PHYTOCHEMICAL PROFILING, ANTICANCER AND APOPTOTIC ACTIVITY OF GRAVIOLA (ANNONA MURICATA) FRUIT EXTRACT AGAINST HUMAN HEPATOCELLULAR CARCINOMA (HepG-2) CELLS

Abstract

To examine the phyto-constituents present in A. muricata fruit pulp, and to assess the anticancer potential followed by apoptotic studies on HepG-2 cell line. Aqueous, Chloroform, Ethyl acetate, Hexane and Methanol extracts of dragon fruit pulp were analysed by qualitative phytochemical profiling and anti-proliferation activity. Further, the apoptotic study of methanol extract treated HepG-2 cells by nuclear staining using AO, DNA fragmentation, ATR and GC-MS spectral analysis for methanol extract was carried out. The preliminary phytochemical screening showed the presence of acids, alkaloids, carbohydrate, flavonoids, phenols, protein, saponins, tannins, terpenoids and triterpenoids in aqueous extract. Likewise, acids, alkaloids, carbohydrates, proteins, saponins, steroids and tannins were present in chloroform extract. On the other hand acids, carbohydrate, flavonoids, phenols, protein and tannins were present in ethyl acetate extract. Similarly, acids, alkaloids, carbohydrate, flavonoids, protein, saponins and tannins were present in hexane extract, and acids, alkaloids, carbohydrate, coumarins, flavonoids, glycosides, phenols, protein, saponins, steroids, tannins, terpenoids and triterpenoids were present in methanol extract. Dose dependant anti-proliferative assay revealed that, The IC50 value was 62.699 μǥ/ml in aqueous extract, 63.710 μǥ/ml in chloroform extract, 20.617 μǥ/ml in ethyl acetate extract, 44.553 μǥ/ml in hexane extract and 13.104 μǥ/ml in methanol extract at 24 h incubation. Methanol fruit extract of A. muricata showed intense fragments of nucleus as signs of apoptosis by AO staining. Apoptosis can be visualized as a ladder pattern of 100-200 bp due to DNA cleavage by the activation of a nuclear endonuclease by standard agarose gel electrophoresis. The results of ATR revealed the presence of alkyl halides, amines, carboxylic acids, phenols and alcohols. GC-MS results revealed 11 volatile compounds present in methanol extract. Methanol extracts of A. muricata has confirmed hopeful anticancer activity against human liver cancer (HepG-2) cells by in vitro method.