title: A traditional poly herbal medicine "Le Pana Guliya"induces apoptosis in HepG2and HeLa cells but not in CC1 cells: an in vitro assessment.
abstract: "Le Pana Guliya" (LPG) is a polyherbal formulation which is used to treat different types of cancers in traditional medicine. In this study we describe in vitro efficacy and mechanism of action of LPG on two cancer cell lines (HepG2and HeLa) compared with a normal cell line CC1. The MTT, LDH assays and protein synthesis were used to study antiproliferative activity of LPG while NO synthesis and GSH content were assayed to determine the oxidative stress exerted by LPG. Rhodamine 123 staining, caspase 3 activity, DNA fragmentation and microscopic examination of cells stained with ethidium bromide/acridine orange were used to identify the apoptosis mechanisms associated with LPG. The LPG showed the most potent antiproliferative effect against the proliferation of HepG2and HeLa cells with an EC50value of 2.72 ± 1.36 and 19.03 ± 2.63 µg/mL for MTT assay after 24 h treatment respectively. In contrast, CC1 cells showed an EC50value of 213.07 ± 7.71 µg/mL. Similar results were observed for LDH release. A dose dependent decrease in protein synthesis was shown in both cancer cell types compared to CC1 cells. The reduction of GSH content and elevation of cell survival with exogenous GSH prove that the LPG act via induction of oxidative stress. LPG also stimulates the production of NO and mediates oxidative stress. Rhodamine 123 assay shows the mitochondrial involvement in cell death by depletion of Δψ inducing downstream events in apoptosis. This results in increase in caspase-3 activity eventually DNA fragmentation and LPG induced apoptotic cell death. In conclusion the present study suggested that the LPG exerted an anticancer activity via oxidative stress dependent apoptosis. Therefore present study provides the scientific proof of the traditional knowledge in using LPG as an anticancer agent.
authors.affiliation: Department of Biochemistry and Chemistry, Faculty of Medicine, South Asian Institute of Technology and Medicine, Malabe, Sri Lanka ; Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka.
authors.forename: Nekadage Don Amal
authors.initials: ND
authors.lastname: Wageesha
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka.
authors.forename: Preethi
authors.initials: P
authors.lastname: Soysa
authors.affiliation: Department of Biochemistry and Chemistry, Faculty of Medicine, South Asian Institute of Technology and Medicine, Malabe, Sri Lanka.
authors.forename: Keerthi
authors.initials: K
authors.lastname: Atthanayake
authors.affiliation: Hussain Ebrahim Jamal Research Institute of Chemistry, Karachi, Pakistan ; Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Muhammad Iqbal
authors.initials: MI
authors.lastname: Choudhary
authors.affiliation: No: 9, Moragahapitiya, Balagola, Kengalle, Kandy, Sri Lanka.
authors.forename: Mahinda
authors.initials: M
authors.lastname: Ekanayake
keywords: Anti-cancer activity; Cytotoxicity; GSH; LDH assay; MTT assay; Rhodamine123