Cytotoxic Activity of Peronema canescens Jack Leaves on Human Cells: HT-29 and Primary Adenocarcinoma Colon Cancer

Abstract

Background: In Indonesia, this species was well known in Sumatera, Kalimantan, Java, and Sulawesi. Peronema canescens Jack (Sungkai) was traditionally used as an anti-flatulent, fever, toothache. Sungkai leaves contain many secondary metabolites with potential anticancer activity. The reported anticancer research was still limited to the cytotoxic activity of chloroform extract on the HT-29 colon cancer cell line. However, it was necessary to uncover the underlying mechanism. Aim: The purpose of this study was to investigate the mechanism (such as cell cycle inhibition, induces cells apoptosis, and necrosis) of subfraction chloroform (SF3) from P. canescens extract has anticancer activity on HT-29 cells and primary Adenocarcinoma (AdenoCa pT3N1cM1) colon cancer cells. Materials and Methods: The extraction by maceration method using methanol solvent, the fractionation process was using vacuum column chromatography (VCC) with polarity gradient eluent. The cytotoxicity of SF3 was measured by MTT assay. The cell cycle inhibition, apoptosis induction, and necrosis cells were evaluated with the Flow cytometry method. Results: Cytotoxicity value (IC50) against AdenoCa cells was 1.897 μg/ml. The inhibition activity of synthesis and mitosis phase in cell cycle demonstrated that the different concentrations of SF3 have inhibition activity on HT-29 (29.614 μg/ml) of 26.79% and 0.16%, AdenoCa cells (14.807 μg/ml) of 10.27% and 19.29%, respectively. For induced apoptosis activity on HT-29 (29.614 μg/ml) and AdenoCa cells (14.807 μg/ml) were 26.58% and 11.50%, successively. Whereas, necrosis activity on HT-29 (29.614 μg/ ml) and AdenoCa cells (14.807 μg/ml) were 0.02%, and 9.56%, respectively. Conclusion: The subfractions chloroform (SF3) of P. canescens extract has potential activity on HT-29 and Adenocarcinoma cells through cell cycle inhibition, induces apoptosis and necrosis cells. Key words: Apoptosis, Cell cycle, Colon cancer cells, Necrosis, Peronema canescens Jack