Full Text:   <3048>

CLC number: R73-3

On-line Access: 2013-01-08

Received: 2012-01-11

Revision Accepted: 2012-06-29

Crosschecked: 2012-11-29

Cited: 1

Clicked: 5018

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2013 Vol.14 No.1 P.40-46


Binding of circulating autoantibodies in breast cancer to native and peroxynitrite-modified RNA

Author(s):  Sheereen Tarannum, Zarina Arif, Khursheed Alam

Affiliation(s):  Department of Biochemistry, Faculty of Medicine, Aligarh Muslim University, Aligarh 202002, India; more

Corresponding email(s):   kalam786@rediffmail.com

Key Words:  RNA, Peroxynitrite, ELISA, Breast cancer, Band shift assay

Sheereen Tarannum, Zarina Arif, Khursheed Alam. Binding of circulating autoantibodies in breast cancer to native and peroxynitrite-modified RNA[J]. Journal of Zhejiang University Science B, 2013, 14(1): 40-46.

@article{title="Binding of circulating autoantibodies in breast cancer to native and peroxynitrite-modified RNA",
author="Sheereen Tarannum, Zarina Arif, Khursheed Alam",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Binding of circulating autoantibodies in breast cancer to native and peroxynitrite-modified RNA
%A Sheereen Tarannum
%A Zarina Arif
%A Khursheed Alam
%J Journal of Zhejiang University SCIENCE B
%V 14
%N 1
%P 40-46
%@ 1673-1581
%D 2013
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1200015

T1 - Binding of circulating autoantibodies in breast cancer to native and peroxynitrite-modified RNA
A1 - Sheereen Tarannum
A1 - Zarina Arif
A1 - Khursheed Alam
J0 - Journal of Zhejiang University Science B
VL - 14
IS - 1
SP - 40
EP - 46
%@ 1673-1581
Y1 - 2013
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1200015

peroxynitrite (ONOO) is a powerful oxidant and nitrosative agent and has in vivo existence. The half life of ONOO at physiological pH is less than 1 s. It can react with nucleic acids, proteins, lipoproteins, saccharides, cardiolipin, etc., and can modify their native structures. Action of ONOO, synthesized in the authors’ laboratory by a rapid quenched flow process, on structural changes of commercially available RNA was studied by ultraviolet (UV), fluorescence, and agarose gel electrophoresis. Compared to native RNA, the ONOO-modified RNA showed hyperchromicity at 260 nm. Furthermore, the ethidium bromide (EtBr) assisted emission intensities of ONOO-modified RNA samples were found to be lower than the emission intensity of native RNA-EtBr complex. Agarose gel electrophoresis of ONOO-modified RNA showed a gradual decrease in band intensities compared to native RNA, an observation clearly due to the poor intercalation of EtBr with ONOO-modified RNA. Native and ONOO-modified RNA samples were used as an antigen to detect autoantibodies in sera of patients with clinically defined breast cancer. Both direct binding and inhibition enzyme-linked immunosorbent assay (ELISA) confirmed the prevalence of native and 0.8 mmol/L ONOO-modified RNA specific autoantibodies in breast cancer patients. Moreover, the progressive retardation in the mobility of immune complexes formed with native or 0.8 mmol/L ONOO-modified RNA and affinity purified immunoglobulin G (IgG) from sera of breast cancer patients supports the findings of the direct binding and inhibition ELISAs. The peroxynitrite treatment to RNA at a higher concentration appears to have damaged or destroyed the typical epitopes on RNA and thus there was a sharp decrease in autoantibodies binding to 1.4 mmol/L ONOO-modified RNA. It may be interpreted that cellular nitrosative stress can modify and confer immunogenicity on RNA molecules. Higher concentrations of nitrogen reactive species can be detrimental to RNA. However, the emergence of native as well as 0.8 mmol/L ONOO-modified RNA as a novel antigen/substrate for autoantibodies in breast cancer patients indicates that, in future, these molecules might find a place on the panel of antigens for early diagnosis of breast cancer.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Ali, R., Alam, K., 2002. Evaluation of Antibodies against Oxygen Free Radical-Modified DNA by ELISA. In: Armstrong, D. (Ed.), Methods in Molecular Biology: Oxidative Stress Biomarkers and Antioxidant Protocols. Humana Press, New Jersey, p.171-181.

[2]Burney, S., Caulfield, J.L., Niles, J.S., Tannenbaum, S.R., 1999. The chemistry of DNA damage from nitric oxide and peroxynitrite. Mutat. Res., 424(1-2):37-49.

[3]Chapman, C., Murray, A., Chakrabarti, A., Thorpe, C., Woolston, U., Sahins, A., Robertson, J., 2007. Autoantibodies in breast cancer: their use as an aid to early diagnosis. Ann. Oncol., 18(5):868-873.

[4]Denicola, A., Souza, J.M., Radi, R., 1998. Diffusion of peroxynitrite across erythrocyte membrane. PNAS, 95(7):3566-3571.

[5]Dixit, K., Khan, M.A., Sharma, Y.D., Moinuddin, Alam, K., 2011. Peroxynitrite-induced modification of H2A histone presents epitopes which are strongly bound by human ant-DNA autoantibodies: role of peroxynitrite-modified-H2A in SLE induction and progression. Hum. Immunol., 72(3):219-225.

[6]Goding, J.W., 1978. Use of staphylococcal protein A as an immunological reagent. J. Immunol. Methods, 20:241-253.

[7]Goldstein, S., Lind, J., Merenyi, G., 2005. Chemistry of peroxynitrites as compared to peroxynitrates. Chem. Rev., 105(6):2457-2470.

[8]Hughes, M.N., Nicklin, H., 1968. The chemistry of pernitrites. Part I. Kinetics of decomposition of pernitrous acid. J. Chem. Soc. A, 15:450-452.

[9]Ischiropoulos, H., Zhu, L., Beckman, J.S., 1992. Peroxynitrite formation from macrophage derived nitric oxide. Arch. Biochem. Biophys., 298(2):446-451.

[10]Jin, H., Zangar, R.C., 2009. Protein modifications as biomarkers in breast cancer. Biomark Insights, 4:191-200.

[11]Khan, M.A., Dixit, K., Jabeen, S., Moinuddin, Alam, K., 2009. Impact of peroxynirite modification on structure and immunogenicity of H2A histone. Scand. J. Immunol., 69(2):99-109.

[12]Koppenol, W.H., Kissner, R., Beckman, J.S., 1996. Syntheses of peroxynitrite: to go with the flow or on solid grounds. Methods Enzymol., 269:296-302.

[13]Mantovani, A., Allvena, P., Sica, A., Balkwill, F., 2008. Cancer-related inflammation. Nature, 454:436-444.

[14]Masuda, M., Nishono, H., Ohshima, H., 2002. Formation of 8-nitroguanosine in cellular RNA as a biomarker of exposure to reactive nitrogen species. Chem. Biol. Int., 139(2):187-197.

[15]Niles, J.C., Wishnok, J.S., Tannenbaum, S.R., 2006. Peroxynitrite-induced oxidation and nitration products of guanine and 8-oxoguanine: structures and mechanisms of product formation. Nitric. Oxide, 14(2):109-121.

[16]Pacher, P., Beckman, J.S., Liudet, L., 2007. Nitric oxide and peroxynitrite in health and disease. Physiol. Rev., 87(1):315-424.

[17]Pope, S., Land, J.M., Heales, S.J.R., 2008. Oxidative stress and mitochondrial dysfunction in neurodegeneration: cardiolipin a critical target. Biochim. Biophys. Acta, 1777(7-8):794-799.

[18]Sing, R.K., Sudhakar, A., Lokeshwar, B., 2011. From normal cell to malignancy: distinct role of proinflammatory factors and cellular redox mechanisms. J. Cancer Sci. Ther., 3(4):70-75.

[19]Tan, E.M., Zang, J., 2008. Autoantibodies in tumor-associated antigens: reporters from the immune system. Immunol. Rev., 222(1):328-340.

[20]Wasserman, J., Glas, U., Blomgren, H., 1975. Autoantibodies in patients with carcinoma of the breast. Correlation with prognosis. Clin. Exp. Immunol., 19(3):417-422.

[21]Yu, H., Venkatarangan, L., Wishnok, J.S., Tannenbaaum, S., 2005. Quantitation of four guanine oxidation products from reaction of DNA with varying doses of peroxynitrite. Chem. Res. Toxicol., 18(12):1849-1857.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE