Anticancer activity of Piper cubeba L. extract on triple negative breast cancer MDA-MB-231.

Abstract

Context: Piper cubeba L. (family Piperaceae) is used traditionally for the treatment of many diseases, including cancer. Aims: To determine the anticancer activity of P. cubeba crude extracts on cancer cells and identify chemical constituents of the active fraction. Methods: Seeds of P. cubeba were sequentially extracted with dichloromethane followed by methanol and purified using column chromatography. Fractions were screened the cytotoxicity against triple negative breast cancer (MDA-MB-468) using MTT assay. Then, active fractions were evaluated cytotoxicity against breast cancer (MCF-7 and MDA-MB-231), colon cancer (HT-29), cholangiocarcinoma (KKU-M213) and normal fibroblast (L929) cells. Total phenolic, tannin, and flavonoid contents of active fraction were investigated, and the chemical composition was analyzed using GC-MS. Flow cytometry was applied to determine the cell cycle, apoptosis, and evidence of caspase activation. Results: Fraction DE14 and DE15 showed cytotoxic activity against MDA-MB-468. Fraction DE15 exhibited the most potent cytotoxicity against MDA-MB-231, KKU-M213, HT-29 and L929. As the fraction DE15 was of dichloromethane extraction, major contents were nonpolar fatty acids and fatty acid esters, followed by propylene glycol and hydrocarbons. The phenolic compounds were however traceable, leading to their measurable antioxidant property. This extract did not cause cell cycle arrest on MDA-MB-231. However, fraction DE15 significantly increased apoptotic cells at 48 and 72 h, and significantly induced multi-caspases activity on MDA-MB-231 in a time-dependent manner. Conclusions: Fraction DE15 exhibited cytotoxicity effect and induced apoptosis on triple negative breast cancer and showed less toxicity on normal fibroblast cells.

Keywords

• piper cubeba
• anticancer
• apoptosis
• triple negative breast cancer