Purpose: To evaluate the anti-cancer activity of Limonia acidissima Linn (Rutaceae) which has long been used traditionally for various infectious and malignant diseases.

Methods: The fruit extract of Limonia acidissima Linn was obtained by macerating 3 times with methanol and then concentrating it at reduced pressure. Bioassay of the extract was carried out to assess the anticancer activity. Breast cancer cell lines, SKBR3 and MDA MB435, were used for in-vitro cell proliferation, cell viability assay, and cell cycle analysis of the extract.

Results: Bioassay of the extract of Limonia acidissima Linn.  showed that a fraction (fraction 3) of the ethanol extract had anticancer activity against SKBR3 and MDA-MB435 human breast cancer cells. The effective dose (ED₅₀) of Limonia acidissima Linn. fraction 3 was 56.1 and 30.6 μg/ml for SKBR3 and MDA-MB435, respectively. After 48 h of exposure, this fraction (100 μg/ml) significantly reduced cell proliferation in both cancer cell lines. In MDA-MB435 cells, cell cycle analysis showed that fraction 3 induced the accumulation of cells in G2/M phase, but no significant change in cell cycle was detected in SKBR3 cells.

Conclusion: The results indicate that the extract fraction could induce cell cycle arrest. However, further investigation is needed to assess the molecular mechanisms of the anticancer activities of this plant.