ANTICANCER ACTIVITY, APOPTOTIC AND NECROSIS EFFECTS OF CHLOROFORM FRACTION AND SUBFRACTIONS OF SUNGKAI LEAF ON HT-29 AND ADENOCARCINOMA PRIMARY pT3N1cM1 COLON CANCER CELLS
Sungkai ( Peronema canescensJack) belongs to the Verbenaceaefamily has been used in a folk medicine by South Sumatran Community and Dayak Tribes in Borneo Island. Investigation on the chemical composition of Sungkai leaves revealed potential components for anticancer activities, however there is limited information on the anticancer activity of this plant. Preliminary screening showed that the chloroform fraction exhibited potential anticancer activity. The current study aims to investigate the anticancer activity of chloroform fraction and subsfractions, as well as the apoptotic and necrosis effects on the HT-29 and adenocarcinoma primary pT3N1cM1 colon cancer cells.
Th cytotoxic assay was carried out by MTT method. The antiproliferation, apoptotic, and necrotic activities of the samples were observed using flow cytometer. Metabolites identification of the active subfractions was carried out by LC-MS/MS analysis. The in silicoassay of metabolites in the active samples was carried out to predict drug-likeness (pkCSM online) and pharmacokinetic of the molecules.
The results of the cytotoxic assay showed that SFA3 has higher cytotoxicity than the other subfractions, but lower than the standard drug 5-Fu. Subfraction SFA3 gave selectivity index (SI) values of 29.22 and 865.00 in the HT-29 and primary adenocarcinoma pT3N1cM1cells, respectively compared to 5-FU with SI values of 5.00 and 10.00. The data suggested that SFA3 is more selective compared to 5-Fu in the HT-29 and primary adenocarcinoma pT3N1cM1cells.
Twenty compounds were identified in the SFA3 based on the LC-MD/MS analysis, consisting of terpenoids, steroids, phenylpropanoids, and xanthones. The in silicostudy on the compounds identified in SFA3 indicated that eighteen compounds has good drug-likeness properties, four compounds has good pharmacokinetic properties. In addition, five compounds of SFA3 (SN3, SN7, SN14, SN16 and SN18) showed better cytotoxic activity with a lower rerank score (RS) than the 5-Fu positive control. Besides that, the lethal dose value (LD 50) was obtained belonging to class 4 and 5 (low toxicity), no mutagenic effect, non-hepatotoxic and non-sensitizing to the skin, good ADE value, and no toxic effect on the target organ-receptor.
Key Words: Sungkai, HT-29 cells, pT3N1cM primary adenocarcinoma cells, cytotoxic, apoptosis, necrosis, in silico.