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

  • 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

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 75.85μg/ml and 32.81μg/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.

Keywords: Lawsonia alba, Leaves, Hepatocellular carcinoma, Apoptosis, Antioxidant

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References

1. Roopashree R, Mohan CD, Swaroop TR, Jagadish S, Rangappa KS. Synthesis characterization and in vivo biological evaluation of novel benzimidazoles as potential anticancer agents. Asian J Pharm Clin Res 2014;7:309-13.
2. Block TM, Mehta AS, Fimmel CJ, Jordan R. Molecular viral oncology of hepatocellular carcinoma. Oncogene 2003;22:5093-107.
3. Mikhaeil BR, Badria FA, Maatooq GT, Amer MM. Antioxidant and immunomodulatory constituents of henna leave. Z Naturforsch C: Biochem Biophys Biol Virol 2004;59:468-76.
4. Arun P, Purushotham KG, Jayarani JJ, D Kumari V. In vitro antibacterial activity and flavonoid contents of Lawsonia inermis (Henna). Int J PharmTech Res 2010;2:1178-81.
5. Uddin N, Siddiqui BS, Begum S. Chemical constituents and bioactivities of Lawsonia alba Lam. (Henna). J Chem Soc Pak 2013;35:476-85.
6. Bhandarkar M, Khan A. Protective effect of Lawsonia alba Lam. against CCl4 induced hepatic damage in albino rats. Indian J Exp Biol 2003;41:85-7.
7. Babili FE, Valentin A, Chatelain C. Lawsonia inermis: its anatomy and its antimalarial, antioxidant and human breast cancer cells MCF7 activities. Pharm Anal Acta 2013;4:203.
8. Uddin N, Siddiqui BS, Begum S, Bhatti HA, Khan A, Parveen S, et al. Bioactive flavonoids from the leaves of Lawsonia alba (Henna). Phytochem Lett 2011;4:454-8.
9. Kapadia GJ, Rao GS, Sridhar R, Ichiishi E, Takasaki M, Suzuki N, et al. Chemoprevention of skin cancer: effect of Lawsonia inermis L. (Henna) leaf powder and its pigment artifact, lawsone in the epstein-barr virus early antigen activation assay and in two-stage mouse skin carcinogenesis models. Adv Anticancer Agents Med Chem 2013;13:1500-7.
10. Ali NA, Julich WD, Kusnick C, Lindequist U. Screening of yemeni medicinal plants for antibacterial and cytotoxic activities. J Ethnopharmacol 2001;74:173-9.
11. Gupta SD, Debnath A, Saha A, Giri B, Tripathi G, Vedasiromoni JR, et al. Indian black scorpion (Heterometrus bengalensis Koch) venom induced antiproliferative and apoptogenic activity against human leukemic cell lines U937 and K562. Leuk Res 2007;31:817–25.
12. Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy. 42nd ed. Nirali Prakashan; 2008. p. A1-A6.
13. Surveswaran S, Cai YZ, Corke H, Sun M. Systematic evaluation of natural phenolic antioxidants from 133 Indian medicinal plants. Food Chem 2007;102:938–53.
14. Saeed SMG, Sayeed SA, Ashraf S, Naz S, Siddiqi R, Ali R, et al. A new method for the isolation and purification of lawsone from Lawsonia inermis and its ROS inhibitory activity. Pak J Bot 2013;45:1431-6.
15. Narayan S, Mittal A. In vitro antioxidant activity and free radical scavenging potential of roots of red sage. Asian J Pharm Clin Res 2015;8:417-21.
16. Cao Z, Li Y. Chemical induction of cellular antioxidants affords marked protection against oxidative injury in vascular smooth muscle cells. Biochem Biophys Res Commun 2002;292:50–7.
17. Herrmann H, Lorenz HM, Voll R, Grunke M, Woith W, Kalden JR. A rapid and simple method for isolation of apoptotic DNA fragments. Nucleic Acids Res 1994;22:5506-7.
18. Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labeled Annexin V. J Immunol Methods 1995;184:39–51.
19. Jacobson MD, Weil Miguel, Raff MC. Programmed cell death in animal development. Cell 1997;88:347-54.
20. Langman MJ, Cheng KK, Gilman EA, Lancashire RJ. Effect of anti-inflammatory drugs on overall risk of common cancer: a case-control study in general practice research database. Br Med J 2000;320:1642-6.
21. Lam MH, Liu Q, Elledge SJ, Rosen JM. Chk1 is haploinsufficient for multiple functions critical to tumor suppression. Cancer Cell 2004;6:45–59.
22. Martin SJ, Reutelingsperger CP, Mc Gahon AJ, Rader JA, van Schie RC, La Face DM, et al. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med 1995;182:1545-56.
23. Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P, et al. Features of apoptotic cells measured by flow cytometry. Cytometry 1992;13:795-808.
24. Darzynkiewicz Z, Bedner E, Smolewski P. Flow cytometry in analysis of cell cycle and apoptosis. Semin Hematol 2001;38:179-93.
25. Wising C, Azem J, Zetterberg M, Svensson LA, Ahlman K, Lagergard T. Induction of apoptosis/necrosis in various human cell lineages by Haemophilus ducreyi cytolethal distending toxin. Toxicon 2005;45:767-76.
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Dutta, S., N. Deb, A. K. Pattnaik, and S. E. Besra. “APOPTOSIS-INDUCING POTENTIAL OF LAWSONIA ALBA LAM. LEAVES ON HEPATOCELLULAR CARCINOMA (HEP-G2) CELLS ALONG WITH ITS ANTI-OXIDANT PROPERTY”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 8, no. 9, Sept. 2016, pp. 156-62, doi:10.22159/ijpps.2016.v8i9.12815.
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