cute myeloid leukemia, with an annual incidence of about 3.5 per 100,000 people, is the most common form of acute leukemia in adulthood [4, 5]. In this heterogeneous cancer, in terms of phenotype and genotype, due to genetic changes in blood precursor cells, the natural growth and differentiation of these cells is altered and causes the accumulation of abnormal and immature myeloid cells in the bone marrow and peripheral blood [6]. 
Polymorphisms in a gene locus expressing proteins involved in the development and activation of myeloid cells may play a role in the immaturity of these cells. The ZAP-70 gene provides instructions for the production of the zeta-chain-dependent protein kinase, which is part of the signaling pathway for the evolution and activation of blood cells. rs104893674 (A/C)  is a single nucleotide polymorphism of the ZAP-70 gene, which due to its location inside the exon can directly increase or decrease the function of the gene and subsequently increase or decrease the susceptibility to disease.

This case-control study was performed on 94 patients with acute myeloid leukemia as a case group and 99 healthy individuals as a control group. All adults with acute myeloid leukemia diagnosed by a specialist, were included in the patient group. Also, people without cancer who were age- and sex-matched with cancer patients were included in the control group. Individuals with a history of any internal disease were excluded from the study. A total of 5 ml of peripheral blood was collected from the subjects. DNA was then extracted by “salting out” method and transferred to the freezer at -20° C. The quality of the extracted DNA was determined on 1% agarose gel and the rs104893674 (A/C) polymorphism was determined using Tetra-primer ARMS PCR technique. Data were analyzed using SPSS v. 23 and Chi-square statistical test.

The results of Tetra-primer ARMS PCR test showed 169 bp amplified fragments for CC genotype and 237 bp for AC genotype resulting from ZAP-70 gene amplification at position rs104893674 (A/C).
Since no AA genotype was found in the samples, no amplified product was shown on the gel. The results of chi-square test showed a significant relationship between the control group and the patient group in the frequency of both alleles A and C in the polymorphism rs104893674 position (P=0.000).



There was a significant relationship between the control group and the patient group in the frequency of CC and AC genotypes in the polymorphism rs104893674 position (P=0.000) (Table 1 & 2).


 

In acute leukemia, the bone marrow produces large amounts of immature white blood cells, and the normal production of white blood cells stops, leading to a loss of the body’s ability to fight disease [2]. Given that research has linked leukemia to genetics, one of the most important and accurate ways to diagnose and predict the disease is to use people’s DNA and genetic information [15].
 Following the argument that rs104893674 mononucleotide polymorphism is located within the exon and can directly increase or decrease gene function and consequently increase or decrease susceptibility to disease, the results of the present study are significantly associated with genotypic specificity and showed an allele for rs104893674 polymorphism with a risk of acute myeloid leukemia. It was found that people with AC genotype and “A” allele in this polymorphic position are more prone to acute myeloid leukemia. Therefore, considering the significant association of rs104893674 polymorphism with the risk of acute myeloid leukemia, it can be concluded that genetic markers are strong predictive strategies with high sensitivity for early detection of acute myeloid leukemia (Figure 1).


 

This study was ethically approved by the ethics committee of the Vice Chancellor for Research, Islamic Azad University, Kazerun Branch (Code: IR.IAU.KAU.REC.1398.051).

The present paper was extracted from the Msc. thesis of the first author, Department of Genetics, Faculty of Science, Kazerun Branch, Islamic Azad University.

All authors equally contributed to preparing this article.

The authors declared no conflict of interest.

The authors would like to thank the Vice Chancellor for Research of the Azad University, Kazerun Branch and all who helped with this research.