Aqueous Fraction of <em>Nephelium ramboutan-ake</em> Rind Induces Mitochondrial-Mediated Apoptosis in HT-29 Human Colorectal Adenocarcinoma Cells

• Main Authors: Muhamad Noor Alfarizal Kamarudin, Habsah Abdul Kadir, Bey Hing Goh, Chim Kei Chan
• Format: Article
• Language: English
• Published: MDPI AG 2012-05-01
• Series: Molecules
• Subjects: <em>Nephelium ramboutan-ake</em> pulasan; cytotoxicity; apoptosis
• Online Access: http://www.mdpi.com/1420-3049/17/6/6633

The aim of this study was to investigate the cytotoxic and apoptotic effects of <em>Nephelium ramboutan-ake</em> (pulasan) rind in selected human cancer cell lines. The crude ethanol extract and fractions (ethyl acetate and aqueous) of <em>N. ramboutan-ake</em> inhibited the growth of HT-29, HCT-116, MDA-MB-231, Ca Ski cells according to MTT assays. The <em>N. ramboutan-ake</em> aqueous fraction (NRAF) was found to exert the greatest cytotoxic effect against HT-29 in a dose-dependent manner. Evidence of apoptotic cell death was revealed by features such as chromatin condensation, nuclear fragmentation and apoptotic body formation. The result from a TUNEL assay strongly suggested that NRAF brings about DNA fragmentation in HT-29 cells. Phosphatidylserine (PS) externalization on the outer leaflet of plasma membranes was detected with annexin V-FITC/PI binding, confirming the early stage of apoptosis. The mitochondrial permeability transition is an important step in the induction of cellular apoptosis, and the results clearly suggested that NRAF led to collapse of mitochondrial transmembrane potential in HT-29 cells. This attenuation of mitochondrial membrane potential (Δ<em>ψ</em>m) was accompanied by increased production of ROS and depletion of GSH, an increase of Bax protein expression, and induced-activation of caspase-3/7 and caspase-9. These combined results suggest that NRAF induces mitochondrial-mediated apoptosis.