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

Document Type : Research Articles


1 Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Penang, Malaysia.

2 Centre for Drug Research, Penang, Malaysia.

3 School of Chemistry, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.


Objective: This study was conducted to investigate the antiproliferative activity of extracts of Clinacanthus nutans
leaves 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 aqueous
fractions. The antiproliferative activity of the extracts against HeLa cells was determined. The most cytotoxic extract
was furthered analyzed by apoptosis and cell cycle assays, and the phytochemical constituents were screened by gas
chromatography-mass spectrometry (GC-MS). Results: All of the extracts were antiproliferative against HeLa cells, and
the DCM fraction had the lowest IC50 value of 70 μg/mL at 48 h. Microscopic studies showed that HeLa cells exposed
to the DCM fraction exhibited marked morphological features of apoptosis. The flow cytometry study also confirmed
that the DCM fraction induced apoptosis in HeLa cells, with cell cycle arrest at the S phase. GC-MS analysis revealed
the presence of at least 28 compounds in the DCM fraction, most of which were fatty acids. Conclusion: The DCM
fraction obtained using the extraction method described herein had a lower IC50 value than those reported in previous
studies that characterized the anticancer activity of C. nutans against HeLa cells.


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