Piper cubeba extract on triple negative breast cancer



J Pharm Pharmacogn Res 10(1): 39-51, 2022.

Original Article

Anticancer activity of Piper cubeba L. extract on triple negative breast cancer MDA-MB-23

[Actividad anticancerígena del extracto de Piper cubeba L. en el cáncer de mama triple negativo MDA-MB-231]

Saowanee Maungchanburi1, Thidarath Rattanaburee1, Yaowapa Sukpondma2, Aman Tedasen3, Varomyalin Tipmanee1, Potchanapond Graidist1*

1Department of Biomedical Sciences and Biomedical Engineering, Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand.

2Division of Physical Science and Center of Excellence for Innovation in Chemistry, Faculty of Sciences, Prince of Songkla University, Songkhla, Thailand.

3School of Allied Health Sciences, Walailak University, Thasala District, Nakhonsithammarat, Thailand.

*E-mail: gpotchan@medicine.psu.ac.th

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.

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Resumen

Contexto: Piper cubeba L. (familia Piperaceae) es utilizado tradicionalmente para el tratamiento de muchas enfermedades, incluido el cáncer.

Objetivos: Determinar la actividad anticancerígena de extractos crudos de P. cubeba sobre células cancerosas e identificar los componentes químicos de la fracción activa.

Métodos: Se extrajeron semillas de P. cubeba con diclorometano seguido de metanol y se purificaron mediante cromatografía en columna. Se examinó la citotoxicidad de las fracciones frente al cáncer de mama triple negativo (MDA-MB-468) usando un ensayo MTT. Luego, se evaluó la citotoxicidad de las fracciones activas frente a células de cáncer de mama (MCF-7 y MDA-MB-231), cáncer de colon (HT-29), colangiocarcinoma (KKU-M213) y fibroblasto normal (L929). Se investigó el contenido total de fenólicos, taninos y flavonoides de la fracción activa y se analizó la composición química mediante GC-MS. Se aplicó citometría de flujo para determinar el ciclo celular, la apoptosis y la evidencia de activación de caspasa.

Resultados: Las fracciones DE14 y DE15 mostraron actividad citotóxica contra MDA-MB-468. La fracción DE15 exhibió la citotoxicidad más fuerte contra MDA-MB-231, KKU-M213, HT-29 y L929. La composición de la fracción DE15, extraída con diclorometano, fue de ácidos grasos apolares y ésteres de ácidos grasos, seguidos de propilenglicol e hidrocarburos. Sin embargo, los compuestos fenólicos fueron detectados, lo que llevó a su propiedad antioxidante medible. Este extracto no provocó la detención del ciclo celular en MDA-MB-231. Sin embargo, la fracción DE15 aumentó significativamente las células apoptóticas a las 48 y 72 h, e indujo significativamente la actividad de caspasas múltiples en MDA-MB-231 de una manera dependiente del tiempo.

Conclusiones: La fracción DE15 exhibió efecto de citotoxicidad e indujo apoptosis en cáncer de mama triple negativo y mostró menos toxicidad en células de fibroblastos normales.

Palabras Clave: anticáncer; apoptosis; cáncer de mama triple negativo; Piper cubeba.

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Citation Format: Maungchanburi S, Rattanaburee T, Sukpondma Y, Tedasen A, Tipmanee V, Graidist P (2021) Anticancer activity of Piper cubeba L. extract on triple negative breast cancer MDA-MB-231. J Pharm Pharmacogn Res 10(1): 39–51.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)