A traditional poly herbal medicine “Le Pana Guliya” induces apoptosis in HepG₂ and HeLa cells but not in CC1 cells: an in vitro assessment

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• 作者：Nekadage Don Amal Wageesha ; Preethi Soysa ; Keerthi Atthanayake…
• 关键词：Anti ; cancer activity ; MTT assay ; LDH assay ; GSH ; Rhodamine123 ; Cytotoxicity
• 刊名：Chemistry Central Journal
• 出版年：2017
• 出版时间：December 2017
• 年：2017
• 卷：11
• 期：1
• 全文大小：1955KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry/Food Science, general;
• 出版者：Springer International Publishing
• ISSN：1752-153X
• 卷排序：11

文摘

“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 (HepG₂ and 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 HepG₂ and HeLa cells with an EC₅₀ value 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 EC₅₀ value 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.