Morphological Alteration and Intracellular ROS Generation Confirm Apoptosis Induction on EAC Cells by Leucas Indica Bark Extract

ABSTRACT

The phytochemical constituents, antioxidant, cytotoxicity and anti-cancer activities of Leucas indica bark extract in methanol were evaluated. Brine shrimp lethality assay was used to assess the cytotoxicity while antioxidant capability was determined by DPPH and H₂O₂ free radical scavenging assay. Cell growth inhibition was estimated through hemocytometric cell counting after 5 d of treatment using EAC cells in in vivo mice model. This inhibition was due to the induction of apoptosis observed through fluorescence microscopy. The extract showed anticancer activity against EAC cells with 58.27% cell growth inhibition at 100 mg kg⁻¹ b.w. Morphological alterations identified demonstrated apoptosis induction supported by intracellular ROS generation and DNA fragmentation.

KEYWORDS:

• Anticancer
• antioxidant
• DNA fragmentation

Abbreviations

DAPI: 4′,6-diamidino-2-phenylindole; DCFH-DA: 2’, 7 ’ - dichloro-dihydro-fluorescein diacetate DPPH: 1,1-diphenyl-2-picrylhydrazyl; EAC: Ehrlich ascites carcinoma; H₂O₂: Hydrogen peroxide; PBS: Phosphate buffer saline; ROS: Reactive oxygen species; SD: Standard Deviation

Acknowledgments

The authors thank all the individuals who helped in this research. We extend our acknowledgment towards the technical support provided by Molecular and Cellular biology laboratory, Molecular Biology and Protein Science Laboratory, University of Rajshahi, Nutrition and Food Technology and Chemical Engineering, Jashore University of Science and Technology.

Disclosure Statement

No potential conflict of interest was reported by the author(s).

Data Availability

All the data expressed in the present paper will be readily available upon request ([email protected]).

Additional information

Funding

This research did not receive any specific grant from funding agencies.