Cytotoxic Activity of Pumpkin (Cucurbita sp.) Seed Extract Liver Cancer Cell Line

Chin, Brian Yee Zheng (2023) Cytotoxic Activity of Pumpkin (Cucurbita sp.) Seed Extract Liver Cancer Cell Line. Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Liver cancer (Hepatocellular carcinoma, HCC) marked as the eighth most common type of cancer in Malaysia. Solutions are still being actively sought to lower the risk of getting liver cancer and plant-based extracts are most often the answer due to their natural bioactive compounds. Pumpkins are common plants in Malaysia and pumpkin seeds can be potentially associated with beneficial biological mechanisms due to the presence of bioactive compounds that show anthelmintic, antidiabetic, antidepressant, antioxidant, and anticancer properties. This project aims to obtain crude pumpkin seed extract by ethanolic soxhlet extraction method and investigate the cytotoxic activity of pumpkin seed extract to HepG2 human liver cancer cell line. Commercial organic pumpkin seeds (150g) were ground to powder form and were subjected to semi-continuous ethanolic extraction via soxhlet instrument and rotary evaporation. A total of 5.2% and 3.6% of crude pumpkin seed extract (PSE) were obtained from first and second extractions respectively. The first and second batches of PSE were subjected to evaluation of cytotoxic effect against HepG2 human liver cancer cell line via MTT assay, with treatment concentration range of (3.125 - 100 μg/mL) and (25 - 200 μg/mL) respectively, repeated for both PSE obtained. Based on the triplicate results and extrapolation of non-linear regression curve with the function of log (inhibitor) vs. normalized responses, the treatment concentration range of (3.125 - 100 μg/mL) at 48 hours showed a strong cytotoxic effect, which inhibits 50% of the HepG2 cells population at the concentration (IC50) of 30.60 μg/ml, evaluated at an R squared value of 0.5508 via graphpad prism software. However, statistical analysis done by one-way ANOVA (Tukey’s test) showed that the HepG2 cell viability does not differ significantly (p > 0.05) against both ranges of crude pumpkin seed extract concentrations for 24 and 48 hours. Multiple comparison data also showed no significant differences between different PSE concentrations within the same range. In conclusion, the IC50 value is not representative of the cytotoxic effect of PSE to HepG2 liver cancer cell line and the crude ethanolic pumpkin seed extract with concentration range of 3.125 - 100 and 25 - 200 μg/mL are not effective in terms of cytotoxicity induction to HepG2 human liver cancer cell. Further studies using solid-phase extraction methods in the purification of the main bioactive compounds - beta carotene from crude pumpkin seed extract are needed to validate the potential anticancer properties of pumpkin seed extract to various human cancer cell lines