GC-MS ANALYSIS AND IN-VITRO CYTOTOXIC STUDIES OF BIXA ORELLANA SEED EXTRACT AGAINST CANCER CELL LINE
Keywords:
Bixa orellana, GC-MS, Geranylgeraniol, Comet assay, Annexin VPI staining, AOEB stainingAbstract
Objective: The present study was carried out to analyze the bioactive component of petroleum ether (PE) extract of Bixa orellana seed and its anticancer potential against B16F-10 melanoma cell line.
Methods: The presence of a bioactive component of PE extract was analyzed by GC-MS method. Cell viability was measured using MTT assay. Apoptotic inductive effect of the extract was evaluated by AO/EB staining; DAPI is staining and Annexin V/PI staining. DNA fragmentation analysis of the control and treated cell was carried out by agarose gel electrophoresis and comet assay. In vitro prevention of cell proliferation and migration was further estimated by colony forming assay and wound healing assay respectively.
Results: The cytotoxicity of the PE extract against B16F10 melanoma cell was dose and time dependent. The IC50 values for the B16F-10 melanoma cell were 145.98±7.49 µg/ml and 121.60±6.20µg/ml for 24h and 48h respectively. DAPI staining showed DNA fragmentation which was further confirmed by ladder-like appearance of DNA by gel electrophoresis and comet assay. PE extract has prevented the cell proliferation and migration of the treated cancer cell. GC-MS analysis shows the presence of three anticancer compound geranylgeraniol, squalene and beta-sitosterol.
Conclusion: The present study established that PE extract of Bixa orellana seed possesses potent anticancer and apoptotic inductive potential which can further be explored in-vivo model for possible cytotoxic activity.
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