Cytotoxicity of Clinacanthus nutans and Mechanism of Action of Its Active Fraction towards Human Cervical Cancer Cell Line, HeLA / Jurnal Sains Kesihatan Malaysia

ABSTRACT

@#Traditionally, Clinacanthus nutans (CN) or locally named as ‘Belalai Gajah’ is one of the herbal plant claimed to beable to treat cancer. The aimd of this study are to extract, isolate and characterize the active anticancer compoundfrom CN and to determine the mode of cell death induced by the compound. Bioassay guided fractionation was done onthe CN extract by using column chromatography. The cytotoxicity activities of these fractions toward HeLA cells wereexamined by MTT assay. The nuclear morphology was examined by Hoechst 33258 staining and the cell cycle arrestwas evaluated by propium iodide staining using flow cytometry. The presence of active compound in the chosen fractionwas determined by Liquid Chromatography Mass Spectrometry (LCMS). Out of 16 fractions collected, Fraction 11(F11)showed the lowest IC50 value with 27 ± 2.6 µg/mL. The value of IC50 for F11 towards normal cell, NIH 3T3 cell and L929cell, were 70 ± 4.0 µg/mL and 45 ± 1.5 µg/mL respectively. These values were higher than tamoxifen, therefore indicatingthat tamoxifen is more toxic towards normal cells compared to F11. Nuclear morphology of HeLA cell displayed DNAfragmentation, nuclear condensation and formation of apoptotic bodies upon treatment with F11 for 24 hours. The cellcycle distribution of HeLA cell treated with F11 was arrested at G1 phase. The active compound identified to potentiallypossess the anticancer property is 19-Oxo-all-trans-retinoic acid. In conclusion, 19-Oxo-all-trans-retinoic acids fromF11 of the CN extract, is a potential anticancer agent for cervical cancer.