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Apoptogenic effects of Tricholoma giganteum on Ehrlich’s ascites carcinoma cell

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Abstract

This study explored the efficacy of Fa fraction of Tricholoma giganteum against Ehrlich’s ascites carcinoma (EAC). Mechanisms of apoptogenic effect of the fraction were delineated. The flow cytometric analysis of EAC cells, showed an increase in number of cells in sub-G0/G1 population and reduction in the G2/M phase due to the treatment thus suggesting apoptosis. The induction of apoptosis has also been confirmed by nuclear staining that demonstrated distinctive morphological features of apoptosis. Our data also revealed an increase in the expression of pro-apoptotic protein p53 in EAC and induced factors contributing to apoptosis. Pro-apoptotic gene Bax was up-regulated during p53-mediated apoptosis. No significant change in the expression of anti-apoptotic protein Bcl-2 was observed ensuing in decrease of the Bcl-2/Bax ratio. p53-mediated growth arrest involves p21 as a major effecter, which interestingly showed moderate elevation. All these observations indicate that Fa fraction of T. giganteum induces apoptogenic signal in EAC.

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Abbreviations

AP:

Alkaline phosphatase

BCIP:

Bromochloroindolyl phosphate

DAPI:

4′,6′-Diamidino-2-phenylindole

dNTP:

Deoxynucleoside triphosphates

DTT:

Dithiothreitol

EAC:

Ehrlich’s ascites carcinoma

FACS:

Fluorescence-activated cell sorter

FITC:

Fluorescein isothiocyanate

NBT:

Nitrobluetetrazolium

PBS:

Phosphate buffered saline,

PI:

Propidium iodide

RT-PCR:

Reverse transcription-polymerase chain reaction

SDS:

Sodium dodecyl sulfate

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Acknowledgments

The financial support of the University Grants Commission-Research Fellowship in Science for Meritorious Students (UGC-RFSMS), New Delhi, is gratefully acknowledged.

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Authors and Affiliations

  1. Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, Kolkata, 700019, India

    Soumya Chatterjee, Gunjan Biswas, Swarnendu Chandra & Krishnendu Acharya

  2. Entomology and Wildlife Biology Research Laboratory, Department of Zoology, University of Calcutta, Kolkata, 700019, India

    Goutam Kumar Saha

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  1. Soumya Chatterjee
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  2. Gunjan Biswas
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  3. Swarnendu Chandra
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  4. Goutam Kumar Saha
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  5. Krishnendu Acharya
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Correspondence to Krishnendu Acharya.

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Chatterjee, S., Biswas, G., Chandra, S. et al. Apoptogenic effects of Tricholoma giganteum on Ehrlich’s ascites carcinoma cell. Bioprocess Biosyst Eng 36, 101–107 (2013). https://doi.org/10.1007/s00449-012-0765-6

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  • DOI: https://doi.org/10.1007/s00449-012-0765-6

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