Welcome to International Network for Natural Sciences | INNSpub

Induction of apoptosis by Ocimum sanctum (Holy basil) leaf extract against myeloid leukemia cells

Research Paper | August 1, 2020

| Download 19

Antara Haldar, Nilanjana Deb, Shila Elizabeth Besra

Key Words:

Int. J. Biosci.17( 2), 194-206, August 2020

DOI: http://dx.doi.org/10.12692/ijb/17.2.194-206


IJB 2020 [Generate Certificate]


Antineoplastic effect of Ocimum sanctum leaf extract was reported in our previous work. The cytotoxic potential of methanolic extract Ocimum sanctum (OSLE) was evaluated by MTT assay. Annexin-V FITC double staining was done to examine whether cell death occurred via apoptosis or necrosis. Gel electrophoresis, fluorescence microscopy, confocal microscopy and light microscopy were used to determine the cell morphology and fragmented DNA. JC-1 staining was also performed to measure the mitochondrial membrane potential. The phase distribution of the leukemia cell study was done by the cell cycle study. The pathway of apoptosis was studied by Caspase assay. OSLE significantly inhibited the cell viability in a time and concentration dependent manner in all the leukemia cells. After the treatment with OSLE, leukemia cells showed various signs of apoptosis like nuclear fragmentation, formation of apoptotic bodies, chromatin condensation and degraded DNA bands in cancer cells. The presence of apoptotic cells in the early and late stages was confirmed by the flow cytometric analysis. The cell cycle phase arrest was observed in the G0/G1 phase in the myeloid lymphoma and the multidrug resistant leukemia cells. A significant change in the mitochondrial membrane potential was observed in the OSLE treated leukemia cells with MMP shift assay. Caspase assay reveals that the pathway followed may be intrinsic apoptosis pathway. These finding suggest that the methanolic extract of Ocimum sanctum (OSLE) possesses anti-leukemic effects via apoptosis not necrosis along with insignificant toxicity.


Copyright © 2020
By Authors and International Network for
Natural Sciences (INNSPUB)
This article is published under the terms of the Creative
Commons Attribution Liscense 4.0

Induction of apoptosis by Ocimum sanctum (Holy basil) leaf extract against myeloid leukemia cells

Besra SE, Sharma RM, Gomes A. 1996. Anti-inflammatory effect of petroleum ether extract of leaves of Litchi chinensis Gaertn (Sapindaceae). Journal of Ethnopharmacol 54, 1-6. https://doi.org/ 10.1016/0378-8741(96)01440-7

Bindu J,  Narendhirakannan RT. 2019. Role of medicinal plants in the management of diabetes mellitus: a review, 3 Biotech 9(1), 4.

Bob L, Jacob M Rowe. 2016. Introduction to the review series on advances in acute myeloid leukemia. American society of Haematology. Blood 127, 1. https://doi:10.1182/blood-2015-10-662684

Cao Z, Li Y. 2002. Chemical induction of cellular antioxidants affords marked protection against oxidative injury in vascular smooth muscle cells. Biochemical and Biophysical Research Communications 292, 50-7.

Gao P, Seebacher NA, Hornicek F, Guo Z, Duan Z. 2018. Advances in Sarcoma Gene Mutations and Therapeutic Targets. Cancer Treatment Reviews 62, 98-109. https://doi.org/ 10.1016/j.ctrv. 2017.11.1

Gaurav A, Nilanjana D, Piyali D, Vishal S, Kanika K, Shila EB. 2019. Anti-Neoplastic effect of Ocimum Sanctum methanol extract on colorectal carcinoma, gastric carcinoma, hepatocellular carcinoma & normal cells. World Journal of Pharmaceutical Research 8, 984-1001.

Graça MD, Susan SD, Rochelle EC, Martha S Linet, Lindsay MM. 2012. Acute leukemia incidence and patient survival among children and adults in the United States, 2001-2007 Blood 119(1), 34-43. https://doi:10.1182/blood-2011-04-347872

Gupta SD, Debnath A, Saha A, Giri B, Tripathi G, Vedasiromoni JR, Antony G, Aparna G. 2007. Indian black scorpion (Heterometrus bengalensis Koch) venom induced antiproliferative and apoptogenic activity against human leukemic cell lines U937 and K562. Leuk Research 31, 817-25. https://doi:10.1016/j.leukres.2006.06.004

Hanahan D, Weinberg RA. 2011. Hallmarks of cancer: The next generation. Cell 144(5), 646-674. https://doi:10.1016/j.cell.2011.02.013

Herrmann H, Lorenz HM, Voll R, Grunke M, Woith W, Kalden JR. 1994. A rapid and simple method for isolation of apoptotic DNA fragments. Nucleic Acids Research 22(24), 5506-7. https://doi:10.1093/nar/22.24.5506

Houghton P, Fang R, Techatanawat I, Steventon G, Hylands PJ, Lee CC. 2007. The sulphorhodamine (SRB) assay and other approaches to testing plant extracts and derived compounds for activities related to reputed anticancer activity. Methods 42(4), 377-87. https://doi:10.1016 /j. ymeth. 2007.01.003

Karthikeyan K, Gunasekaran P, Ramamurthy N, Govindasamy S. 1999. Anticanceractivity of Ocimum sanctum. Pharmaceutical Biology 37(4), 285-290. https://doi.org/10.1076/i.

Lam SN, Neda GD, Rabeta MS. 2017. The anticancer effect of Ocimum tenuiflorum leaves. Food Research 2 (2), 154-162. https://doi: 10.26656/fr. 2017.2(2).251

Martins E. 2013. The growing use of herbal medicines: issues relating to adverse reactions and challenges in monitoring safety. Frontiers in Pharmacology 4, 177. https://doi:10.3389/fphar. 2013.00177

Mishell BB, Shiigi SM, Henry C. 1980. Preparation of mouse cell Suspensions. In: Mishell BB, Shiigi SM, Eds. Selected Methods in Cellular Immunology 23. San Francisco. Freeman WH & Co. pp. 21.

Mohammadian M, Pakzad R, Mohammadian AH, Salehiniya H. 2018. A study on the incidence and mortality of leukemia and their association with the human development index (HDI) worldwide in 2012. World Cancer Research Journal 5(2), e1080.

Murmu N, Mitra S, Das M, Gomes A, Vedasiromoni JR, Ghosh M, Bhattacharya M, Ghosh P, Biswas J, Bhattacharya S, Sur P. 2001. Boron compounds against human leukemic cells. Journal of Experimental & Clincal Cancer Research 20, 511-5.

Nilanjana D, Anita H, Soumyasree D, Ashok P, Shila EB. 2017. Lawsonia Alba leaves induce apoptosis and cell cycle arrest in B16F10 melanoma cells. International Journal of Pharmacy and Pharmaceutical Sciences 10(5), 96-101. https:// doi:10.22159/ijpps.2018v10i5.24526

Piyali B, Anupam B. 2013. Ocimum sanctum Linn. (Tulsi): An ethno medicinal plant for the prevention and treatment of cancer. Anti-Cancer Drugs 24(7), 659-666. https://doi:10.1097/cad.0b013e328361aca1

Priyabrata P, Pritishova B, Debajyoti D, Sangram K Panda. 2010. Ocimum sanctum Linn. A reservoir plant for therapeutic applications. Pharmacognosy Review 4(7), 95-105. https://doi:  10.4103/0973-7847.65323

Roy S, Besra SE, De T, Banerjee B, Mukherjee J, Vedasiromoni JR. 2008. Induction of Apoptosis in Human Leukemic Cell Lines U937, K562 and HL-60 by Litchi chinensis Leaf Extract Via Activation of Mitochondria Mediated Caspase Cascades. The Open Leukemia Journal 1, 14. https://doi: 10.2174/ 18768 16400901010001

Shamik B, Tanaya D, Aparna G, Archita B, Antony G, Sandhya RD. 2013. A cytotoxic protein (BF-CT1) purified from Bungarus fasciatus venom acts through apoptosis, modulation of PI3K/AKT, MAPKinase pathway and cell cycle regulation. Toxicon74, 138-150. https://doi: 10.1016/j.toxicon .2013.08.052

Sheikh A, Yalavarthy PD, Adepelly U. 2016. Plants as Potential Resources of Anti-cancer Drugs. Current Trends in Biotechnology and Pharmacy 10(1), 92-107. https:// doi:10.1038/srep25462

Singh N, Verma P, Pandey BR, Bhalla M. 2012.Therapeutic Potential of Ocimum sanctum in Prevention and Treatment of Cancer and Exposure to Radiation. International Journal of Pharmaceutical Sciences and Drug Research 4(2), 97-104.

Siti MA, Ibrahim J, Md Areeful H, Laiba A. 2018. Exploring the Leaves of Annona muricata L. as a Source of Potential Anti-inflammatory and Anticancer Agents. Frontiers in Pharmacology 9, 661. https://doi: 10.3389/fphar.2018.00661

Sridevi M, Bright J, Yamini K. 2016. Anti-Cancer effect of Ocimum sanctum ethanolic extract in non-small cell lung carcinoma cell line. International Journal of Pharmacy and Pharmaceutical Sciences 8(4), 975-1491.

Uma P, Devi. 2001. Radio-protective, anti-carcinogenic and anti-oxidant properties of the Indian Holy Basil, Ocimum sanctum (Tulasi). Indian Journal of Experimental Biology 39, 185-190.

Xianwen Y, Molly P, Man W, Ray Kit Ng. 2019. Aberrant DNA Methylation in Acute Myeloid Leukemia and Its Clinical Implications. International Journal of Molecular Scienes 20(18), 4576. https:// doi: 10.3390/ijms20184576