INVESTIGATION OF ANTICANCER ACTION OF ILLICIUM VERUM AGAINST HUMAN BREAST CANCER CELLS

Abstract

Cancer is one of the leading cause of mortality with breast cancer as the most crucial cause of death in females around the world including Pakistan. Though, nowadays different advanced treatment options for breast cancer are available but most of them are associated with severe side and toxic effects as well as high cost of the cancer treatment including chemotherapeutics is an important issue, particularly in developing countries like Pakistan. Plants are being used to discover medicines such as anticancer drugs from natural sources to cure and treat diseases. Illicium verum Hook. F (I.verum), belongs to Illiciacea family, is included in traditional medicine used for many types of ailments. Present study investigated the anticancer action and mechanism of dried fruit methanol extract of I. verum, against human breast cancer cells (MCF-7 and MDA-MB 231). To evaluate the anticancer action, MTT assay was performed to determine effect of I. verum extract on cell viability, cell proliferation and DAPI staining was used for the analysis of apoptosis in MCF-7 and MDA-MB-231 cells. To determine the mechanism of its anticancer action; against the specific reactive oxygen species including superoxide radical, hydrogen peroxide and hydroxyl radical scavenging activity assays were also conducted. In cell viability assay, different doses of I. verum methanol extract were used to treat the MCF-7 (0.25, 0.5, 1, 3, 6, 12, 25 and 50µg/ml) and MDA-MB-231(0.125, 0.25, 0.5, 1, 3, 6, 12 and 25µg/ml) cells. Both type of cells showed significant (p-value <0.01) decrease in cell viability in the presence of all tested doses, except for the dose of 0.25µg/ml of MCF-7 cells. In case of cell proliferation assay, IC50 doses (5.5µg/ml and 2.8µg/ml are for MCF-7 and MDA-MB-231, respectively) showed significant (p-value <0.01) decrease in cell proliferation and induction of apoptosis in both MCF-7 and MDA-MB-231 cells. However, the I. verum extract was found to be more effective to decrease the cell viability, proliferation and led to apoptosis against MDA-MB-231 cells. The key reactive oxygen species pertaining to cancer induction and proliferation, in vitro study including superoxide radical, hydrogen peroxide and hydroxyl radical scavenging assays’ data also showed that I. verum methanol extract showed significant (p-value <0.01) increase in percentage of superoxide radical, hydrogen peroxide and hydroxyl radical scavenging activity. I. verum extract was found to be most effective to scavenge hydrogen peroxide. ix In conclusion, I. verum methanol extract showed excellent anticancer action against both MCF-7 and MDA-MB-231 cells, however; the extract showed very promising anticancer action against MDA-MB-231 carcinoma cells. The I. verum methanol extract exerted its anticancer action via cytotoxicity, reduction in cancer cells growth, induction of apoptosis and superoxide radical, hydrogen peroxide and hydroxyl radical scavenging activity against these breast cancer cells. Therefore, we suggested that I. verum extract has great potential to discover effective chemotherapeutic agent(s) for breast cancer, particularly for triple native breast cancer. However, further studies are required to explore the active compound(s) of I. verum extract, as it is reported to contain many medicinally valuable compounds, which have potential like polyphenols, which may be responsible for its anticancer action.