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Carbamazepine, a Histone Deacetylase Inhibitor Induces Apoptosis in Human Colon Adenocarcinoma Cell Line HT-29

  • Sohaib M
  • Ezhilarasan DEmail author
Original Research
  • 8 Downloads

Abstract

Background and Aim

Colon cancer ranks fourth and is responsible for causing 10% cancer-related mortality in western countries. Its incidence is rising in many countries due to widespread adoption of the Western diet and lifestyle. Carbamazepine (CBZ) is a FDA-approved antiepileptic drug and a histone deacetylase inhibitor. The aim of this study is to evaluate the cytotoxic potentials of CBZ in human colon cancer cells (HT-29 cells).

Methods

HT-29 cells were treated with 36 and 76 μg/ml of CBZ for 24 h. The cytotoxic effect was evaluated by MTT assay. The intracellular reactive oxygen species (ROS) expression was evaluated through dichloro-dihydro-fluorescein diacetate staining. Morphological changes related to apoptosis were evaluated by dual staining with acridine orange and ethidium bromide. Mitochondrial membrane potential was evaluated by rhodamine 123 staining. Immunofluorescence analysis of caspase 3 was done with confocal microscopy.

Results

CBZ caused significant cytotoxicity in HT-29 cells and the effect was concentration dependent. CBZ treatments also caused significant expression of ROS in HT-29 cells. Dual staining showed early and late apoptotic cells and morphological alterations induced by the CBZ. Confocal microscopic studies confirmed the increased caspase 3 expression in CBZ-treated cells.

Conclusion

CBZ induced apoptosis in HT-29 cell through ROS generation and caspase 3 expression and these results pave the way for further in vivo studies.

Keywords

Antiepileptics Apoptosis Colon cancer Caspase Cytotoxicity 

Notes

Acknowledgments

We thank Dr. R. Parameshwari, Researcher, Sri Ramachandra Institute of Higher Education and Research, Chennai, India for her scientific contribution to this work.

Authors’ Contributions

MS performed the research and written the manuscript; DE conceived the ideas, performed the research, and analyzed the data and corrected the manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pharmacology, Saveetha Dental CollegeSaveetha Institute of Medical and Technical Sciences (SIMATS)ChennaiIndia
  2. 2.Biomedical Research Unit and Laboratory Animal Centre, Saveetha Dental College (SDC)Saveetha Institute of Medical and Technical SciencesChennaiIndia

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