In Vitro Antitumor Properties of an Isolate from Leaves of Cassia alata L

  • Published : 2013.05.30


Leaf extracts of Cassia alata L (akapulko), traditionally used for treatment of a variety of diseases, were evaluated for their potential antitumor properties in vitro. MTT assays were used to examine the cytotoxic effects of crude extracts on five human cancer cell lines, namely MCF-7, derived from a breast carcinoma, SK-BR-3, another breast carcinoma, T24 a bladder carcinoma, Col 2, a colorectal carcinoma, and A549, a nonsmall cell lung adenocarcinoma. Hexane extracts showed remarkable cytotoxicity against MCF-7, T24, and Col 2 in a dose-dependent manner. This observation was confirmed by morphological investigation using light microscopy. Further bioassay-directed fractionation of the cytotoxic extract led to the isolation of a TLC-pure isolate labeled as f6l. Isolate f6l was further evaluated using MTT assay and morphological and biochemical investigations, which likewise showed selectivity to MCF-7, T24, and Col 2 cells with $IC_{50}$ values of 16, 17, and 17 ${\mu}g/ml$, respectively. Isolate f6l, however, showed no cytotoxicity towards the non-cancer Chinese hamster ovarian cell line (CHO-AA8). Cytochemical investigation using DAPI staining and biochemical investigation using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-a method used to detect DNA fragmentation-together with caspase assay, demonstrated apoptotic cell death. Spectral characterization of isolate f6l revealed that it contained polyunsaturated fatty acid esters. Considering the cytotoxicity profile and its mode of action, f6l might represent a new promising compound with potential for development as an anticancer drug with low or no toxicity to non-cancer cells used in this study.


  1. Agarwal C, Singh RP, Agarwal R (2002). Grape seed extract induces apoptotic death of human prostrate carcinoma DU 145 cells via caspases activation accompanied by dissipation of mitochondrial membrane potential and cytochrome C release. Carcinogenesis, 23, 1869-76.
  2. Alshatwi AA, Shafi G, Hasan TN, et al (2011). Apoptosismediated inhibition of human breast cancer cell proliferation by lemon citrus extract. Asian Pac J Cancer Prev, 12, 1555-9.
  3. Chiu LC, Wan JM, Ooi VE (2000). Induction of apoptosis by dietary polyunsaturated fatty acids in human leukemic cells is not associated with DNA fragmentation. Int J Oncol, 17, 789-96.
  4. Hanada H, Morikawa K, Hirota K, et al (2011). Induction of apoptosis and lipogenesis in human preadipocyte cell line by n−3 PUFAs (abst.). Cell Biol International, 35, 51-9
  5. Huai J, Jockel L, Schrader K, Borner C (2010). Role of caspases and non-caspase proteases in cell death. F1000 Biol Rep, 2.
  6. Ibrahim D, Osman H (1995). Antimicrobial activity of Cassia alata L from Malaysia. J Ethnopharmacol, 45, 151-6.
  7. Jacinto SD, Olarte EI, Galvez M, et al (2005). Leaf extracts from Cassia alata L ("Akapulko") induces quinone reductase and competes for estrogen receptor binding indicating cancer chemopreventive property. Philippine Agricultural Scientist, 88, 473-6.
  8. Janicke RU (2009). MCF-7 breast carcinoma cells do not express caspase-3. Breast Cancer Res Treat, 117, 219-21.
  9. Matic I Z, Aljancic I, Zizak Z, et al (2013). In vitro antitumor actions of extracts from endemic plant Helichrysum zivojinii. BMC Complementary and Alternative Medicine, 13, 36.
  10. Mosmann T (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65, 55-63.
  11. Pereira, WO, Amarante-Mendes G P (2011). Apoptosis: a programme of cell death or cell disposal? Scand J Immunol, 73, 2513.
  12. Pessoa C, Silveira ER, Lemos TLG, et al (2000). Antiproliferative effects of compounds derived from plants of northern Brazil. Phytother Res, 14, 187-91.<187::AID-PTR572>3.0.CO;2-I
  13. Phillips, C (2011). Study raises concerns about using cancer cell lines to test drug resistance. NCI Cancer Bull, 8, 5.
  14. Roberts R (2000). Apoptosis in toxicology. Taylor and Francis NY, 10, 24-8.
  15. Samejima K, Kottke T, Enari M, et al (1998). Transition from caspase-dependent to caspase-independent mechanisms at the onset of apoptotic execution. J Cell Biol, 143, 225-39.
  16. Serini S, Piccioni E, Merendino N, et al (2009). Dietary polyunsaturated fatty acids as inducers of apoptosis: implications for cancer. Apoptosis, 14, 135-52
  17. Siddiqui AR, Zerouga M, Wu M, et al (2005). Anticancer properties of propofol-docosahexaenoate and propofol - eicosapentaenoate on breast cancer cells. Breast Canc Res, 7, 645-54.
  18. Undurti ND, Madhavi N (2011). Effect of polyunsaturated fatty acids on drug-sensitive and resistant tumor cells in vitro, Lipids Health Dis, 10, 159.
  19. Villasenor IM, Canlas AP, Pascua MPI, et al (2002). Bioactivity studies on Cassia alata Linn Leaf extracts. Phytother Res, 16, 93-6.

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