In acute lymphoblastic leukemia (ALL), the bone marrow loses its ability in differentiation and aamaturation of blood cells at different stages. ALL is a malignancy, which is characterized by clonal proliferation and accumulation of neoplastic cells (
Faderl et al., 1998). The disease is divided into two categories of B cell and T cell. B cell is the most common subtype in children and adults (
Hoffman et al., 1991).
Cancer and its treatments can affect the biological functions and change the nutritional status of patients (
de Carvalho et al., 2011). Zinc and copper are important cofactors for several enzymes that can play an important role in maintaining the integrity of DNA. Zinc and copper also act as antioxidants (
Demir et al., 2011) and zinc may reduce the duration and intensity of fibril neutropenia and its associated complications (
Radhakrishnan et al., 2013). Changes in serum levels of zinc and copper have been found in lymphoproliferative disorders in addition to the breast cancer, lung, and gastrointestinal tumors (
Avisar et al., 2012,
Shils and Shike, 2006). Experimental data support the presence of slight zinc deficiency in malignancies (
Sgarbieri et al., 2006). Zinc deficiency may cause disturbances in oxidation, mitochondrial function, DNA repair, and cancer induction and progression. Impaired zinc metabolism in pathogenesis of leukemia was identified in 1949. Zinc seems to improve the overall ability of patients in resisting the toxic side effects of chemotherapy (
Zuo et al., 2006).
Copper is a necessary element for various metalloenzymes including ceruloplasmin, cytochrome oxidase, and dopamine hydroxylase. In normal conditions, 96% of serum copper is in the ceruloplasmin. Ceruloplasmin is a serum glycoprotein with 8 copper atoms and 12 sialic acid chains per molecule, required for oxidation of Fe
+2 to Fe
+3 for storage in the form of ferritin or being transmitted via transferrin. If catabolism of ceruloplasmin depends on cutting two sialic acids by neuraminidase, any phenomenon that reacylates the cut points, inhibits the catabolism of ceruloplasmin. Thus, the increase of serum copper observed in malignancies is due to the reduction of ceruloplasmin’s catabolism. Another speculation is that the balance between serum sialic acid and ceruloplasmin is due to reduced activity of neuraminidase or increased concentration of sialic acid. This increase of acylation or decrease of deacylation occurs in the liver (
Fisher and Shifrine, 1978). In cancer, we have oxidative conditions in body that can cause oxidative damage to lipids and production of malondialdehyde (MDA). MDA is a mutagenic compound in mammalian cells and can have reaction with DNA bases such as guanine (G), adenine (A), cytosine (C), and resulted in the production of M1G (Malodialdehyde deoxy Guanosin) M1A (Malodialdehyde deoxy Adenine), and M1C (Malodialdehyde deoxyCitosine), which can cause damage to DNA (
Valko et al., 2006). Studies showed that hs-CRP level is positively associated with cancer and in the recent studies there was a positive relation between elevated hs-CRP and risk of incident of any type of cancer. There are two hypotheses about increasing hs-CRP in cancer; the first one is that, increasing in hs-CRP level is a result of underlying cancer and the second believes that because of oxidative stress in cancer, inflammation can initiate carcinogenesis by inactivating mutations in tumor-suppressor genes or post translational modifications in proteins involved in DNA repair or apoptosis control (
Lee et al., 2011). To our knowledge, there are not any data about evaluating serum levels of vitamin D in these patients before and after chemotherapy. In the present study, the aim was to compare serum levels of zinc, copper, vitamin D, and inflammatory status after eight courses of chemotherapy.
Materials and Methods
Study design and participants: In this before and after study, After confirmation of the Ethics Committee of Tehran University of Medical Sciences, during two years (2014-2016), 30 patients (17 men and 13 women) with ALL, who referred to Taleghani hospital participated in this study (leukemia was diagnosed through bone marrow aspiration and the cells were classified according to the French-American-British). Chemotherapy was done based on HYPERCVAD protocol for these patients. In the first visit of patient for chemotherapy, 10 mL of venous blood was taken prior to treatment. Blood samples were then centrifuged for 20 minutes at 400 g. At the end, the plasma was transferred to another tube. All samples were stored at -80°C. Taking blood samples were repeated at the end of the eighth chemotherapy courses.
Measurements: Serum concentration of zinc was measured by ZellBio GmBh (V4126) kit that is made by Germany and (nm 546 calorimeter) based on the directions of kit producer corporation. The human prospective value is usually 72.6-127 µg/dL (11.1-19.5 µmol⁄L) for men and 70-114 µg/dL (10.7-17.5 µmol/L) for women, with the sensitivity of 10 µg⁄dL.
Serum copper was measured by ZellBio GmBh (V4126) kit that is made by Germany and (nm 5u0-590 calorimeter) based on the directions of kit producer corporation. This kit could measure copper in serum in the range of 70-140 µg/dL (11-22 µmol⁄L) for men and 80-155 µg/dL (12.6-24.4 µmol⁄L) for women, with the sensitivity of 1 µg⁄dL.
Vitamin D was measured by Calbiotech (cat. No: VD220B) kit that is made by Canda, based on ELISA method, according to the directions of kit producer corporation. According to this kit, values of vitamin D were defined as bellow: <10 ng/mL deficient, 10-30 ng/mL inadequate, 30-100 ng/mL adequate, and >100 ng/mL toxic. The sensitivity of this kit was 67.0 ng⁄mL.
hs-CRP was measured by Canadian DBC ( diagnostics Biochem Canada) kit (cat. No: CAN-CRP-4360) based on ELISA method and following the directions of kit producer corporation. This kit could measure hs-CRP 132-9710 ng/mL for men and 139-6578 ng/mL for women with a sensitivity of 10 ng/mL.
ZellBio GmB, (CAT No.ZB-MDA96A), performed measurement of serum MDA (CAT No.ZB-MDA 48A) that are made by Germany via calorimeter and according to the manufacture's user guide. This kit was created by using a combination of MDA-TBA formed by the reaction of MDA and thiobarbituric acid (TBA) under a high temperature operation. This kit was capable of measuring MDA in the range of 0.78-50 μM and its sensitivity was 0.1 μM.
Data analysis: In this study, data were analyzed using SPSS version 12. At first, data distribution was checked by Kolmogorov-Smirnov test; in the case of normal distribution, paired samples t-test was used to compare the means before and after chemotherapy, and if the distribution of the data was not normal, Wilcoxon test was applied to compare before and after means. In addition, p-value < 0.05 was considered statistically significant.
Ethical considerations: Tehran University of Medical Sciences ethic committee approved this study.
Results
The mean of hematological variable before and after of chemotherapy are summarized in Table 1. Results showed that the serum concentration of zinc at the end of 8th chemotherapy course raised significantly and serum copper decreased significantly in these patients. In addition, our result showed significant decrease in serum concentration of MDA and vitamin D at the end of eight course of chemotherapy, whereas serum concentration of hs-CRP showed no significant difference. Our result showed that before chemotherapy 63.3% of ALL patients had insufficient serum vitamin D whereas all the patients had insufficient serum vitamin D after 8th course of chemotherapy.