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SLUG silencing increases radiosensitivity of melanoma cells in vitro

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Abstract

Background

Melanoma radioresistance has been attributed to the presence of tumor cells with highly efficient DNA damage repair mechanisms. We examined the expression of genes involved in DNA damage repair and DNA damage sensing, and assessed their modulation by SLUG silencing, which is potentially capable of increasing radiosensitivity.

Methods

Two melanoma cell lines (M14 and M79) were used to evaluate in vitro radiation-induced cytotoxicity before and after SLUG silencing. mRNA expression levels of BRCA1, ERCC1, DNA-PK, PARP, MGMT, ATM and TGM2 were determined by real-time RT-PCR, and protein expression levels of SLUG, caspase 3, p21, PUMA and pMAPK by Western blotting.

Results

The cytotoxic effect of radiation was high in M14 and low in M79 cells. SLUG silencing increased the interference of radiation on cell cycle distribution and cell killing by 60 % and 80 % in M79 cells after a 2.4 Gy and 5 Gy radiation dose, respectively. It also led to a significant inhibition of expression of genes involved in DNA damage repair and DNA damage sensing in all cell lines maintained after radiation. An almost total inhibition was observed for TGM2, which is expressed at a high basal level in the most radioresistant cell line (M79). Protein expression of PUMA was induced by radiation and was enhanced after SLUG silencing.

Conclusions

Our results reveal a pivotal role of SLUG in regulating a cellular network involved in the response to DNA damage, and highlight the importance of TGM2 in radiosensitivity modulation. SLUG silencing appears to increase radiation sensitivity of the melanoma cells tested.

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Acknowledgement

The authors thank Ursula Elbling for editing the manuscript.

Conflicts of interest

There are no conflicts of interest to declare.

Funding

This work was supported by a grant from the Italian Ministry of Health (RO Strategici 11/07) “p53 family interaction network as a target of antitumoral peptide therapy”. The funding body did not have a role in the study design, in the collection, analysis and interpretation of data; in the writing of the manuscript; or in the decision to submit the manuscript for publication.

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Authors and Affiliations

  1. Biosciences Laboratory, IRCCS Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST), Via P. Maroncelli 40, Meldola, 47014, Italy

    Chiara Arienti, Anna Tesei, Silvia Carloni, Paola Ulivi, Rosella Silvestrini & Wainer Zoli

  2. Radiotherapy Unit, IRCCS Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST), Meldola (FC), Italy

    Antonino Romeo, Giulia Ghigi & Elisabetta Parisi

  3. Health Physics Unit, IRCCS Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST), Meldola (FC), Italy

    Enrico Menghi & Anna Sarnelli

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  1. Chiara Arienti
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Correspondence to Wainer Zoli.

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Arienti, C., Tesei, A., Carloni, S. et al. SLUG silencing increases radiosensitivity of melanoma cells in vitro. Cell Oncol. 36, 131–139 (2013). https://doi.org/10.1007/s13402-012-0120-6

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  • DOI: https://doi.org/10.1007/s13402-012-0120-6

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