مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Estimation of ALP, GPT and GOT Activities in Iraqi Patients Female With B reast Cance r Z. I. AL-Mashhadani , A. J. A. Muk hlis, * A. S. A-Razaq AL-Faraji Departme nt of Chemistry, College of Education I bn- ALhaitham, Unive rsity of Baghdad. *Center for Market Rese arch & Consume r Protection, Unive rsity of Baghdad Received in : 24 July 2011 Accepte d in : 18 October 2011 Abstract To invest igate the activity and role of certain enzy me markers in 30 p atients female with breast cancer (non-treated, treated, and treatment with recovered).The serum activity of enzy me tumor markers (ALP, GPT and GOT) of (30) p atients with breast cancer, and (7) healthy control subjects by using st atist ical analysis: There is significant difference higher in activity of serum enzy me tumor markers (ALP, GPT, and GOT) in all patients as comp ared with healthy control. Key word: Breast Cancer, Enzy me tumor markers. Introduction Breast Cancer: Cancer is a d isease in whi ch cells beco me abnormal and form more cells in an uncontrolled way [1]. With breast can cer, the cancer begins in cells that make up the breast (usually in the tubes that carry milk to nipple or the glands that make milk). The can cerous cells for m a mass of tissue called a malignant tumor that st arts from cells in the breast . Sometimes, the cancer sp reads to other p arts of the body [1,2]. The disease occurs mostly in women, but men can get breast cancer as well. Breast cancer is the most common cancer amon g women, other than skin cancer. It is the se cond leading cause of cancer death in women, after lung can cer[1-4]. About one in eight women will be d iagnosed with breast cancer during their lif etime. Breast cancer also st rikes men but in much lower numbers (1 in 100)[4]. There are several types of breast cancer, although some of them are quite rare. In some cases a single breast tumor can have a combination of these typ es or have a mixture of invasive and in situ cancer which are[5]: 1- Ductal carcino ma in situ. (DCIS). 2- Lobular carcino ma in situ. (LCI S). 3- Invasive (or infiltrating) ductal carcinoma. (IDC). 4- Invasive (or infiltrating) lobular carcinoma. (ILC). Alkal ine phosphatase (ALP) (Ec: 3.1.3.1): Alkaline p hosp hatase (ALP) is ahydrolase enzy me resp onsible for removing p hosp hate group from many ty p es of molecules, including nucleotides, p roteins, and alkaloids. The process of removing the phosp hate group is called dep hosp hory lation[6]. As the name suggests, alkaline p hosp hatases are most effective in an alkaline environment. It is sometimes used sy nony mously as basic p hosp hatase[7]. مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 In humans, alkaline p hosp hatase (ALP) is found in many tissues, including bone, liv er, intest ine, kidney , and p lacenta [8]. High (ALP) usually means that the bone or liver been damaged[9]. The normal range is 20 t o 140 IU/L[10]. Alani ne Transami nase (ALT) or (GPT) (Ec: 2.6.1.2): Alanine transaminase or ALT is a transaminase enzy me. It is also called seru m glutamic p y ruvic transaminse (SGPT) or alanine aminotransferase (ALAT). ALT is found in serum and in var ious bodily tissues, but is most commonly associated with the liver. It catalyzes the two p arts of t he alanine cycle[11]. Significantly elevated levels of ALT often suggest the existence of other medical p roblems such as viral h epatitis, congestive heart failure, liver dama ge, b ile duct p roblems, infectious mononucleosis, or my op athy [11]. Reference ran ge of ALT : 6-37 U/L[10]. Aspartate Transami nse (AS T) or (GOT) (Ec: 2.6.1.1): Asp artate transaminase (AST) also called serum glutamic oxaloacetic transaminase (SGOT) or asp artate aminotransferase (ASAT/AAT /Asp AT ) is similar to alanine transaminase (ALT) in that it is another enzy me associated with liver p arenchymal cells[12]. ALT is found p redominately in the liver, with lesser quantities found in the kidneys, heart, and skeletal muscle. As a result the ALT is a more sp ecific indicator of liver inflammation than the AST, as the AST may also be elevated in diseases affecting other organs, such as the heart or muscles in myocardial infarction, also in acute p ancreatitis, acute hemolytic anemia, severe burns, acute renal disease, musculoskeletal diseases, and trauma[13]. AST (SGOT) is commonly measured clinically as a p art of diagnost ic liver function tests, t o determine liver health. Reference ranges of AST:6-34 IU/L[14]. The aim of our st udy is to evaluate some biochemical p arameters like enzy mes as tumor biomarker (i.e. ALP, GPT, GOT). In Iraqi p atients suffering from breast cancer and comp ared that with t he same parameters in normal healthy Iraqi control. Material and Methods Patie nts: Blood samples were collected from a thirty breast cancer (females) (Non- treated (8) female, treated (12) female, treated and r ecovered (10) female), their age was range (19-60) y ears at the M edical C ity Hospital in Baghd ad. Seven app arently healthy individuals (females) were selected with age range (20-43) years. Ten milliliters (ml) of venous blood were collected into p lain tubes from each p atient and healthy individuals after 12 hours fast. The blood samples were allowed to st and for 15 minutes (min) then centrifuged at 3500 rp m for 10 min. Serum was frozen at -20 º C till used for the est imation of ALP, GPT, GOT . De termination of Some Enz yme Marke rs: De termination of Alkaline Phosphatase (ALP Activity): Colorimetric determination[15,16] of the ALP activity which reaction scheme is as follows: Phe nyl phospha te A lka line Phosphata se Phenol + Phosphate Free phenol librated by hy drolysis of t he subst rate reacts then with 4-amino-antip y rine in the p resence of alkaline p otassium ferricyanide to form a red-coloured comp lex which absorbance measured at 510nm is directly p rop ortional to t he ALP activity in the sp ecimen. Sodium arsenate in corporated in the reagent abolishes further enzy me activity and p revents t he dilution of the colour inherent in earlier methods. مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Calculation ( kind and k ing u nits/1 00ml) . - . = 20 . standard A bs Assay A bs specime n blank ALP activity A bs  De termination of Aminotransfe rases (ALT) or (GPT) Activity: Colorimetric determination[17-19] for GPT activity according to the following reactions: GPT Alanine ketoglutarate Pyruvate glutamate    The p y ruvate formed is measured in its derivated form, 2,4-dinitrop henylhydrazone at 505nm. Calculation number of GPT units/ml in serum were calculated using the standard curve. De termination of Aminotransfe rases (AS T) or (GOT) Activity: Colorimetric d etermination[17-19] of GOT activity according to the followin g reactions: GOT L A spartate ketoglutarate Ox aloacetic L glutamate      The oxaloacetate formed is measured in its derivated form, 2,4- dinitrop heny lhydrazone at 505nm. Calculation The number for GOT units/ml in serum were calculated using the standard curve. Results and Discussion Results Se rum Enz yme Marke rs Levels: Se rum Alkaline Phosphatase (ALP) Acti vity: Data in Table (1) and Fig. (1) show that, t he mean ± SD of ALP activity in serum of control was (16.6565 ± 5.7695 IU/L). While the mean ± SD of ALP activity in serum of p atient with breast cancer ( No treatment) was (38.4583 ± 27.4304IU/L) and for (treatment and treatment & recancer) were (56.8010 ± 23.6085IU/L) and (47.6151 ± 29.0696IU/L) resp ectively. The results showed no significant differ ence (P  0.05) in serum of ALP activity of Breast Cancer p atient's no treatment, treatment and treatment & recancer as comp ared with healthy control. مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Fig.(1) of our results shows that no treatment breast cancer p atients female was two times high in ALP activity , while treatment p atients high four times and treatment with recancer was t hree times high activity than healthy control. S erum Alani ne Transami nase (ALT) or (GPT) Activity: Table (2) and Fi g. (2) shows t hat, the Alanine transaminase (ALT) or (GPT) activity in serum of control is (19.2500 ± 2.4749 units/ml). While the (ALT) activity in serum of breast cancer p atient no treatment is (24.7000 ± 13.9803 un its/ml) and treatment and treatment & recancer p atient's breast cancer wer e (28.9143 ± 22.0185 units/ml) and (21.8200 ± 12.4333 units/ml) resp ectively. The results showed that there is no si gnificant difference (P  0.05) in serum ALT activity of Breast Cancer patient comp ared with that of healthy control. Our results from the Fig.(2) show that no treatment was high one time in ALT activity , while treatment female Breast cancer patients was t wo t imes high er in activity , and treatment with recancer half time high er than healthy control. S erum Aspartate Transami nase (AS T) or (GOT) Activity Data in Table (3) and Fig. (3) show that the mean ± SD of asp artate transaminase (AST) or (GOT) activity in serum of control was (28.75 ± 3.8891units/ml). While the mean ± SD of AST activity in serum of p atient with breast cancer (No treatment, treatment, and treatment & recancer) were (38.3667±17.5342units/ml), (30.0750 ± 5.6076 units/ml), and (50.6167 ± 25.6691units/ml) resp ectively. The results showed no significant differ ence (P  0.05) in serum of AST activity of all breast cancer p atients as comp ared with healthy control. Our st udy from the Fig.(3) shows that t he no treatment p atients of breast cancer was high er AST activity one time than the control and high in treatment, while in treatment with recancer is higher two times comp ared with healthy control. Discussions Alkaline phosphatase (ALP) Activity: Our results demonst rate that none of our p atients with total-ALP activities lower than the control p resented metastases. On the contrary , p atients with bone metastases show a significantly increased total-ALP activity . This finding is in agreement with data from other authors[20]. It has been demonst rated that total-ALP is highly sensitive to detect hepatic metastases[20]. Our p atients with hepatic or bone metastases show the highest total-ALP activity and there are no significant differences of total-ALP activity between them. According to our results, total-ALP could not differentiate between hepatic and bone metastases and it is necessary to take into account bone-ALP values that, in bone metastases[21]. Alani ne transami nase (ALT) or (GPT) Activity: Alanine transaminase was sign ificantly high in patients with lymp h node metast asis at d is eas e p r es en t at io n as co mp ar ed t o co nt ro l. T h is s u gges t s t h e r o le o f alan in e aminotransferas e in relation to axillary lymp h node metastasis and disease p rognosis. The p athogenesis of drug induced liver disease usually involve the p articip ation of the parent drug or its metabolites t hat either directly effect t he cell biochemistry or elicit an immune resp onse. Susceptibility to drug induced h epatot oxicity is also influenced by genetic and environmental ris k f act ors. Unp red ict able, low fr equency, iod iosy ncr ati c r eact ions o ften o ccur on a back ground of a h igher rate of mild asy mptomatic liver injury , it is very difficult to detect t hem b ut t h ey may b e d et ect ed by mon it or in g s eru m amin otr ans fer as e lev els [2 2 ] . مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Some trials suggest biochemical evaluation as the bett er tool for the screening of metastatic disease at the time of diagnosis of breast cancer[23,24]. In these trials ALT is suggested as first -line examination to detect liver or bone metastases. Aspartate Transami nase (AS T) or (GOT) Activity: Asp artate transaminase (AST) is familiar markers of liver function and to ascertain the non-involvement of sy st emic toxicity , the activities of marker enzy me like Asp artate transaminase (AST) was assay ed. In our st udy , the AST levels is higher in cancer when comp ared to normal control[24]. As a marker for liver metastases in breast cancer p atients and also as a marker for hepatot oxicity , asp artate transaminase was found to be increased. It was concluded that the elevations in the activities could be due to the decreased sy nthesis of degradative p roducts which could have resulted in the elevation in the circulation. Elevations in liver transaminases following tamoxifen administ ration have been observed. Tissue damage is the sensitive feature in the cancerous conditions so any deterioration or destruction of the membrane can lead to the leakage of these enzy mes from the tissues. Hence elevation of these liver sp ecific enzy mes observed in breast cancer condition may be due to the progression of tumor growt h[25]. Conclusion Our results suggest that ALP, GPT, and GOT may p lay an imp ortant role in breast cancer. A significant elevation in the activity of ALP, GPT, and GOT in female breast cancer. Recommendation Although, some of our data were statistically no significant, our interp retation from thes results indicate that the chemotherapy are unusefull in treatment of breast cancer. Re ferences 1. Singh, S.P., (2008). Text book of Biochemistry , 4 th Ed., CBS p ublishers & Dist ributors, New Delhi, Bangalore. 2. Harris, L.; Fritsche, H.; M ennel, R.; (2007). American society of clinical Oncology 2007 up date of recommendations for the use of tumor markers in breast cancer. J. clin. Oncol., 25:5287-5312. 3. Foubert, E.; Craene, B. and Berx, G.(2010) The Snail 1- Twist 1 Consp iracy in malignant breast Cancer progression. Breast Cancer Research, 12:206. 4. 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Philadelp hia, W.B., Saunders, P: 1395-1457. مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 9. Posen, S. and Doherty , E. (2010). The measurement of serum alkaline p hosp hates in clinical medicine. Adv. Clin. Chem., 22:165. 10. Bishop , M .L.; Fody , E.P.and Schoeff, L.E. (2010). Clinical chemist ry , techniques, p rinciples, correlations by lip p incott Williams & Wilkins. 11. Paul, T.and Giboney M .D. (2010). M ildly Elevated liver Transaminase levels in the Asy mptomatic Patient. American Family p hy sician. 12. Almo, S.C.; Smith, D.L.; Danishefsky , A.T .and Ringe, D. (M arch 1994). “ The structural basis for the altered substrate sp ecificity of the R292D actine site mutant of asp artate aminotransferase from E.Coli ”. Prot ein Eng. 7(3):405-12. 13. Lott , J.A. and Stang, J.M . (2010). 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Table(1): Alkaline phosphatase (ALP) Activity (IU/L) in serum of Patients with Breast Cancer (no treatment, treatment, and treatment & recancer) and healthy control. Groups N Mean SD t-test No treatment 8 38.4583 27.4304 P  0.05 Treatment 12 56.8010 23.6085 P 0.05 Treatment & recancer 10 47.6151 29.0696 P  0.05 Control 7 16.6565 5.7695 P< 0.05 مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Table(2): Alanine transaminase (ALT) or (GPT) Activity (units/ml) in serum of Patie nts with Breast Cancer (no treatment, treatment, and treatme nt & recancer) and healthy control. Table(3): Aspartate transami nase (AS T) or (GOT) Activity (units/ml) in se rum of Patients with Breast Cancer (no treatment, treatment, and treatme nt & recancer) and healthy control. Groups N Mean SD t-test No treatment 8 38.3667 17.5342 P  0.05 Treatme nt 12 30.0750 5.6076 P 0.05 Treatme nt & reccancer 10 50.6167 25.6691 P  0.05 Control 7 28.7500 3.8891 P< 0.05 Groups N Mean SD t-test No treatment 8 24.7000 13.9803 P  0. 05 Treatme nt 12 28.9143 22.0185 P 0.05 Treatme nt & recancer 10 21.8200 12.4333 P  0. 05 Control 7 19.2500 2.4749 P< 0.05 Fig. (1): Alkaline phosphatase (ALP) Activity (IU/L) in se rum of Patients with Breast Cancer (no treatment, treatme nt, and treatment & recancer) and healthy control. مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Fig.(2):Alanine transaminase (ALT) or (GPT) Acti vity (units/ml) in serum of Patie nts with Breast Cancer (no treatment, treatment, and treatme nt & recancer) and he althy control. Fig.(3):Aspartate transami nase (AS T) or (GOT) Activity (units/ml) in serum of Patients with Breast Cancer (no treatme nt, treatment, and treatme nt & recancer) and he althy control. مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 في GOT و GPT واالمینوترانسفیرز ALPتقییم الفوسفات القاعدي ت المصابات بمرض سرطان الثديالمریضات العراقیا ،عبد الجبار عبد القادر مخلص ،زهیر ابراهیم المشهداني * علیاء سعدون عبد الرزاق الفراجي جامعة بغداد، ابن الهیثم –كلیة التربیة ، قسم الكیمیاء جامعة بغداد، مركز بحوث السوق وحمایة المستهلك * 2011 تشرین األول 18:حث في قبل الب2011 تموز 24:استلم البحث في الخالصة .لغرض بیان دور الفعالیة وبعض األنزیمات الواسمة في المرضى المصابین بسرطان الثدي فعالیــة تقیـسو). بعـالج مـع رجـوع المـرض، بعـالج ، بـدون عـالج (مـصابة بـالمرض ) 30( عینـات تتـألف مـن ت اخـذ نـساء ) 7(مریـضة و ) 30(فـي مـصل ) GOT و GPT واالمینـو ترانـسفیرز ALPعدي الفوسفات القا(األنزیمات الواسمة كانت نتـائج البحـث للمجـامیع المـذكورة باسـتخدام التحلیـل اإلحـصائي . مجموعة سیطرة خالیة من أي مرضبوصفهن تطوعن فـي جمیــع المرضــى مقارنــة (GOT, GPT, ALP)حــدوث ارتفـاع معنــوي فــي فعالیــة األنزیمـات الواســمة : يأتكمـا یــ . باألصحاء .أنزیمات واسمة ورمیة ، سرطان الثدي : الكلمات المفتاحیة مجلة إبن الھیثم للعلوم الصرفة و التطبیقیة 2012 السنة 25 المجلد 1 العدد Ibn A l-Haitham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012