An ethanol extract of Hawaiian turmeric: Extensive in vitro anticancer activity against human colon cancer cells

Abstract

Context Turmeric (Curcuma longa) is a food spice and colorant reported to be beneficial for human health. Curcumin (diferuloylmethane) is the major ingredient in turmeric, and existing data suggest that the spice, in combination with chemotherapy, provides a superior strategy for treatment of gastrointestinal cancer. However, despite its significant effects, curcumin suffers from poor bioavailability, due to poor absorption in the body. Objective The research team intended to evaluate a liquid extract of turmeric roots (TEx) that the team had formulated for its in vitro, anticancer activity against several human, colorectal cancer cell lines. Design The research team performed in vitro studies evaluating the anticancer efficacy via short and long-term assays and also evaluated invasion using Matrigel (Corning Life Sciences, Tewksbury, MA, USA). Further, in vitro anticancer activity of TEx was tested against 3-D cultures of HCT166 spheroids, which were subsequently analyzed by flow cytometry. Setting ADNA, Inc, Columbus, OH, USA; Foundation for Biomedical Research of the Academy of Athens, Athens, Greece; and Laboratory of Pharmacology, Faculty of Medicine, University of Thessaly, Larissa, Greece. Intervention The study used 4 human cell lines of colorectal cancer—HT29, HCT15, DLD1, and HCT116— and 2 breast cancer cell lines—SW480 and MDA-MB231. For a short-term assay, the extract was dissolved into culture mediums of HT29, HCT15, DLD1, HCT116, and SW480 at four 10-fold dilutions (100 to 0.1 μg/mL). For a long-term assay, TEx was added to the cultures of the same cell lines at 3 dilutions—20, 10, and 5 μg/mL. For an invasion assay, 100 μL per well of Matrigel was added and allowed to polymerize prior seeding of the MDA-MB231 cells. For cultures treated with the TEx, the TEx was mixed with the cell suspension prior to the seeding step. For the spheroid testing, the TEx was added to HCT116 cells either at the beginning of an experiment (ie, before the addition of the cancer cells), which was a chemopreventive approach, or 48 h later, on the addition of cells to the wells to allow the generation of spheroids, which was a chemotherapeutic approach. Outcome Measures The in vitro activities of TEx were evaluated using a 48-h-incubation, short-term assay and a 2-wk, long-term (clonogenic) assay. To analyze the antiinvasive activity of the extract, images for the Matrigel invasion assay were taken with a camera at the 24-h time point. The in vitro, anticancer activity of TEx was also tested against 3-D cultures of HCT116 spheroids that were subsequently analyzed using flow cytometry. Results TEx had potently inhibited the growth of all human colon cancer cell lines tested in a dose- and timedependent manner. TEx inhibited the formation of HCT116 spheroids when the cells were incubated with the extract. The extract also disrupted the formation of tubules formed by MDA-MB231 cells grown on Matrigel at concentrations that did not affect the overall viability of the cells, indicating a potent anti-invasive activity. Conclusions These data suggest a potential therapeutic activity for TEx against human colon cancer, most likely due to the enhanced bioavailability of the turmeric. © 2015, InnoVision Communications. All right rserved.