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In Vitro Cytotoxicity of Parasporins from Native Algerian Bacillus thuringiensis Strains Against Laryngeal and Alveolar Cancers

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Abstract

Parasporins (PS), a class of non-insecticidal and non-hemolytic crystal proteins of Bacillus thuringiensis (Bt), are being explored as promising anti-cancer agents due to their specific toxicity to cancer cells. This work is considered as a first initiative aiming at investigating Algerian soil Bt isolates’ activity and cytotoxic potential against cancer cells. A total of 48 Bacillus spp. were isolated from different sites in Algeria. Phenotypic and biochemical tests, 16S rDNA molecular identification, and microscopic observation of crystal have confirmed the identification of Bt for ten strains. A screening for non-hemolytic crystalline proteins was performed. Extraction, purification, and activation of non-hemolytic proteins by chromatographic analysis yielded several polypeptides of different molecular weights. A purified PS1, with pro-protein of 81 kDa and several peptides with different molecular weights (18–58 kDa) after activation by trypsin, has been identified from the strain BDzG. The NH2-terminal sequence deciphered in BLAST analysis showed homology to a Bt PS1 protein. Moreover, the screening of parasporin-1 (PS1) gene has also been performed. Cytocidal activity against human epithelial type 2 (HEp2) cells, considered to originate from a human laryngeal carcinoma, was observed with an IC50 equal to 2.33 μg/ml, while moderate cytotoxicity against adenocarcinomic human alveolar basal epithelial (A549) cells has been shown with IC50 equal to 18.54 μg/ml. No cytotoxicity against normal cells was noted. Fluorescence microscopy revealed a condensed or fragmented chromatin indicating the apoptotic death of HEp2 cells. Thus, Bt PS-producer isolated from Algerian soil might have a potential to join the arsenal of natural anti-cancer drugs with high therapeutic potential.

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Acknowledgements

The authors would like to express their thanks and gratitude to Mr. A. Adjerad for the statistical part assistance. Special thanks are also due to Mr. A.S. Moulla for constructively proofreading and language polishing services. This work is part of a Doctoral Thesis in Environmental Engineering by Mrs Lila Aberkane at the National Polytechnic School of Algiers (ENP, Algeria).

Funding

This study was financially supported by Nuclear Research Center of Algiers (CRNA). The funding entity had no contribution in the study design and data collection, analysis, and interpretation as well as the manuscript writing.

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Authors and Affiliations

  1. Medical Applications Department, Nuclear Applications Division, Nuclear Research Center of Algiers (CRNA), 02 Bd Frantz Fanon, P.O. Box 399, 16000, Algiers-Gare, Algiers, Algeria

    Lila Aberkane, Assia Nacer-Khodja, Abdelhakim Ouafek & Assia Djefal

  2. Laboratory of Cellular and Molecular Biology (LCMB), Microbiology Team, Faculty of Biological Sciences, University of Sciences and Technology of Houari Boumediene (USTHB), P.O. Box 32, El Alia, 16111, Bab Ezzouar, Algiers, Algeria

    Zahia Djenane, Lydia Neila Djouadi & Farida Nateche

  3. Laboratory of Bioactive Substances, Center of Biotechnology of Borj Cedria (CBBC), University of Tunis El Manar, P.O. Box 901, 2050, Hammam-Lif, Tunisia

    Lamjed Bouslama

  4. Environmental Engineering Department, National Polytechnic School of Algiers (ENP), 10 Rue des Frères Oudek, Hacène Badi, PO Box 182, 16200, El Harrach, Algiers, Algeria

    Lila Aberkane, Hocine Grib & Nabil Mameri

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  1. Lila Aberkane
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Correspondence to Assia Djefal.

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Ethical clearance and approval for the study was obtained from the Ethical Committee in Human Clinical Research [Mustapha PACHA University Hospital Centre of Algiers (CHUMA) and Pasteur Institute of Algiers (IPA), Algeria].

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Aberkane, L., Nacer-Khodja, A., Djenane, Z. et al. In Vitro Cytotoxicity of Parasporins from Native Algerian Bacillus thuringiensis Strains Against Laryngeal and Alveolar Cancers. Curr Microbiol 77, 405–414 (2020). https://doi.org/10.1007/s00284-019-01841-2

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