Antiproliferative Effects of Artabotrys odoratissimus Fruit Extract and its Bioactive Fraction through Upregulation of p53/gamma H2AX Signals and G2/M Phase Arrest in MIA PaCa-2 Cells

Background: Artabotrys odoratissimus (Annonaceae) is a medicinal and ornamental plant widely cultivated in Southeast Asia for its famous ylang ylang essential oil. The fruits of this plant are used for health benefits, but very little is studied about the bioactive principles, their role in regulating oxidative stress and tumour progression. Objective: The study aimed to evaluate the antiproliferative effects of fruit extract of Artabotrys odoratissimus and its bioactive fraction using cell-based assays. Methods: The free radical scavenging and antiproliferative effects of Artabotrys odoratissimus fruit ethyl acetate (FEA) extract and its bioactive fraction were evaluated using cell viability assays, colony formation assay, double staining assay, reactive oxygen species (ROS) assay, comet assay, cell cycle analysis, and western blotting. Results: The extract showed phenolic content of 149.8 +/- 0.11 mu g/mg Gallic acid equivalents and flavonoid content of 214.47 +/- 4.18 mu g/mg Quercetin. FEA showed an IC50 value of 76.35 mu g/ml in the ABTS assay and an IC50 value of 134.3 +/- 7.8 mu g/ml on MIA PaCa-2 cells. The cells treated with 125 mu g/ml and 250 mu g/ml FEA showed increased apoptotic cells in Double staining assay, DNA damage during comet assay, enhanced ROS, and cell cycle arrest at G2M phase at 125 mu g/ml and 250 mu g/ml. The active fraction AF5 showed an IC50 value of 67 +/- 1.26 mu g/ml on MIA PaCa-2 cells during MTT assay, displayed potential antiproliferative effects, and showed a marked increase in the expression of gamma H2AX and p53. Conclusion: These results prove that the fruit extract and the bioactive fraction demonstrate oxidative stress-mediated DNA damage, leading to apoptosis in the MIA PaCa-2 cell line.