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Cationic Antimicrobial Peptides Derived from Crocodylus siamensis Leukocyte Extract, Revealing Anticancer Activity and Apoptotic Induction on Human Cervical Cancer Cells

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Abstract

Known antimicrobial peptides KT2 and RT2 as well as the novel RP9 derived from the leukocyte extract of the freshwater crocodile (Crocodylus siamensis) were used to evaluate the ability in killing human cervical cancer cells. RP9 in the extract was purified by a combination of anion exchange column and reversed-phase HPLC, and its sequence was analyzed by mass spectrometry. The novel peptide could inhibit Gram-negative Vibrio cholerae (clinical isolation) and Gram-positive Bacillus pumilus TISTR 905, and its MIC values were 61.2 µM. From scanning electron microscopy, the peptide was seen to affect bacterial surfaces directly. KT2 and RT2, which are designed antimicrobial peptides using the C. siamensis Leucrocin I template, as well as RP9 were chemically synthesized for investigation of anticancer activity. By Sulforhodamine B colorimetric assay, these antimicrobial peptides could inhibit both HeLa and CaSki cancer cell lines. The IC50 values of KT2 and RT2 for HeLa and CaSki cells showed 28.7–53.4 and 17.3–30.8 µM, while those of RP9 were 126.2 and 168.3 µM, respectively. Additionally, the best candidate peptides KT2 and RT2 were used to determine the apoptotic induction on cancer cells by human apoptosis array assay. As a result, KT2 and RT2 were observed to induce apoptotic cell death in HeLa cells. Therefore, these results indicate that KT2 and RT2 with antimicrobial activity have a highly potent ability to kill human cervical cancer cells.

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Abbreviations

ACN:

Acetonitrile

AMPs:

Antimicrobial peptides

hRBCs:

Human red blood cells

IC50 :

Half maximal inhibitory concentration

MIC:

Minimal inhibitory concentration

TFA:

Trifluoroacetic acid

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Acknowledgments

This work was financially supported by the Research and Researchers for Industries (RRI), the Thailand Research Fund, the National Research Council of Thailand, Grant no. 2556-22, and Sriracha Moda Co., Ltd., Chon Buri, Thailand. Additionally, we would like to thank the Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI) as well as the Department of Biochemistry, Faculty of Science, Khon Kaen University for essential support by laboratory facilities and giving much effective knowledge.

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Authors and Affiliations

  1. Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand

    Tinnakorn Theansungnoen, Surachai Maijaroen, Nisachon Jangpromma, Nualyai Yaraksa, Sakda Daduang & Sompong Klaynongsruang

  2. Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand

    Tinnakorn Theansungnoen, Surachai Maijaroen & Sompong Klaynongsruang

  3. Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand

    Nisachon Jangpromma

  4. Division of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Sakda Daduang

  5. Sriracha Moda Co., Ltd., Chon Buri, 20110, Thailand

    Theeranan Temsiripong

  6. Department of Clinical Chemistry, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Jureerut Daduang

Authors
  1. Tinnakorn Theansungnoen
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  2. Surachai Maijaroen
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  3. Nisachon Jangpromma
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  4. Nualyai Yaraksa
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  5. Sakda Daduang
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  7. Jureerut Daduang
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  8. Sompong Klaynongsruang
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Correspondence to Sompong Klaynongsruang.

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10930_2016_9662_MOESM2_ESM.tif

Inhibitory activity of Crocodylus siamensis leukocyte extract (6.82 mg mL−1) against bacteria by liquid growth inhibition assay. The mean with standard deviation (n = 3) was evaluated for all data (TIFF 9410 kb)

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Theansungnoen, T., Maijaroen, S., Jangpromma, N. et al. Cationic Antimicrobial Peptides Derived from Crocodylus siamensis Leukocyte Extract, Revealing Anticancer Activity and Apoptotic Induction on Human Cervical Cancer Cells. Protein J 35, 202–211 (2016). https://doi.org/10.1007/s10930-016-9662-1

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  • DOI: https://doi.org/10.1007/s10930-016-9662-1

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