Synthesis and Characterization of Complex for Anti-Breast Cancer, activity evaluation on Cell Line MCF-7 and Study of some Biochemical Parameters in Women with the disease

Abstract

Cancer is one of the most common cause of death throughout the world. Breast cancer, is the most common cancer among the females and the major reason of mortality in western countries. In Iraq it is reported as the second reason of death after cardiovascular diseases. This study included 140 samples (50 samples from patients newly diagnosed with breast cancer, 50 samples others have taken chemotherapy and 40 samples from healthy women ) . The age of the selected cases including the control group was rang from (30-70) years In this work derivative of L-ascorbic acid (Vitamin C ) had been synthesized. and was reacted with cis-[PtCl2(NH3)2] to give complex with molecular formula cis-[Pt(Asc)(NH3)2]. The complex was characterized by FT-IR and 1HNMR spectrum . The High Content Screening (HCS) assay of 400μg/ml, 200μg/ml, 100μg/ml, 50μg/ml and 25μg/ml from complex cis-[Pt(Asc)(NH3)2] showed toxic effects toward Michigan Cancer Foundation-7(MCF-7)cell line after 24 hours of treatment in dose dependent manner with a reduction in the number of total cell count, reduction in the mitochondrial membrane potential (MMP), increased in the nuclear intensity, increase in the membrane potential and increase in cytochromeC. Furthermore, the study covered the effect of age and BMI on several parameters including hormones such as estradiol , progesterone and prolactin, aromatase, alkaline phosphatase and reduced glutathione , lipid profile , vitamin C, and some elements such as Selenium (Se) , Zinc (Zn+2) , Magnesium (Mg+2 ), and Calcium (Ca+2) . The main results of the previous section showed a significant increase in the level of estradiol, progesterone, prolactin, aromatase, alkaline phosphatase, cholesterol, triglyceride, low density lipoprotein , very low density lipoprotein and calcium in serum of breast cancer in comparasion with healthy control . While the II following parameters showed a significant decreased in the level of such high density lipoproteins, reduced glutathione(GSH), vit C, Total Antioxidant Capacity, as well Selenium, Zinc, Magnesium in serum of breast cancer compared with the case control study. In addition, this study includes determination of the activity of alkaline phosphatase followed by isolation and purification and an estimation of molecular weight, effecting of some factors on enzyme activity and studying the chemical kinetic study of alkaline phosphatase. Optimization of the extraction and purifying the alkaline phosphatase (ALP) from serum was achieved by the following steps of purification: Firstly, the precipitation; Using ammonium sulphate in saturated degree (60 %) showed increase in specific activity of partial purification ALP enzyme from (0.0021U/mg ) in crude sample to (0.0031U/mg) after first step of purification . Second, using ion-exchange chromatography; with DEAE-cellulose to isolate three types of isoenzymes. The partial purification of the tissue – nonspecific (TNSALP) isoenzyme(І) was carried out using Sephadex G100 as gel - filtration technique , and sodium dodecyl sulfate-polyacryl amide gel electrophoresis ( SDS-PAGE ) which gave only one single band . On the other hand, the molecular weight of pure isoenzyme(І) using PAGE revealed single band with molecular weight of 97 KD. The kinetic study of the purified isoenzyme (І) was carried out, which was found that the optimum concentration for the substrate disodium phenyl phosphate (DPP) was (5mM) and the Km was (1.4)mM , while its optimum pH was 10 , and optimum temperature was 37o C . After studying the effect of the reaction time on the enzymatic activity , it was found that the maximum activity is after 15 minutes of incubation . The study also included the effect of some metal ions ,amino acids and complex cis-[Pt(Asc)(NH3)2] on the purified isoenzyme . The results revealed activation effect of Co+2 , Mg+2 , Ni+2 , while Zn+2 , Cu+2 , EDTA and the amino acids such as Cysteine , Leucine , Phenylalanine , Tryptophan , as well the III complex cis-[Pt(Asc)(NH3)2] showed an inhibition effects on activation of ALP. The complex of cis-[Pt(Asc)(NH3)2] acted as non-competitive inhibitor . Finally, the correlation relationship between alkaline phosphatase activity and the levels of some parameters has been represented by the following result: The positive correlation between the enzyme activity and T.C, T.G, LDL-C, Ca levels. While there was a negative correlation with of HDL-C, GSH, Vit C, Se, Zn, and Mg