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

  • VIPINA R Department of Microbiology, See Narayana Guru College, Coimbatore, Tamil Nadu, India.
  • VIJAYA CHITRA A Department of Microbiology, See Narayana Guru College, Coimbatore, Tamil Nadu, India.

Abstract

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.

Keywords: L-asparaginase,, A. fumigatus,, 3-(4,5-dimethyl thiazol-2-)-2,5-diphenyl tetrazolium bromide assay,, Annexin V/PI and JC-1 Flurocytometry.

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VIPINA R, and VIJAYA CHITRA A. “The IN VITRO STUDY OF APOPTOSIS INDUCTION BY L-ASPARAGINASE OF ASPERGILLUS FUMIGATUS (ASPF-6) ON HUMAN LEUKEMIA CELL LINES”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 7, May 2019, pp. 188-93, https://innovareacademics.in/journals/index.php/ajpcr/article/view/32158.
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