia Giulia to Alphagenics Diaco Biotechnologies S.r.l., Area Science Park, Basovizza, Trieste Italy. DdB is founded by CIRC (Centro Interdipartimentale di Ricerca sul Cancro, University of Bologna).

Competing interests

Luca Morandi declares that he has applied for a patent relating to the content of this manuscript. Adriana Monzoni is affiliated with Alphagenics Diaco Biotechnologies S.r.l. and is full-time employed. The other authors indicated no potential conflicts of interest.

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Authors and Affiliations

  1. Department of Haematology and Oncological Sciences L. and A. Seragnoli, Section of Anatomic Pathology at Bellaria Hospital, University of Bologna, via Altura 3, 40139, Bologna, Italy

    Luca Morandi, Dario de Biase, Michela Visani, Annalisa Tosi, Paola Baccarini & Giovanni Tallini

  2. Department of Experimental Pathology, University of Bologna, via San Giacomo 14, 40126, Bologna, Italy

    Dario de Biase, Michela Visani & Annalisa Pession

  3. Alphagenics Diaco Biotechnologies S.r.l., Area Science Park SS14, Basovizza, Trieste, Italy

    Adriana Monzoni

  4. General Surgery, Bellaria Hospital A.U.S.L. Bologna, via Altura 3, 40139, Bologna, Italy

    Mauro Brulatti

  5. Medical Genetics Unit, Polyclinic S. Orsola-Malpighi, University of Bologna, Bologna, Italy

    Daniela Turchetti

  6. Sezione di Anatomia Patologica “M. Malpighi”, Dipartimento di Ematologia e Scienze Oncologiche “L. e A. Seragnoli”, Università di Bologna, Ospedale Bellaria, via Altura 3, 40139, Bologna, Italy

    Luca Morandi

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  1. Luca Morandi
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Correspondence to Luca Morandi.

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ESM

MSI status in detail for our series: sheet 1 (MSI-high) is a list of all 40 MSI-H specimens; sheet 2 (MSI-low) is a list of all 46 MSI-L specimens. MSI microsatellite instability, MSS microsatellite stability, LOH loss of heterozygosity, AI allelic imbalance, NA not available (XLS 59 kb)

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Morandi, L., de Biase, D., Visani, M. et al. T[20] repeat in the 3′-untranslated region of the MT1X gene: a marker with high sensitivity and specificity to detect microsatellite instability in colorectal cancer. Int J Colorectal Dis 27, 647–656 (2012). https://doi.org/10.1007/s00384-011-1365-7

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