Abstract

Nineteen blood samples of clinically sickle cell patients were collected from, Ibn Al-Baladi hospital, in Iraq. Besides, blood samples were taken from 10 apparently healthy individuals (without Hemoglobinopathy disorders (as a control group. The apoptotic response shown by sickle cells was evaluated using three methods, (DNA fragmentation assay, Comet assay and Annexin V-FITC (Fluorescein isothiocyanate) apoptosis). The results obtained by DNA fragmentation assay reveal in severe DNA damage of the sickle patients according to the molecular weight of the ladder. The smear shape pattern on gel electrophoresis indicating double strand breakage of the DNA was detected of 39.5% in studied sickle cell samples. . Compared to the control and MW of ladder, the sickle cell patients had higher prevalence of DNA double-strand breaks in their leukocytes. The other experiment of apoptosis study involved the utilization of Annexin-V-FITC kit to monitor apoptotic and necrotic induction in sickle cells. The results showed high apoptotic percentages in sickle cell patients (34.5%) compared to healthy people. Comet assay result was demonstrated as comet index which represent a mean of the scored which calculate from the ratio of cell diameters (L/W) of total cells in each sample. These results revealed that lymphocytes cells exhibited an increase apoptosis percentage (37.5.2%) compare with percentage of control (8%). This revealed that there was DNA damage in sickle cell patients.

Keywords:

Apoptosis, comet assay, DNA damage, DNA fragmentation, flow cytometry, polymorphonuclear leukocytes, sickle cell disease,

References

Competing interests

The authors have no competing interests.

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The authors have no competing interests.