Chemogenomic study of gemcitabine using Saccharomyces cerevisiae as model cell—molecular insights about chemoresistance

Abstract

Gemcitabine (GEM) is the drug used as first line to treat pancreatic cancer, one of the most devastating human tumors. This peculiar type of tumor develops resistance to several drugs, including GEM, due to its desmoplastic reaction and other features. The GEM chemoresistance has been investigated at molecular level aiming to find a pathway whose inhibition or activation should overcome it. Through next-generation sequencing was performed a chemogenomic assay of GEM using Saccharomyces cerevisiae as model cell and the results showed that more than 40% of genes related to GEM response in yeast possess unknown or dubious function. We choose two yeast mutants to individually validate the fitness defect results observed by chemogenomic assay, Δhmt1 and Δcsi1, and it was found that in addition to some already described pathways involved in GEM resistance, cells deficient in deneddylation enzyme Cop9 Signalosome Interactor 1 (Csi1p) presented a high sensitivity to GEM. This was confirmed by individual growth analyses of Δcsi1 cells exposed to GEM, and this phenotype was reverted with CSI1 complementation gene. Csi1p is a well-characterized homolog equivalent to human Csn6 subunit of COP9 signalosome (CSN) involved in deneddylation process. We highlighted too that epigenetic alterations, such as methylation mediated by protein arginine methyltransferase 1, play an important role in regulating gemcitabine treatment resistance. Our results point out new unexplored molecular pathways that can be used to overcome GEM resistance: the inhibition of CSN and the arginine methyltransferase activities.

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Funding

This research was supported by grants from State of São Paulo Research Foundation (FAPESP/Brazil), process number 2009/01303-1, 2015/07749-2 and 2011/04938-8. The authors also would like to thank the National Postdoctoral Program (PNPD/Capes - FCF-USP) [Award Number 1781837]. G. M. received a Productivity Fellowship from the Brazilian National Counsel of Technological and Scientific Development (CNPq 309595/2016-9).

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Correspondence to Gisele Monteiro.

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de Sousa Cavalcante, L., Costa-Silva, T.A., Souza, T.A. et al. Chemogenomic study of gemcitabine using Saccharomyces cerevisiae as model cell—molecular insights about chemoresistance. Braz J Microbiol 51, 489–496 (2020). https://doi.org/10.1007/s42770-019-00154-7

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Keywords

  • Gemcitabine
  • Pancreatic cancer
  • Next-generation sequencing
  • Arginine methyltransferase
  • Csn6 subunit of COP9 signalosome
  • Chemoresistance pathway