APOPTOSIS-INDUCING POTENTIAL OF LAWSONIA ALBA LAM. LEAVES ON HEPATOCELLULAR CARCINOMA (HEP-G2) CELLS ALONG WITH ITS ANTI-OXIDANT PROPERTY

Soumyasree Dutta; Nilanjana Deb; Ashok Kumar Pattnaik; Shila Elizabeth Besra

doi:10.22159/ijpps.2016.v8i9.12815

Authors

Soumyasree Dutta Cancer Biology and Inflammatory Disorder Division, CSIR- Indian Institute of Chemical Biology, 4,Raja S.C. Mullick Road, Kolkata 700032,West Bengal, India
Nilanjana Deb Cancer Biology and Inflammatory Disorder Division, CSIR- Indian Institute of Chemical Biology, 4,Raja S.C. Mullick Road, Kolkata 700032,West Bengal, India
Ashok Kumar Pattnaik Department of Pharmaceutical Science and Technology, Birla Institute of Technology Mesra, Ranchi, 835215, Jharkhand, India
Shila Elizabeth Besra Cancer Biology and Inflammatory Disorder Division, CSIR- Indian Institute of Chemical Biology, 4,Raja S.C. Mullick Road, Kolkata 700032,West Bengal, India

DOI:

https://doi.org/10.22159/ijpps.2016.v8i9.12815

Keywords:

Lawsonia alba, Leaves, Hepatocellular carcinoma, Apoptosis, Antioxidant

Abstract

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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