Abstract
Background
The in vivo anticancer effect of the Trema orientalis leaves crude methanol extract (TLME) was screened against Ehrlich ascites carcinoma (EAC) in Swiss albino mice.
Materials and methods
The cytotoxic activity of TLME was determined in vitro by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The growth inhibitory activity and morphological alterations were determined by the hemocytometer counting of the EAC cells using trypan blue dye. The apoptotic cells were assessed by DAPI (4′,6-diamidino-2-phenylindole) staining. The hematological and biochemical parameters of experimental mice were also estimated.
Results
After treatment with the TLME, the viable tumor cell count, morphological changes and nuclear damages of the EAC cells were observed along with the hematological parameters of the experimental mice. The LD50 of TLME was 3120.650 mg/kg body weight, and this extract was proven to be safe at a dose of as high as 800 mg/kg body weight. The oral administration of the TLME at 400 mg/kg body weight resulted in approximately 59% tumor cell growth inhibition compared with the control mice, with considerable apoptotic features, including membrane blebbing, chromatin condensation, nuclear fragmentation and aggregation of the apoptotic bodies in DAPI staining under a fluorescence microscope. The TLME also dose-dependently restored the altered hematological parameters to approximately normal levels. The TLME exhibited bolstering cytotoxic effect against the EAC cell with the IC50 value of 29.952 ± 1.816 μg/mL.
Conclusion
The TLME has potential as a natural anti-cancer product with apoptosis induction property and cytotoxicity against carcinoma cells.
Award Identifier / Grant number: 39.00.0000.012.02.013.17-170
Funding statement: This research was funded by the National Science and Technology (NST) Grant of the Ministry of Science and Technology of the People’s Republic of Bangladesh (Grant No: 39.00.0000.012.02.013.17-170).
Acknowledgments
The authors are thankful to Dr. Md. Abu Reza, Professor, Protein Science Laboratory, Department of Genetic Engineering and Biotechnology, University of Rajshahi, Bangladesh for the technical and laboratory support provided.
Author contributions: MMR and SA designed the project and experiments; MMR and SA analyzed the results; MMK, MAAN and MMH performed the experimental work; MMR, BKD and SA wrote the manuscript; and MMR supervised the project. All the authors have read and approved the final manuscript.
Competing interests: The authors state no conflict of interest.
Ethical approval: The methodology used in this work and the handling of the experimental animals were approved by the Institutional Animal, Medical Ethics, Biosafety and Biosecurity Committee (IAMEBBC) for Experimentations on Animal, Human, Microbes and Living Natural Sources (License No: 31/320/IAMEBBC/IBSC), Institute of Biological Sciences, University of Rajshahi, Bangladesh.
Specimen voucher number for the plant tested: National Herbarium, Dhaka, Bangladesh; Voucher number: DACB/42864.
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