In Vitro Cytotoxic Activity of Clinacanthus nutans Leaf Extracts Against HeLa Cells

Objective: This study was conducted to investigate the antiproliferative activity of extracts of Clinacanthus nutansleaves against human cervical cancer (HeLa) cells. Methods: C. nutans leaves were subjected to extraction using 80%methanol or water. The methanol extract was further extracted to obtain hexane, dichloromethane (DCM), and aqueousfractions. The antiproliferative activity of the extracts against HeLa cells was determined. The most cytotoxic extractwas furthered analyzed by apoptosis and cell cycle assays, and the phytochemical constituents were screened by gaschromatography-mass spectrometry (GC-MS). Results: All of the extracts were antiproliferative against HeLa cells, andthe DCM fraction had the lowest IC50 value of 70 μg/mL at 48 h. Microscopic studies showed that HeLa cells exposedto the DCM fraction exhibited marked morphological features of apoptosis. The flow cytometry study also confirmedthat the DCM fraction induced apoptosis in HeLa cells, with cell cycle arrest at the S phase. GC-MS analysis revealedthe presence of at least 28 compounds in the DCM fraction, most of which were fatty acids. Conclusion: The DCMfraction obtained using the extraction method described herein had a lower IC50 value than those reported in previousstudies that characterized the anticancer activity of C. nutans against HeLa cells.