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Cytotoxic and antiproliferative effects induced by a non terpenoid polar extract of A. indica seeds on 3T6 murine fibroblasts in culture

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Abstract

Neem oil is a natural product obtained from the seeds of the tree Azadirachta indica. Its composition is very complex and the oil exhibits a number of biological activities. The most studied component is the terpenoid azadirachtin which is used for its insecticidal and putative antimicrobial properties. In this report we investigate the biological activity of partially purified components of the oil obtained from A. indica. We show that the semi-purified fractions have moderate to strong cytotoxicity. However, this is not attributable to azadirachtin but to other active compounds present in the mixture. Each fraction was further purified by appropriate extraction procedures and we observed a differential cytotoxicity in the various sub-fractions. This led us to investigate the mode of cell death. After treatment with the oil fractions we observed positivity to TUNEL staining and extensive internucleosomal DNA degradation both indicating apoptotic death. The anti-proliferative properties of the neem oil-derived compounds were also assayed by evaluation of the nuclear PCNA levels (Proliferating Cell Nuclear Antigen). PCNA is significantly reduced in cells treated with a specific fraction of neem oil. Finally, our results strongly suggest a possible involvement of the mitochondrial pathway in the apoptotic death.

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Authors and Affiliations

  1. Biotechnology Biological Control Agency, V. del Bosco, 10 – 00060 Sacrofano, Roma, Italy

    Vincenzo Di Ilio & Massimo Cristofaro

  2. Dip.to Genetica e Biologia Molecolare, Università di Roma “La Sapienza”, P.le A. Moro, 5 – 00185, Roma, Italy

    Nicoletta Pasquariello & Gianfranco Risuleo

  3. ENEA C.R. Casaccia, V. Anguillarese, S. M. Galeria, 301 – 00060, Roma, Italy

    Andrew S. van der Esch & Massimo Cristofaro

  4. Dip.to Biologia cellulare e dello Sviluppo, Università di Roma “La Sapienza”, P.le A. Moro, 5 – 00185, Roma, Italy

    Vincenzo Di Ilio, Gianfranco Scarsella & Gianfranco Risuleo

Authors
  1. Vincenzo Di Ilio
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  2. Nicoletta Pasquariello
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  3. Andrew S. van der Esch
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  4. Massimo Cristofaro
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  5. Gianfranco Scarsella
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  6. Gianfranco Risuleo
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Correspondence to Gianfranco Risuleo.

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Ilio, V.D., Pasquariello, N., Esch, A.S.v.d. et al. Cytotoxic and antiproliferative effects induced by a non terpenoid polar extract of A. indica seeds on 3T6 murine fibroblasts in culture. Mol Cell Biochem 287, 69–77 (2006). https://doi.org/10.1007/s11010-005-9062-x

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  • DOI: https://doi.org/10.1007/s11010-005-9062-x

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