Morphological and molecular effects of phenolic extract from coconut kernel on human prostate cancer cell growth in vitro

Abstract

Coconut is an indispensable ingredient in the diet and traditional medicine of individuals belonging to the Indian subcontinent. Coconut is of high nutritional value owing to the presence of all essential dietary components, viz, saturated fatty acids, arginine rich proteins, fibre and minor components like vitamin E, phytosterols, polyphenols and flavonoids. The polyphenolic content present in coconut kernel is of particular interest due to their numerous reported beneficial effects such as reduction of oxidative stress, combating cancer and in modulating anti-inflammatory pathways. Therefore, in the present study the cytotoxic effect of the polyphenol rich fraction from coconut kernel (CKf) was evaluated in human prostate cancer (DU-145) cells. Individual components present in CKf was determined by LC-MS analysis. It showed that CKf contained several bioactive molecules which have potential anticancer activity viz, coumaric acid, myristin, chlorogenic acid and triterpenoid methyl esters. The cytotoxic effect of CKf at various concentrations (2.5–20 μg/ml) on DU-145 was assessed using MTT assay, AO/EB staining, mitochondrial superoxide/ROS production and changes in intracellular calcium levels, 24 hrs post treatment. Changes in the cell morphology and nucleus were observed using Scanning Electron Microscopy and Confocal microscopy. ROS and mitochondrial superoxide levels was evaluated using DCHF-DA and MitoSOX staining respectively. The impact of ROS on changes in cellular calcium levels was also studied using Fura-2-AM. LDH leakage from C K f treated and control cells were observed colorimetrically. Further, PCR analysis was done to detect changes in mitochondria associated apoptotic gene expression. It was also observed that C K f treatment increased the expression of pro-apoptotic genes - Bax, Bid, Bak and p53 in a dose-dependent manner. Based on the above results, it can be concluded that C K f may be used as a part of a dietary regime for controlling the progression of prostate cancer.