Abstract

Cancer is an enormous burden of disease globally. Today, more people die from cancer than a combination of several diseases. And in females, breast and cervical malignancies remain the most common types. Currently, cervical and breast cancer are the most diagnosed gynecological cancer type amongst black females in the Southern Sahara while amongst males prostate cancer is on the upward trend. With many of them still dependent on medicinal plants as a form of therapy and the need to identify new therapeutic agents, we have identified a commonly used medicinal plant Tulbaghia violacea Harv. commonly known as Itswele lomlambo (Xhosa), wilde knoffel (Afrikaans) and Isihaqa (zulu) to evaluate its anticancer properties at a molecular biology level. In this study, we evaluated the molecular mechanism of T. violacea extracts in regulating cell death in various cancer cell lines. To achieve this, T. violacea was collected, dried before crushing into a fine ground powder. Three organic solvents namely, methanol, hexane, and butanol at 10 g per 100 mL were used as extraction solvents. Each cell line was treated with varying concentrations of the plant extract to identify the half-maximal inhibitory concentration (IC50). The IC 50 was later used to analyse if the extracts were inducing apoptosis using annexin V analysis. Furthermore, the molecular mechanisms by which apoptosis was induced was analysed by qPCR, western blots. All three extracts exhibited anticancer activity with the most cytotoxic being methanol extract. p53 expression was significantly increased in treated cells that correlated with increased caspase activity. The results point to possible activation of apoptosis following treatment with hexane extracts.

Highlights

  • Cancer is an enormous burden of disease globally

  • As expected, we can see that the different solvents used for extraction, induced differential secondary metabolite changes as demonstrated by increases or decreases in peak intensities and the appearance of new molecular ion peaks and suppression of other molecular ion peaks, which are present in the Base Peak Intensities (BPI)’s of other chromatograms

  • Apoptosis activation can either be intrinsic or extrinsic, so we evaluated the expression of Fas and caspase 8, which are mainly involved in the activation of the extrinsic pathway; we observed a significant increase in Fas in HeLa cells but a smaller increase in all other cell lines, while caspase 8 was steadily increased in all cell lines (Fig. 8G, H)

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Summary

Introduction

Cancer is an enormous burden of disease globally. Today, more people die from cancer than a combination of several diseases. We evaluated the molecular mechanism of T. violacea extracts in regulating cell death in various cancer cell lines. The results point to possible activation of apoptosis following treatment with hexane extracts. A successful anticancer drug should kill cancer cells without causing unnecessary damage to normal cells, and that can only be achieved by restoring the apoptosis machinery in cancer ­cells[4]. Studies have previously demonstrated the cytotoxic activities of extracts of T. violacea (Wild garlic from Southern Africa) in cancer c­ ells[5]. It is imperative that apoptotic inducers be screened from plants, either in the form of crude extracts or as compounds isolated from t­hem[8]. In this study, we evaluated the apoptotic induction potential of Tulbaghia violacea (TV) in various cancers. The extraction yield (%) was calculated as f­ollows[9]: Weight of the extract after evaporating solvent and freeze drying

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