APOPTOSIS-INDUCING POTENTIAL OF LAWSONIA ALBA LAM. LEAVES ON HEPATOCELLULAR CARCINOMA (HEP-G2) CELLS ALONG WITH ITS ANTI-OXIDANT PROPERTY
Objective: The current study investigated the anti-cancer potential of methanolic and ethyl acetate fraction of Lawsonia alba L. (Lythraceae) leaves extract on Hep-G2 and RAW 264.7 cells along with in vitro anti-oxidant property of the ethyl acetate fraction.
Methods: The cytotoxic activity of methanolic extract and its fractions had been studied by MTT assay on Hep-G2 and RAW 264.7 cells. Morphological study of Hep-G2 cells was performed by light, fluorescence and confocal microscope. 1% agarose gel electrophoresis, detection of apoptosis and cell cycle arrest by flow cytometric analysis had been performed to determine the proportion and stages of cellular apoptosis of Hep-G2 cells. In vitro anti-oxidant study of various fractions of MLA were performed by DPPH and Hydroxyl radical scavenging assay.
Results: Cytotoxicity study of MLA and ELA had been confirmed by MTT assay and the IC50 value were calculated to be 75.85Î¼g/ml and 32.81Î¼g/ml on Hep-G2 cell line respectively. Morphological study showed the arrays of nuclear changes including chromatin condensation and apoptotic body formation indicating that treatment with ELA, causes apoptotic changes in the hepatoma cells compared to the untreated control. Agarose gel electrophoresis study showed fragmented DNA in the form of a ladder. Flow cytometric analysis showed an appreciable number of cells in early apoptosis stage. The cells were arrested, mostly in G0/G1 phase of the cell cycle. Antioxidant property of ELA fraction was confirmed by free radical scavenging activity.
Conclusion: Ethyl acetate fraction of Lawsonia alba L. leaves possess potent apoptotic activity against Hep-G2 cell line along with notable anti-oxidant activity.
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