Immunomodulatory and anticancer potential of a traditional medicine product from a traditional healer against MCF-7 breast cancer cells.

Abstract

The incidence of breast cancer in South Africa is expected to rise dramatically in the forthcoming decade. In 2018, approximately 2 088 849 new cases and 626 679 cases of deaths from breast cancer alone were reported worldwide. Among African countries, the incidence of cancer is higher in southern Africa with an estimate standard rate of 235.9 per 1000, 0000 people. Currently breast cancer is treated with either chemotherapy, surgery, radiotherapy or immunotherapy. All this conventional treatment options available for breast cancer have some limitations and drawbacks. Therefore, there is an urgent need for the development of novel breast cancer therapy that will potential kill tumour cells more effective and selective. The use of traditional medicine as an alternative source of healthcare is gaining an increasing interest in many countries including South Africa. This is due to their accessibly and perceived effectiveness. However, the majority of these traditional medicines have not been scientifically validated. Aim: The purpose of this study was to determine immunomodulatory and anti-cancer activity of an African traditional medicine product from a traditional healer (Product Nkadinde, a traditional medicine composed of four medicinal plants) on MCF-7 breast tumour cells. Methods: Normal peripheral blood mononuclear cells (PBMCs) were used initially to establish the IC50 dose of Product Nkabinde using the Celltiter-Glo™ ATP assay. Conditioned media from PBMCs was then prepared by treating a separate set of normal PBMCs using the IC50 dose and the positive control, camptothecin (1μM). For the anticancer assays, the Celltiter-Glo™ ATP assay was used to determine the cytotoxicity effects of conditioned media from PBMCs with Product Nkabinde against MCF-7 breast cancer cells. Antioxidant levels (changes in intracellular glutathione (GSH) levels) were measured using GSH-Glo™ Glutathione assay kit. The mechanism of cell death was determined using the Caspase 3/7 and DNA fragmentation assays. Immunomodulation studies was carried out using a multi analyte cytokine ELISA assay kit. Results: The results obtained from this study revealed that Product Nkabinde was cytotoxic to MCF-7 and an IC50 dose was established at 344 μg/ml in treatment PBMCs. Product Nkabinde was able to induce caspase 3/7 activity, decrease intracellular GSH levels and induce DNA fragmentation comparative to the positive control, thus showed potential to induce apoptosis in breast cancer cells. Inflammatory cytokines secretion studies indicated that Product Nkabinde did not significantly change the secretion of all 12 inflammatory cytokines analyzed in treated PBMCs. However, Product Nkabinde was shown to increase the secretion of IL 1β, IL 8 and GM-CSF in treated MCF-7 cells, although this was not significant. Conclusion: The results of this study showed that Product Nkabinde induced immunomodulatory effects through changes in cytokines secretion and direct anticancer effects through activation of caspase 3/7 enzymes, decreasing GSH levels and induction of DNA fragmentation in treated MCF-7 cells. This traditional medicine has the potential to be developed and optimized as an alternative anticancer medicine.