Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 390, Issue 10, pp 1005–1013 | Cite as

Cytotoxicity against tumor cell lines and anti-inflammatory properties of chitinases from Calotropis procera latex

  • Carolina Araújo Viana
  • Márcio V. Ramos
  • José Delano Barreto Marinho Filho
  • Letícia Veras Costa Lotufo
  • Ingrid Samantha Tavares Figueiredo
  • Jefferson Soares de Oliveira
  • Pietro Mastroeni
  • José Vitor Lima-FilhoEmail author
  • Nylane Maria Nunes AlencarEmail author
Original Article


The role of chitinases from the latex of medicinal shrub Calotropis procera on viability of tumor cell lines and inflammation was investigated. Soluble latex proteins were fractionated in a CM Sepharose Fast-Flow Column and the major peak (LPp1) subjected to ion exchange chromatography using a Mono-Q column coupled to an FPLC system. In a first series of experiments, immortalized macrophages were cultured with LPp1 for 24 h. Then, cytotoxicity of chitinase isoforms (LPp1-P1 to P6) was evaluated against HCT-116 (colon carcinoma), OVCAR-8 (ovarian carcinoma), and SF-295 (glioblastoma) tumor cell lines in 96-well plates. Cytotoxic chitinases had its anti-inflammatory potential assessed through the mouse peritonitis model. We have shown that LPp1 was not toxic to macrophages at dosages lower than 125 μg/mL but induced high messenger RNA expression of IL-6, IL1-β, TNF-α, and iNOs. On the other hand, chitinase isoform LPp1-P4 retained all LPp1 cytotoxic activities against the tumor cell lines with IC50 ranging from 1.2 to 2.9 μg/mL. The intravenous administration of LPp1-P4 to mouse impaired neutrophil infiltration into the peritoneal cavity induced by carrageenan. Although the contents of pro-inflammatory cytokines IL-6, TNF-α, and IL1-β were high in the bloodstreams, such effect was reverted by administration of iNOs inhibitors NG-nitro-L-arginine methyl ester and aminoguanidine. We conclude that chitinase isoform LPp1-P4 was highly cytotoxic to tumor cell lines and capable to reduce inflammation by an iNOs-derived NO mechanism.


Anticancer activity Folk medicine Laticifer proteins Nitric oxide 



This study is part of the consortium Molecular Biotechnology of Plant Latex supported by the Northeast Biotechnology Network (RENORBIO-Brazil) and funded by the Brazilian National Counsel of Technological and Scientific Development (CNPq). The authors are also grateful to CAPES for scholarship support to Carolina Viana, and Ceará State Foundation (FUNCAP—Program PPSUS). We thank Panagiotis Tourlomousis, Kate Fitzgerald, and Clare Bryant for providing the macrophage cell lineage used in the present study.


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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Carolina Araújo Viana
    • 1
  • Márcio V. Ramos
    • 1
  • José Delano Barreto Marinho Filho
    • 2
  • Letícia Veras Costa Lotufo
    • 3
  • Ingrid Samantha Tavares Figueiredo
    • 4
  • Jefferson Soares de Oliveira
    • 5
  • Pietro Mastroeni
    • 6
  • José Vitor Lima-Filho
    • 7
    • 8
    Email author
  • Nylane Maria Nunes Alencar
    • 4
    • 9
    Email author
  1. 1.Department of Biochemistry and Molecular BiologyFederal University of CearáFortalezaBrazil
  2. 2.Medical SchoolFederal University of PiauíParnaíbaBrazil
  3. 3.Department of PharmacologyUniversity of São PauloSão PauloBrazil
  4. 4.Department of Physiology and PharmacologyFederal University of CearáFortalezaBrazil
  5. 5.Biomedicine SchoolFederal University of PiauíParnaíbaBrazil
  6. 6.Department of Veterinary MedicineUniversity of CambridgeCambridgeUK
  7. 7.Department of BiologyFederal Rural University of PernambucoRecifeBrazil
  8. 8.Departamento de Biologia, Laboratório de Microbiologia e ImunologiaUniversidade Federal Rural de PernambucoRecifeBrazil
  9. 9.Departamento de Fisiologia e FarmacologiaUniversidade Federal do CearáFortalezaBrazil

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