Serum ceruloplasmin, copper, zinc levels and Cu/Zn ratio in malignancy Dr. Lamia M. Al-Namaa Dr. Jamal A. Abdul Barry Dr. Zinab A. Al-Manaseer Dr. Hashim S. Al-Khayat ALTOMA

Introduction Introduction Malignant cells characterizes by hyper metabolic activities and high proliferation rates leading to increased production of harmful waste materials like free radicals. A balance between oxidant carcinogens and endogenous antioxidant defense is of particular relevance to the carcinogenesis

Introduction, cont. Introduction Neoplastic diseases activate antioxidant defense systems. As a result, concentrations of antioxidant enzymes and their co-factor elements appear to change.

Introduction, cont. Introduction The levels of essential trace metals zinc (Zn), copper (Cu) and ceruloplasmin (copper transport protein) have been found to be critical parameters.

Introduction, cont. Introduction Our interest in these two trace metals because it was establish that they are involved in immunological and inflammatory reactions. In addition they have been recognized to play an important role as cofactors of superoxide dismutase (SOD) an important antioxidant enzymes which are used to dismutate the H2O2.

Introduction, cont. Introduction Ceruloplasmin (Cp) is a copper-carrying glycoprotein (carries up to 90% of Cu in plasma). It uses ferric oxidase activity to prevent the occurrence of toxic Fe products. In addition, it controls membrane lipid oxidation. As an acute phase reactant, ceruloplasmin was found to be increased in gastrointestinal (GIT) cancers . In fact, it is claimed to be a prognostic and diagnostic factor in various malignancies.

Introduction, cont. Introduction Gastrointestinal and breast cancers are frequent diseases in our localities. With respect to patient’s outcome or care; surgeons always look forwards for better patients management and follow-up.

Aims The present study was undertaken to: Estimate the serum level of trace elements Zinc, copper and its related protein ceruloplasmin in patients with those malignant disease. And calculate their Cu/Zn ratio. Compared the results obtained with that from non-malignant subjects. Find which of the above parameters is a better indicator of the antioxidative status in malignant patients

Patients and Methods A prospective study, conducted through a period of 8 months, from October 2000 to May 2001, during which 116 patients whom were admitted to the Surgical ward of Basrah General Hospital were studied.

Patients and Methods Individuals investigated were: 33 patients with cancer of gastrointestinal tract (GIT), aged 25 - 65 years 16 patients with breast cancer, aged 32 –70 years 67 age and sex- matched non malignant person ( suffering from surgical conditions other than malignancy) served as controls. From each person the following information were recorded : age, sex, family history, past history of other diseases, smoking, medication, and dietary habits. Person with DM, HT, IHD were exculded

Methods Venous fasting blood sampled was collected from each subject before operation. Serum levels of copper and zinc were measured by atomic absorption spectrophotometry. Ceruloplasim activity was estimated enzymatically according to the procedure of Houchin’ method.

RESULTS and DISCUSSION

Table 1: Characteristic of malignant and non-malignant subjects Malignant Cases Variables Control (67) Breast (16) G I T (33) 37 (55%) (45%) 9 (56.3%) 7 (43.7%) 10 (30 %) 23 (70 %) < 45 > 45 Age 33 (49%) (41%) 16 (100%) 16 (48%) 17 (42%) Female Male Sex 12 (17.9%) 55 (82.1%) 1 (6.25%) 15 (93.75%) 10 (30%) 23 (70%) +ve -ve Smoking habit 4 (6%) 63 (94%) 7 (43.75%) 9 (56.25%) 3 (10%) 30 (90%) Family history

Table-2: Serum level of Cp, Cu, Zn and Cu/Zn ratio in malignant and non-malignant subjects Type of malignancy Breast (16) G I T (33) Non-Malignant (67) Parameters 69.3 + 18.4 ** 65 + 19.2 ** 52 + 17 Ceruloplasmin 98.2 + 24.6 * 90.5 + 20.7 84.9 + 23.3 Copper 87 + 23.9 82.9 + 27 ** 97.9 + 21 Zinc 1.18 + 0.49 *** 1.25 + 0.49 *** 0.9 + 0.28 Cu/Zn ratio Values were expressed as mean ± SD, (n) * P= 0.05, **P<0.05, ***P <0.01 All results were expressed as mg/dl.

The possible cause for ceruloplasmin elevation was related to incraese hepatic synthesis.

Zinc deficiency often results in elevated blood levels of copper, due to the dynamic competition of these metals in the body. Clinical observations and research indicate the copper/zinc ratio appears to be more important than either copper or zinc levels alone. The ratio represents the distance between benign and malignant tumors.

Table-3: Serum level of Cp, Cu, Zn and Cu/Zn ratio in GIT and Breast malignant cases and non-malignant controls in relation to age   Age (yr) Parametermg/dl Non- malignant Mean + SD (n) G I T cancer Breast Cancer Cp 51.7 + 3 (37) 70.6 + 17.3 **(10) 70.6 + 26.9 **(9) < 45 Cu 81.6 + 4(37) 92.3 + 17.8 *(10) 89 + 18.4 *(9) Zn 95.8 + 3.7 (37) 98.6 + 32.5 (10) 90.5 + 19.3(9) Cu/Zn 0.91 + 0.07(37) 1.23 + 0.5* (10) 1.24 + 0.2 * (9) 50.9 + 3 (30) 64.15 + 8.5 **(23) 63.6 + 10.3 **(7) > 45 92.37 + 4.6 (30) 92.6 + 24.5(23) 102.5 + 24.5 (7) 98 + 4.39 (30) 78.6 + 24.7 *(23) 86.5 + 30.7 (7) 0.96 + 0.8(30) 1.3 + 0.39 *(23) 1.3 + 0.6 *(7) *p <0.05, **p<0.01

Table-4: Serum level of Cp, Cu, Zn and Cu/Zn ratio in GIT and Breast malignant cases and non-malignant controls in relation to gender   Sex Parametermg/dl Non- malignant Mean + SD (n) G I T cancer Breast Cancer Cp 54.7 ± 16.2 (33) 61.2 ± 17 ** (16) 69.3 ± 18.4 *** (16) Female Cu 88.14 ± 21.7 (33) 87.47 ± 17.3 (16) 98.4 ± 25.16 * (16) Zn 100.7 ± 18.4 (33) 73.8 ± 24.3** (16) 87 ± 23.9 (16) Cu/Zn 0.92 ± 0.3 (33) 1.27 ± 0.6** (16) 1.18 ± 0.4 *** (16) 46.4 ± 15.9 (34) 69.3 ± 20.7*** (17) - Male 82.4 ± 21.4 (34) 94 ± 26 (17) 90.27 ± 20.7 (34) 88 ± 27 (17) 0.97 ± 0.41 (34) 1.27 ± 0.56* (17) *p = 0.05 **p <0.05, ***p<0.01

Table-5: Serum level of Cp, Cu, Zn and Cu/Zn ratio in GIT and Breast malignant cases and non-malignant controls in relation to smoking habits   Smoking Habit Parameter mg/dl Non- malignant Mean + SD (n) G I T cancer Breast Cancsr Cp 52.25 ± 15.2 (12) 76 ± 15.47 * (10) 63 (1) + ve Cu 82 ± 29.4 (12) 97.5 ± 29.4* (10) 46 (1) Zn 103 ± 27 (12) 92.5 ± 27.7 (10) 85 (1) Cu/Zn 0.84 ± 0.4 (12) 1.2 ± 0.58* (10) 0.54 (1) 52.1 ± 17.6 (55) 79.6 ± 19 *(23) 67.5 ± 20* (15) - ve 85.6 ± 22 (55) 87.3 ± 17 (23) 99.8 ± 22* (15) 96.8 ± 19.6 (55) 77.6 ± 27** (23) 87 ± 25 (15) 0.92 ± 0.33 (55) 1.28 ± 0.6* (23) 1.23±0.46 * (15) *p <0.05, **p<0.01

Table-3: Serum level of Cp, Cu, Zn and Cu/Zn ratio in GIT and Breast malignant cases and non-malignant controls in relation to family history   Family History Parametermg/dl Non- malignant Mean + SD (n) G I T cancer Breast Cancsr Cp 47.8 ± 10.6 (4) 54 ± 13.07 (3) 62.3 ± 10.9 (7) + ve Cu 76.6 ± 28.5 (4) 89 ± 25 (3) 98 ± 26 (7) Zn 105 ± 9.8 (4) 84.5 ± 32 (3) 91.7 ± 29 (7) Cu/Zn 0.732 ± 0 .2 (4) 1.2 ± 0.73 (3) 1.12 ± 0.4 (7) 52.5 ± 17.5 (63) 66 ± 18.9* (30) 71.6 ± 4.6*(9) - ve 85.5 ± 23 (63) 90.6 ± 22 (30) 95 ± 26 (9) 97.5 ± 21.7 (63) 82.2 ± 27.8* (30) 83.4 ± 20 (9) 0.90 ± 0.35 (63) 1.25 ± 0.59* (30) 1.23± 0.56 (9) *p <0.05, **p<0.01

Conclusions 1. In this preliminary study, the level of antioxidant (ceruloplasmin) and enzymes cofactors ( Cu and Zn) were altered in patients with G I T and breast malignancy as compared with those non-malignant cases. These changes were with high significant increase in the level Cp and Cu/Zn ratio.

Conclusions 2. Measurement of Cp and estimation of Cu / Zn ratio is a better indicator of antioxidant status in malignant patients

Recomandation: Suggested future work to: Support a role for Cp and Cu/Zn ratio as a diagnostic tool in different malignancy Estimate the level of Cp, Cu, Zn and Cu/Zn ratio in malignant patients post operatively which may help in prognosis of disease. Establish a correlation between these parameters and the severity of the disease ALTOMA

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