|Year : 2015 | Volume
| Issue : 2 | Page : 341-344
Autoantibodies in the sera of breast cancer patients: Antinuclear and anti-double stranded DNA antibodies as example
Mohammed Elimam Ahamed Mohammed1, Khalid Abdelhafiz2
1 Scientific Services and Laboratories Unit, Nuclear Applications in Biological Sciences Institute (NABSI), Sudan Atomic Energy Commission, Khartoum, Sudan; Department of Chemistry, Faculty of Science, King Khalid University, Abha, Saudi Arabia
2 Scientific Services and Laboratories Unit, Nuclear Applications in Biological Sciences Institute (NABSI), Sudan Atomic Energy Commission, Khartoum, Sudan
|Date of Web Publication||7-Jul-2015|
Mohammed Elimam Ahamed Mohammed
Department of Chemistry, Faculty of Science, King Khalid University, Abha, Saudi Arabia
Source of Support: None, Conflict of Interest: None
Background: Inflammation and cell necrosis are one of the consequences that accompany breast cancer. However, inflammation and cell necrosis are well known to be involved in stimulation of cellular and humeral immunity.
Objectives: The aim of this study is to investigate the immune response to the inflammation that accompanies cancer through measuring plasma concentration of antinuclear antibodies (ANAs) and anti-double stranded deoxyribonucleic acid antibodies (ADSDAs).
Materials and Methods: Thirty-five newly diagnosed breast cancer patients were involved in this study from the Radiation Isotopes Center Khartoum (RICK) compared to 18 age- and sex-matched control subjects. Intravenous blood sample was obtained from each study subject and Enzyme Linked Immuno Sorbent Assay (ELISA) technique was used to determine the concentration of the two antibodies.
Results: Regarding the ANA concentration in the patients; the range was 0.7-1.8 IU/ml, mean was 0.96, and the standard deviation (SD) was 0.25; while the range of theconcentration in the control subjects was 0.3-0.6 IU/ml, mean was 0.47, and SD was 0.07. However, when the means of patients and controls were compared, the difference was significant (P < 0.000). Concerning the result anti-double stranded DNA (anti-dsDNA), its concentration range in the patients was 2.6-151.9 IU/ml, themean was 55.2, and SD was 25.6, while in healthy people concentration range was 26.1-97.3 IU/ml, the mean was 50.3, and SD was 16.9. There was no significant change between the patients and controls (P = 0.46).
Conclusion: The ANA concentration in the patients was significantly increased, while there was no significant difference between the results of ADSDAs in the patients and the control subjects.
Keywords: ADSDA, ANA, autoantibodies, breast cancer, cell necrosis
|How to cite this article:|
Mohammed ME, Abdelhafiz K. Autoantibodies in the sera of breast cancer patients: Antinuclear and anti-double stranded DNA antibodies as example. J Can Res Ther 2015;11:341-4
|How to cite this URL:|
Mohammed ME, Abdelhafiz K. Autoantibodies in the sera of breast cancer patients: Antinuclear and anti-double stranded DNA antibodies as example. J Can Res Ther [serial online] 2015 [cited 2020 Jul 10];11:341-4. Available from: http://www.cancerjournal.net/text.asp?2015/11/2/341/157314
| > Introduction|| |
An autoantibody is an antibody that is directed against one or more of the individual's own proteins. The causes of autoantibody production are varied and not well understood. However, excessive destruction of cells (necrosis and apoptosis) is major cause of autoantibody production. Autoantibodies are well known to be used for the classification of autoimmune diseases like the systemic lupus erythematosus (SLE),  also they are associated with infections and malignancies.  In recent years, a growing number of reports have showed that cancers may trigger production of autoantibodies, which may be used for early diagnosis. ,, Some studies showed that autoantibodies play a major role in angiogenesis and prognosis of cancer , and others stated that they can be used as targets for cancer drugs. 
Antinuclear antibodies (ANAs) are one of the autoantibodies which are synthesized in response to the contents of the cell nucleus. The contents of the cells and their nuclei appear in the blood because of apoptosis or cell necrosis. ANAs are used as a tool of diagnosis and classification of autoimmune diseases as SLE.  Recently, the ANAs are suggested to be used in the early diagnosis of breast cancer. 
Anti-double stranded deoxyribonucleic acidantibodies (ADSDA) are secreted in the blood in response to the presence of DNA molecules in the blood. As the ANA, the ADSDA is used in the diagnosis and classification of the SLE and other autoimmune diseases.  The ADSDA is a dual function protein since it hydrolyses extracellular DNA and it has a cytotoxic activity against tumor cells.  ADSDA was registered to be present in the sera of patients with SLE, rheumatological disorders, infections, and malignancy. 
This article hypothesized that ANA and ADSDA were significantly increased in the blood of newly diagnosed Sudanese breast cancer patients.
| > Materials and methods|| |
A total of 35 newly diagnosed breast cancer patients were involved in this study from the Radiation Isotopes Center Khartoum (RICK) in the period between December 2012 and February 2013. Eighteen healthy females were involved as control subjects and they were age- and sex-matched to the patients.
This study was implemented after academic approval from the institute of tropical diseases and after obtaining an ethnical license from the Sudan health authorities. The study subjects were involved after informed consent.
This study is descriptive, case-control, and hospital-based.
Three milliliter of intravenous blood sample was obtained from each study subject in plain tubes and the serum was allowed to separate, decanted in another tube, and kept at −20°C.
Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of ANA and ADSDA following the manufacturer (DRG) instructions.
However, the information of the ANA and ADSDA kits were DRG® ANA Profile-8 ELISA (EIA-4325) and DRG® Anti-dsDNA IgA (EIA-3565), respectively.
| > Results and discussion|| |
All the study subjects were females. The age range of the patients and controls was 30-65 years. Regarding the grade of the patients, there was no patient with grade I, 12 were with grade II, and 23 were classified as grade III. Concerning the stage of the patients, one patient was classified as stage I, nine as stage II, 20 as stage III, and five as stage IV.
The range of the ANA concentration in patients was 0.7-1.8 U/ml compared to 0.3-0.6 U/ml in control subjects, while the means were 0.96 and 0.47, respectively. Upon comparing the means of ANA in patients to control subjects the P - value was less than 0.000 [Figure 1]. All the patients were characterized by high ANA concentration compared to the control subjects (100%). The mean of ANA in the patients who are classified as grade II was 0.9 compared to the mean of the control group of 0.47; the difference was significant (P < 0.00) [Table 1]. Regarding patients with grade III, their mean ANA value was 1.0 compared to 0.47; which means that ANA is increased significantly in the patients with grade III (P < 0.00) [Table 1]. The ANA results of the patients with the different stages are presented in [Table 2]. However, the ANA value was significantly increased in patients with different stages compared to the control subject. P - values were less than 0.00.
|Figure 1: Mean values of ANA in patients and controls. The standard deviations of the control subjects and the patients were 0.07 and 0.25, respectively, and the difference between the two means was significant (P - value < 0.000). ANA = antinuclear antibody|
Click here to view
|Table 1: The mean values of ANA and ADSDA and the different grades of breast cancer|
Click here to view
|Table 2: Mean values of ANA and anti - ds DNA in the different stages of breast cancer|
Click here to view
Results of ADSDA
The range of ADSDA in the patients and controls were 2.6-151.9 and 26.1-97.3 U/ml, while the means were 55.2 and 50.3, respectively. By comparing the means, P - value was calculated as 0.46 [Figure 2]. Only one patient was with high anti-dsDNA concentration compared to the control subjects 2.9%. Concerning the ADSDA, its mean value in the patients with grade II was 68.4 compared to 50.2 of control subjects [Table 1]. From the above it was clear that ADSDA was not significantly increased in patients with grade II compared to the control subjects (P = 0.185) [Table 1]. The mean value of ADSDA in the patients with grade III was 51.2, while it was 50.2 in the normal subjects. However, it was clear that there was no significant difference between the patients and the controls [Table 1]. Regarding the results of ADSDA in the different stages, it was clear that there was no significant difference in all the stages compared to the control [Table 2].
|Figure 2: Mean values of ADSDA in patients and controls. The standard deviation of the control subjects was 16.9 and of the patients was 25.6. There was no significant difference between the two study groups (P -value = 0.46). ADSDA = Anti-double stranded deoxyribonucleic acid antibody|
Click here to view
| > Discussion|| |
The results of this study showed that the ANA concentration was significantly increased in the patients compared to the control subjects. All the patients were with high ANA concentration compared to the control subjects. These results may be due to increased cancerous cell death, which leads to exposure of nuclear material to the immune system.
Dlair and colleagues studied different types of cancers including breast (20 female cases), prostate (12 male cases), gall bladder (nine females), urinary bladder (four males), and other cancers (four males and one female). From the 20 breast cancer patients, he found that ANA was positive in nine patients (45%).  Another study showed that 20 out of 50 cancer patients were characterized by high titer of ANA.  In a mini review article, Wasserman et al., (1976)  stated that, the previous studies showed contradicting results ranging from high frequency of ANA expression to null expression in breast cancer patients. However, Wasserman et al., expressed that the ANA has prognostic significance. In a previous study associated with effect of tamoxifen on the blood level of ANA, Mohammed Mohammed and his colleagues showed that newly diagnosed breast cancer patients are associated with negative ANA values and that the tamoxifen treatment significantly increased the blood ANA concentration.  Shukaili and his research group found that ANA was significantly increased in the blood of Omani gastric cancer patients. 
Regarding the ADSDA results, we have shown that there was no significant variation between the breast cancer patients and the control subjects and that only one patient has high ADSDA concentration (2.9%). In a case study report and unlike our finding, Turgutalp et al., found that prostate cancer and acute kidney injury induced high expression of ADSDA.  It has been known that all cancers exert positive ADSDA titer. The patients with high expression of ADSDA has better prognosis due to the anti-tumor activity of the ADSDA autoantibodies.  Similar to our findings, but in gastric cancer, Shukaili et al., found that the ds DNA autoantibodies were negative in the patients and the controls. 
| > Conclusions|| |
This study concluded that:
- The ANA blood concentration was significantly increased in all breast cancer patients irrespective of the grade or stage, that is, it was increased in all the stages and grades. The ANA test may be used in the early diagnosis of breast cancer
- There was no significant difference between the concentration of ADSDA in patients and controls. The ADSDA test is not useful in the early diagnosis of breast cancer.
In order to generalize the findings of this study, it is better to be repeated with large sample size.
| > References|| |
Haugbro K, Nossent JC, Winkler T, Figenschau Y, Rekvig OP. Anti-dsDNA antibodies and disease classification in antinuclear antibody positive patients: The role of analytical diversity. Ann Rheum Dis 2004;63:386-94.
Attar SM, Koshak EA. Medical conditions associated with a positive anti-double-stranded deoxyribonucleic acid. Saudi Med J 2010;31:781-7.
Turgutalp K, OÐuz EG, Akca M, Kiykim A. Anti-dsDNA positivity in a patient with prostate cancer and acute kidney injury: A case report. Turk Neph Dial Transpl 2013;22:221-3.
Al-Shukaili A, Al-Jabri AA, Moundhri MS. Prognostic value of auto antibodies in the serum of Omani patients with gastric cancer. Saudi Med J 2006;27:1873-7.
Lacombe J, Mangé A, Solassol J. Use of autoantibodies to detect the onset of breast cancer. J Immunol Res 2014;2014:574981. Available from: http://dx.doi.org/10.1155/2014/574981
[Last accessed on 2015 Mar 30].
Liu BC, Dijohnson DA, O′Rourke DJ. Antibody profiling with protein antigen microarray in early stage cancer. Expert Opin Med Diagn 2012;6:187-96.
Ferna´ndez Madrid F. Autoantibodies in breast cancer sera: Candidate biomarkers and reporters of tumorigenesis. Cancer Lett 2005;230:187-98.
Kelly-Worden M, Hammer L, Gebhard R, Schrader L, Griffin M, Cooper D, et al
. Anti-nuclear antibodies positive serum from systemic lupus erythematosus patients promotes cardiovascular manifestations and the presence of human antibody in the brain. J Pharm Bioallied Sci 2014;6:198-204.
Imran A, Neelam F, Tariq M. Incidence of circulating antinuclear antibodies in cancer patients. Indian J Med Sci 2003;57:113-6.
Pavlovic M, Kats A, Cavallo M, Chen R, Hartmann JX, Shoenfeld Y. Pathogenic and Epiphenomenal Anti-DNA Antibodies in SLE. Autoimmune Dis 2010;2011:462841.
Mohammad DO, Najim RH. Relation of anti-nuclear antibodies with different types of cancer. Tikrit J Pharma Sci 2011;7:51-4.
Editorial: Autoantibodies in breast cancer. Br Med J 1976;1:244.
Mohammed Mohammed EA, Sharief Nawahil A, Abukashawa Sumaia MA. Tamoxifen- induced lupus Erythematosus. J Drug Metab Toxical 2013;4:1000138. Available from: http://dx.doi.org/10.4172/2157-7609.1000138
[Last accessed on 2015 Mar 30].
Qinghua C, Wei X, Zhenke W, Lin X, Kang L, Yiwei C, et al
. An anti-double-stranded DNA monoclonal antibody induced by tumor cell-derived DNA inhibits the growth of tumor in vitro
and in vivo
via triggering apoptosis. DNA Cell Biol 2008;27:91-100.
[Figure 1], [Figure 2]
[Table 1], [Table 2]