Exploring the apoptosis-inducing potential of Cannabis sativa and cannabidiol in pancreatic cancer

Pancreatic cancer is deemed one of the most aggressive types of cancer, with a high mortality rate and poor prognosis. Pancreatic cancer is often diagnosed in advanced stages of which the disease is too aggressive to manage, rendering therapeutic interventions ineffective. Cannabis sativa (C. sativa) a medicinal plant popularly known for its psycho-altering characteristics is recently being explored for its anti-cancer properties. However, knowledge on its impact on pancreatic cancer is still limited. This study was aimed at exploring the apoptosis-inducing potential of C. sativa and cannabidiol (CBD) in pancreatic cancer cells. Crude C. sativa extracts were obtained by dissolving dried C. sativa leaves in absolute ethanol, CBD was isolated from the crude extract using column chromatography. The cytotoxic effects of the crude extract and CBD on pancreatic cancer cells (Mia-PaCa2) and normal lung cells (MRC-5) were determined using AlamarBlue reagent. Apoptosis was investigated by analysing morphological changes using light microscopy, Annexin V and Hoechst staining. ATP and caspase-3 and − 7 levels were also determined. DNA fragmentation was analysed using agarose gel electrophoresis. The expression of apoptosis related genes was assessed using quantitative polymerase chain reaction (qPCR) following CBD treatment. The results show that both the crude C. sativa and CBD induced substantial cytotoxic effects on Mia-PaCa2 cells but not MRC-5. These treatments caused significant morphological changes on Mia-PaCa2 cells causing the cells to detach, leading to cell death. The cells also stained positive with Annexin V, indicating an induction of apoptosis. Hoescht staining also confirmed nuclear condensation, another indication of apoptosis induction. Crude C. sativa and CBD led to reduced level of ATP as well as caspase-3 and − 7, with the effects more pronounced with the crude extract. DNA fragmentation was also more distinct in cells treated with the crude extract, validating the notion that both crude C. sativa and CBD induced apoptosis in Mia-PaCa2. This was further supported by the upregulation of BAX, BAK-1, and p53 and the downregulation of BCL-2 a indication of the intrinsic apoptotic pathway activation. In conclusion, both crude and CBD have apoptosis-inducing potential in pancreatic cancer cells.