The APOPTOSIS EFFECT OF LUVUNGA SCANDENS AGAINST HUMAN SKIN CANCER A431 CELL LINE
SAMA NAZIYAH SHABAN
sama.nazih@uniq.edu.iq
Abstract:
Epidermal carcinoma cell line is one of the neoplastic cell that are characterized by abnormalities in cell differentiation and growth. Epidermal carcinoma is reported as one of the most common types of cancer with increasing numbers of occurrence. Luvunga scandens is one of the plants found in Malaysia, this plant is known to possess many bioactivities and general health effects, yet its anti-proliferative effect is generally under reported and need to be scientifically evaluated. The aim of this study is to evaluate the anti-proliferative and apoptotic effects of Luvunga scandens plant leaves against human epidermal carcinoma cell line. MTT assay was used to assess the cytotoxicity of the plant against human epidermal carcinoma cells in addition to the safety assessment for normal cell lines (HaCaT and HDF). Scratch assay was carried out to monitor the cell growth. The morphological changes of L. scandens treated epidermal carcinoma cells has been confirmed by SEM, and the apoptosis of the plant against epidermal carcinoma cells has been tested using caspase 3/7 assay, followed by cell cycle analysis done using a flow-cytometer on epidermal carcinoma cells treated with the IC50 does of L. scandens plant. Western blot was preformed to confirm the anti-carcinogenic effect of L. scandens against human epidermal carcinoma cells. The results of the tests done illustrated that the extract and compound possesses cytotoxic effect against epidermal carcinoma cells with IC50 readings; (methanol= 37.5 µg/mL, DCM= 38 µg/mL, hexane= 37.5 µg/mL and compound= 27.5 µM), and no cytotoxic activity in both HaCaT and HDF cells. The IC50 dose of L. scandens can restrict the growth of epidermal carcinoma cells more than Cisplatin (anti-cancer drug), the SEM results demonstrate that L. scandens treated cells showed an overall change in the cell shape, alteration of surface morphology, absence of microvilli and appearance of blebs. Caspase 3/7 assay results shows that L. scandens DCM extract produce the highest level of apoptosis against epidermal carcinoma cell. For cell cycle analysis, all the L. scandens treated epidermal carcinoma cells show high readings in the sub-G1 phase. For western blot the L. scandens extract and compound show high apoptosis effects against human epidermal carcinoma cells. This in vitro study has proved that L. scandens plant exhibit antiproliferative effects against human epidermal carcinoma cells, hence, it can be considered as a new promising potential anti-cancer therapy.
