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MSH3 expression does not influence the sensitivity of colon cancer HCT116 cell line to oxaliplatin and poly(ADP-ribose) polymerase (PARP) inhibitor as monotherapy or in combination

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Abstract

Purpose

Defective expression of the mismatch repair protein MSH3 is frequently detected in colon cancer, and down-regulation of its expression was found to decrease sensitivity to platinum compounds or poly(ADP-ribose) polymerase inhibitors (PARPi) monotherapy. We have investigated whether MSH3 transfection in MSH3-deficient colon cancer cells confers resistance to oxaliplatin or PARPi and whether their combination restores chemosensitivity.

Methods

MSH3-deficient/MLH1-proficient colon cancer HCT116MLH1 cells were transfected with the MSH3 cDNA cloned into the pcDNA3.1(−) vector. MSH3/MLH1-deficient HCT116, carrying MLH1 and MSH3 mutations on chromosome 3 and 5, respectively, and HCT116 in which wild-type MLH1 (HCT116+3), MSH3 (HCT116+5) or both genes (HCT116+3+5) were introduced by chromosome transfer were also tested. Sensitivity to oxaliplatin and to PARPi was evaluated by analysis of clonogenic survival, cell proliferation, apoptosis and cell cycle.

Results

MSH3 transfection in HCT116 cells did not confer resistance to oxaliplatin or PARPi monotherapy. MSH3-proficient HCT116+5 or HCT116+3+5 cells, which were more resistant to oxaliplatin and PARPi in comparison with their MSH3-deficient counterparts, expressed higher levels of the nucleotide excision repair ERCC1 and XPF proteins, involved in the resistance to platinum compounds, and lower PARP-1 levels. In all cases, PARPi increased sensitivity to oxaliplatin.

Conclusions

Restoring of MSH3 expression by cDNA transfection, rather than by chromosome transfer, did not affect colon cancer sensitivity to oxaliplatin or PARPi monotherapy; PARP-1 levels seemed to be more crucial for the outcome of PARPi monotherapy.

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Acknowledgments

This work was supported by a grant from the Italian Ministry of Education and Research, “Programmi di ricerca scientifica di Rilevante Interesse Nazionale” (PRIN) project to LT. A.M. is recipient of a fellowship from “Regione Lazio-Filas”. The funders had no role in study design, data collection and analysis, or preparation of the manuscript.

Conflict of interest

The authors declare that they have no competing interests.

Author information

Authors and Affiliations

  1. Department of System Medicine, University of Rome “Tor Vergata”, Via Montpellier 1, 00133, Rome, Italy

    Lucio Tentori, Alessia Muzi, Annalisa Susanna Dorio & Grazia Graziani

  2. Department of Biomedicine, University of Rome “Tor Vergata”, Via Montpellier 1, 00133, Rome, Italy

    Susanna Dolci & Federica Campolo

  3. Department of Science and Technology of Education, University of Rome “Tor Vergata”, Via Montpellier 1, 00133, Rome, Italy

    Patrizia Vernole

  4. Laboratory of Molecular Oncology, “Istituto Dermopatico Dell’Immacolata (IDI), Istituto di Recovero e Cura a Carattere Scientifico (IRCCS)”, Via Monti di Creta 104, 00167, Rome, Italy

    Pedro Miguel Lacal

  5. INSERM, UMR_S 938, Saint-Antoine Research Centre, 75012, Paris, France

    Françoise Praz

  6. UPMC Univ Paris 06, UMR_S 938, Saint-Antoine Research Centre, 75012, Paris, France

    Françoise Praz

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  1. Lucio Tentori
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Correspondence to Grazia Graziani.

Electronic supplementary material

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280_2013_2175_MOESM1_ESM.pptx

Fig. S1 Analysis of apoptosis and cell cycle in MLH1-deficient and MLH1-proficient HCT116 cells treated with oxaliplatin. Cells were exposed to 1.2 and 2.5 μM oxaliplatin (Oxa) and analysed by flow cytometry at 72 h after treatment. The percentages of apoptotic cells are indicated. The percentages of MLH1-deficient cells in the different phases of cell cycle are as follows: Ctr, G1 41%, S 42%, G2/M 17%; Oxa 1.2 μM, G1 56%, S 17%, G2/M 27%; Oxa 2.5 μM, G1 52%, S 13%, G2/M 35%. The percentages of MLH1-proficient cells in the different phases of cell cycle are as follows: untreated control (Ctr), G1 54%, S 29%, G2/M 17%; Oxa 1.2 μM, G1 56%, S 7%, G2/M 37%; Oxa 2.5 μM, G1 52%, S 6%, G2/M 42% (PPTX 307 kb)

280_2013_2175_MOESM2_ESM.pptx

Fig. S2 Cell cycle analysis of HCT116+3 and HCT116+3+5 cells treated with PARPi. HCT116+3 and HC116+3+5 cells were exposed to GPI 15427 (5 μM) and analysed by flow cytometry at 72 h after treatment. The percentages of cells in the different phases of cell cycle are as follows: HCT116+3, Ctr, G1 52%, S 32%, G2/M 16%; HCT116+3, PARPi, G1 53%, S 29%, G2/M 18%; HCT116+3+5, Ctr, G1 54%, S 24%, G2/M 22%; HCT116+3+5, PARPi, G1 52%, S 22%, G2/M 26% (PPTX 121 kb)

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Tentori, L., Muzi, A., Dorio, A.S. et al. MSH3 expression does not influence the sensitivity of colon cancer HCT116 cell line to oxaliplatin and poly(ADP-ribose) polymerase (PARP) inhibitor as monotherapy or in combination. Cancer Chemother Pharmacol 72, 117–125 (2013). https://doi.org/10.1007/s00280-013-2175-0

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