The IN VITRO STUDY OF APOPTOSIS INDUCTION BY L-ASPARAGINASE OF ASPERGILLUS FUMIGATUS (ASPF-6) ON HUMAN LEUKEMIA CELL LINES
On Human Leukemia cell lines,HL-60
Keywords:L-asparaginase,, A fumigatus,, 3-(4,5-dimethyl thiazol-2-)-2,5-diphenyl tetrazolium bromide assay,, Annexin VPI and JC-1 Flurocytometry
Objective: L-Asparaginase is a relatively widespread enzyme used in the lymphoblastic leukemia chemotherapy. In the present investigation, we report new potential fungal L-asparaginase producer Aspergillus fumigatus from Kadalundi mangrove forest, Kerala. The aim of the present investigation was to evaluate the concentration-dependent cytotoxicity and apoptosis-inducing the effect of extracted enzyme and its ability to induce apoptosis against human leukemia cell lines-HL-60.
Methods: The fungal strain was isolated by serial dilution method from rhizosphere soil of mangrove forest. The isolated enzyme purified by 80% ammonium sulfate precipitation, dialysis, and ion exchange chromatography. To determine cytotoxicity and level of apoptosis, three different tests were performed: (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, JC-1 flow cytometry, and Annexin V/propidium iodide (PI) flow cytometric DNA fragmentation. The test compound efficiency compared with positive drug doxorubicin against HL-60 cell lines.
Results: The test compound exhibited a concentration-dependent cytotoxic effect on proliferation of HL-60 cell lines (IC-50 12.39 U) with a different level of apoptosis induction (LR-11.3%).
Conclusion: A. fumigatus derived L-asparaginase may be clinically useful and results in better utilization for limited fungal-derived drug resources and improve financial feasibility as a therapeutic option for human leukemia.
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