Touseef Rehan, David MacEwan, Nasrullah Shah, Tabassum Rehan, Riffat Tahira, Sheeba Murad, Mariam Anees, Iram Murtaza, Muhammad Farman, Obaid-ur-Rahman Abid and Aneesa Sultan* Pages 3681 - 3691 ( 11 )
Purpose: Leukemia, one of the major cancers, affects a large proportion of people around the world. Better treatment options for leukemia are required due to a large number of side effects associated with current therapeutic regimens. In the present study, we sought to determine the pathway of triggering apoptosis of leukemic cells by Ocimum basilicum (O. basilicum) plant extract.Materials/Methods: Methanolic extract of the O. basilicum plant material was prepared. The crude extract was fractionated into several fractions through column chromatography using ethyl acetate and n-hexane as eluting solvents. Cell viability of leukemic cells was assessed via Cell titer GLO assay and apoptosis was measured through Annexin V/PI staining. Two apoptotic molecules JNK and caspases were analyzed through western blotting while pro-inflammatory cytokines TNFα, CCL2 and CXCL8 using qPCR. Fractions were characterized through LC-MS. Results: The most potent with lowest IC50 values among the fractions were BF2 (2:8 n-hexane:ethyl acetate) and BF3 (3:7 n-hexane:ethyl acetate). Cytotoxicity was associated with apoptosis. Apoptosis was found caspasedependent and P-JNK activation was detected sustained. A significant increase in the level of TNF α and a decrease in the level of CXCL8 were observed in BF2 and BF3 treated cells. Conclusion: The fractions of O. basilicum extract were found to kill cells following JNK pathway activation. Excellent results were obtained with BF2 and BF3 probably due to predominant Epicatechin and Cinnamic acid derivatives in these fractions.
Apoptosis, cytokines, cytotoxicity, leukemia, natural products, Ocimum basilicum.
Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Department of Translational Medicine, Faculty of Health Sciences, University of Liverpool, Liverpool L69 3BX, Department of Chemistry, Faculty of Chemical and Life Sciences, Abdul Wali Khan University, Mardan, Department of Biotechnology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Plant Genetic Resources Program, National Agricultural Research Centre, Park Road, Islamabad, Institute for Infection and Immunity, St George's University of London, London, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Department of Chemistry, Faculty of Natural Sciences, Quaid-i-Azam University, Islamabad, Department of Chemistry, Hazara University, Mansehra, 21120, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad