Abstract: This study aimed to evaluate the cytotoxicity of a crude extract of Piper cubeba
against normal and breast cancer cell lines. To prepare the extract, P. cubeba seeds were
ground, soaked in methanol and dichloromethane and isolated by column chromatography.
Fractions were tested for cytotoxicity effects on normal fibroblast (L929), normal breast
(MCF-12A) and breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231).
The most effective fraction was selected for DNA fragmentation assay to detect apoptotic
activity. The results showed that the methanolic crude extract had a higher cytotoxic activity
against MDA-MB-468 and MCF-7 than a dichloromethane crude extract. Then, the methanolic
crude extract was separated into six fractions, designated A to F. Fraction C was highly
active against breast cancer cell lines with an IC50 value less than 4 μg/mL. Therefore,
Fraction C was further separated into seven fractions, CA to CG. The 1H-NMR profile
showed that Fraction CE was long chain hydrocarbons. Moreover, Fraction CE demonstrated
the highest activity against MCF-7 cells with an IC50 value of 2.69 ± 0.09 μg/mL and lower
cytotoxicity against normal fibroblast L929 cells with an IC50 value of 4.17 ± 0.77 μg/mL.
Finally, DNA fragmentation with a ladder pattern characteristic of apoptosis was observed
in MCF-7, MDA-MB-468, MDA-MB-231 and L929 cells, but not in MCF-12A cells.