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Medical Journal of the Islamic Republic of Iran Volume 19 Number I Spring 1384 May 2005 ____ _ Ba _ s _ _ic S_ _ _i � ce i_ Ate_d_ __ __ __ � __ ____ __ __ __ __ ADENOSINE DEAMINASE ACTIVITY IN ESTROGEN RECEPTOR POSITIVE AND NEGATIVE HUMAN Downloaded from mjiri.iums.ac.ir at 20:38 IRDT on Friday May 5th 2017 BREAST CANCER CELL LINES MOHAMMAD HASHEMI,"1 FATEMEH KARAMI TEHRANI/ SAEID GHAVAMI, 1 AND MAJID SIRATI SABET.3 From the IDept. of Clinical Biochemistry, School of Medicine, Zahedan Medical University, Zahedan, iran, the 2Cancer Research Lab, Dept. of Clinical Biochemistry, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran, and the 3Dept. olClinical Biochemistry, School of Medicine, Qazvin Medical University, Qazvin, Iran. ABSTRACT Background: The aims of this study were to assay the activity of adenosine deami nase (ADA) in estrogen receptor positive (MCF-7) and negative (MDA-MB468) breast cancer cell lines. Methods: MDA-MB468 and MCF-7 breast cancer cell lines were cultured in complete medium, striped serum with and without 0.0 111M diethylstilbestrol (DES), complete medium in the presence and absence of 111M tamoxifen for 20 hr. Adenosine deaminase activity was determined using the colorimetric method described by Guisti and Galanti. Results: It was found that the activity of enzyme in estrogen receptor positive (ER+) cell line (MCF-7) was significantly higher than that of estrogen receptor negative breast cancer cell line (MDA-MB468). ADA activity in MCF-7 cells cultured in the presence of tamoxifen or charcoal-striped serum was significantly lower than that of control. Furthermore addition of diethylstilbestrol (DES) to the striped serum increased the value of ADA activity to that of control. Unlike MCF-7 cells, the activity of ADA in MDA-MB468 cells remained un changed upon treatment with tamoxifen or striped serum. Conclusion: These findings suggest estrogen responsiveness of ADA expression in MCF-7 cells. MJIRI, Vol. 19, No.1, 53-56, 2005. Keywords: Adenosine deaminase, breast cancer cell lines, estrogen receptor. INTRODUCTION loenzyme that catalyzes the deamination of adenosine and deoxyadenosine to inosine and deoxyinosine, re Adenosine deaminase (ADA, "Corresponding author: E.C.3.S.4.4) is a metal- Mohammad Hashemi, Ph.D., Dept. of Clinical Biochemistry, Zahedan Medical University, Zahedan, Iran. Tel.: 09173640366 spectively. I Due to the irreversibility of the reaction cata lyzed by ADA, this enzyme reaction seems to be one of the rate-limiting steps in adenosine degradation. ADA is present in virtually all human tissues, but the highest e-mail: mhdha [email protected] /53 Downloaded from mjiri.iums.ac.ir at 20:38 IRDT on Friday May 5th 2017 Downloaded from mjiri.iums.ac.ir at 20:38 IRDT on Friday May 5th 2017 ADAActivity in ER Positive and Negative Breast Cancer ity when compared to that of control, in contrast to the cer cells in tissue culture. Cancer Res 43: 4681-4690, 1983. reduce enzyme activity, although not at the same magni 11. Aitken SC, Lippman ME, Kasid A, Schoenberg DR: Rela tude as that observed for stripped serum. Furthermore, tionship between the expression of e strogen-regulated genes addition of DES, a potent synthetic estrogen, to the and estrogen-stimulated proliferation of MCF-7 m ammary stripped culture media stimulated ADA expression and tumor cells. Cancer Res 45: 2608-2615,1985 restored the activity. MDA-MB468 cells, however, ex hibited no changes in ADA activity upon treatment with Downloaded from mjiri.iums.ac.ir at 20:38 IRDT on Friday May 5th 2017 pyrimidine synthesis and utilization in human b r e ast can report of Xie et al,15 tamoxifen alone did significantly 12. Aitken SC, Lippman ME: Effect of est r o g e n and antiestrogens on growth-regulatory enzymes in h uma n tamoxifen of removing estrogen from the media. These breast cancer cells in tissue culture. Cancer Res 45: 1611- findings suggest E2 responsiveness of ADA expression 1620, 1985. in MCF-7 cells. In the MDA-MB468 cell line, the ADA 13. Cowan K, Levine R, Aitken S, G o l dsmith M, Douglass E, gene is not an E2 target gene and there must be other Clendeninn N, Nienhuis A, Lippman ME: Dihydr ofolate nuclear transcription factors to induce ADA expression. reductase gene amplification and possible rearrangement in It has been demonstrated that tamoxifen induces a estrogen-responsive methotrexate resistant human breast significant apoptosis in the MCF-7 cell line.23 One pos sible mechanism might be through the inhibition of ADA cancer cells. J BioI Chern 257: 15079-15086,1982. 14. Kasid A, Davidson NE, Gelmann EP, Lippman ME: Tran and subsequent accumulation of toxic adenosine and scriptional control of thymidine kinase gene expression by deoxyadenosine that causes inhibition of ribonucleotide estrogens and anti estrogens in MCF-7 human breast cancer reductase and also inactivation of s-adenosyl homocys teine hydrolase, which results in apoptosis. cells. J BioI Chern 261: 5562-5567,1986. 15. 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