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Licensed Unlicensed Requires Authentication Published by De Gruyter March 17, 2017

ROS-mediated induction of apoptosis by benzoquinone embelin in human colon adenocarcinoma cells HT-29

  • Kodandaram Sumalatha , Mohan Gowda and Sreepriya Meenakshisundaram EMAIL logo

Abstract

Background

Embelin is a benzoquinone reported to possess anticancer activity in several in vivo and in vitro models of carcinogenesis, especially hematopoietic and prostate malignancy. A detailed investigation on the influence of embelin on epithelial malignancy model system, especially colon adenocarcinoma, is lacking. The objective of the current study is to investigate the antiproliferative, antiinvasive and proapoptotic potential of embelin on colon adenocarcinoma cell line HT-29.

Methods

The effect of embelin (35 µg/mL for 24 h) on cell proliferation was assessed by Sulforhodamine B assay and bromodeoxyuridine incorporation test, antiinvasive effect by Boyden chamber assay and scratch assay. Proapoptotic effects of embelin were determined by studies on DNA fragmentation, annexin V-FITC labeling, TUNEL assay, COMET assay and assay of caspase-3 activity. Influence of embelin on the expression of genes regulating apoptosis (caspase 3 and 9, Bcl-2, Bax, cytochrome C and X-linked inhibitor of apoptosis protein) and migration/invasion (matrix metalloproteinase [MMP]-2 and MMP-9) was investigated by reverse transcription polymerase chain reaction (PCR). Further, the effect of embelin on the levels of reactive oxygen species (ROS), lipid peroxides, nitric oxide, mitochondrial membrane potential and antioxidant status (total reduced glutathione [GSH] and GSH-S-transferase) was evaluated.

Results

Results implicated that embelin treatment inhibited proliferation (IC50 35 µg/mL), induced DNA fragmentation, phosphatidyl serine externalization, increased caspase expression, decreased cell migration and expression of MMPs in HT-29 cells. Interestingly, embelin exhibited prooxidant effect on HT-29 cells and induced excessive ROS generation resulting in apoptotic cell death.

Conclusions

To conclude, embelin treatment could be a promising strategy for the chemotherapy of colon cancer.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: The financial assistance provided to this work by the Department of Science and Technology – Science and Engineering Research Board (DST-SERB No: SR/FT/LS-120/2008), Government of India to Dr M. Sreepriya under the Fast track scheme for young scientists is gratefully acknowledged.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis and interpretation of data; in the writing of the report or in the decision to submit the report for publication.

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Received: 2016-10-28
Accepted: 2017-1-26
Published Online: 2017-3-17
Published in Print: 2017-3-16

© 2017 Walter de Gruyter GmbH, Berlin/Boston

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