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Abstract Breast cancer is the most common female malignancy accounting for 22.9% and 37.7% of all female cancers worldwide and in Egypt, respectively. Breast cancer in Egypt carries an unfavorable prognosis with 29% mortality and 1: 3.7 incidences to mortality ratio.(1)) Breast cancer accounts for 16% of all cancer deaths among women globally, according to the report by the World Health Organization. It is the most common solid tumor diagnosed in women. Although the incidence of breast cancer increases with age, certain lifestyle and environmental factors play an important role on breast cancer risk. In Egypt, breast cancer is the most common cancer among women, representing 18.9% of total cancer cases (35.5% in women and 2.2% in men) among the Egypt National Cancer Institute (NCI) series of 10,556 patients during the year 2002-2003.(2) Breast cancer and cancer related diseases have been treated using surgery, chemotherapy, and radiation therapy, or a combination of these. But despite these therapeutic options, cancer remains associated with high mortality. This is basically due to difficulties in early diagnosis, exorbitant cost of treatment, with the often late presentation of breast cancer that generally characterizes cancer diagnosis among Egypt and other African women(4,5,10,28) Owing to these several shortcomings, there is a need for better therapeutic options which will increase the chances of survival of breast cancer patients with minimal or no side effects of treatment. Phytochemical prevention for severe health problems has recently gained scientific recognition worldwide. Studies on the pharmacological mechanisms and search for chemical structures of herbal extracts responsible for anticancer activity caught great interest(30-32) The aim of this work is to evaluate the caveolin 1 expression under the effect of Thuja occidentalis an ethanolic extract of Thuja occidentalis Linn. leaves and doxorubicin drugs in mice breast tissues induced by 7,12-dimethylbenzeneanthracene (DMBA) as well as biophysical and biochemical parameters..to fulfill this aim, 60 albino mice weighting 20-25 gm of 8-10 weeks of age were divided into two groups. The experimental groups received different concentrations of thuja with respect to the LD50 result as documented by Arthur et al.(31) Mice were treated with increasing doses of thuja. Uses of experimental animals in the study protocol were carried out in accordance with the ethical guidelines of the Medical Research Institute, Alexandria University (Appendix 2, Guiding Principles for Biomedical Research Involving Animals, 2011). group A: 10 Mice treated with distilled water only as a control group. group B: 50 Mice treated with 20 mg/ml/week of DMBA. This group was subdivided into five sub-groups; sub group B-1: 10 Mice treated with 20 mg/ml/week of DMBA only and were not receive any treatment, sub group B-2: 10 Mice treated with 10 mg/ml/day of thuja after DMBA administration, sub group B-3: 10 Mice treated with 7.5 mg/ml/day of thuja after DMBA administration, Sub group B-4: 10 Mice treated with 5 mg/ml/day of thuja after DMBA administration and sub group B- 5: 10 Mice treated with 5 mg/ml/day of doxorubicin after DMBA administration. For evaluation of the treatment effects to all studied groups the following investigations were done: 1- The length, the width and the height of the tumor were measured with a digital slide caliper every day before and after administration of different treatment modality and tumor volume was calculated. 2- Tumor mass inhibition ratio and tumor volume growth ratio were calculated. 3- Measurements of the dielectric properties of the tumor tissue and its surrounding at the end of treatment period to monitor the efficiency of the different treatment modality. 4- Analysis of toxic and trace metals: Analysis of the studied heavy metals and trace elements [lead (Pb), cadmium (Cd), chromium (Cr), nickel (Ni), iron (Fe), selenium (Se), copper (Cu) and zinc (Zn)] by Atomic Absorption. 5- Measurement of glutathione-S-transferase activity (GST), super oxide dismutase activity (SOD), glutathione reductase activity (GR), catalase activity (Cat), total antioxidant activity (TAC) and malondialdehyde (MDA). 6- Measurement the serum levels of ALT, AST, urea and creatinine were measured to detect the effect of thuja on liver and kidney functions. 7- Molecular detection of caveolin 1-mRNA gene expression by RT-PCR. 8- Evaluation of the structural changes induced by different treatment modality in tumor tissue using H&E stain by microscope. The experimental results showed that: 1) The antitumor effect of thuja increases with increasing of dose. Decreasing in the tumor volume, tumor growth rate and inhibition ratio were observed upon using increasing dose of thuja. However, the decline in the tumor volume has minimum value at 5 mg/kg/body weight thuja. 2) Relative permittivity of the tumor tissue decreased exponentially with increasing the administered dose of thuja. Relative permittivity of treated tumor tissue with different treatment modalities decreased than that of untreated tumor. The real conductivity increased exponentially with increasing applied dose of thuja for all the modes of treatments, while decreased in untreated tumor models. The variation of the imaginary permittivity with frequency showed sharp increase at low dose of thuja followed by exponential decreased with all methods of treatments used. 3) A significant decrease in both total body water and extracellular water percentage while a significant increase in intracellular water percentage of treated with 5mg thuja, 7.5mg thuja, doxorubicin and 10mg thuja respectively compared to untreated group. 4) A statistically significant elevation of heavy metals and trace elements (lead, cadmium, chromium, nickel and iron) concentrations was detected in breast tissue of mice having malignant breast tumors in comparison to control group. On the other hand, there is a significant decrease in the heavy metals and trace elements in the animals administered both thuja and carcinogen when compared with animals administered carcinogen alone. 5) All groups treated with DMBA, have a significant increase in the levels of MDA as compared with normal group animals. Animals in groups received thuja either simultaneously or post-treated exhibited significantly low levels of MDA, when compared with animals treated with DMBA only. 6) A decreased activities of enzymatic antioxidants (SOD, CAT, GR, GST and TAC) in comparison with normal animals. On the other hand, there is a significant increase in the enzymatic and non-enzymatic antioxidant guard in the animals administered both thuja and carcinogen when compared with animals administered carcinogen alone. 7) It was observed that treatment with thuja ameliorated the levels of serum creatinine and urea which is an indication of renal protection. This also confirms the protective role of thuja against DMBA-induced renal toxicity. Also treatment with thuja protected against increase in serum levels of ALT, AST, and GGT, which is an indication of hepatoprotection by thuja. This also confirms the protective role of thuja against DMBA-induced hepatotoxicity. 8) Caveolin 1 gene expression significantly higher in mice groups treated thuja extract (7.5 and 10 mg/kg body weight) than those treated with (5 mg/kg body weight) while the lowest expression was among untreated cancerous group. 9) The histological evaluation revealed that all tumors from the cancerous control group were highly malignant cells and none of the tumors showed necrosis. Tumors excised from animals receiving thuja extract (7.5 and 10 mg/kg body weight), significant areas of necrosis were present compared to (5 mg/kg body weight) treated group while in case of doxorubicin treated tumors, large foci of necrosis areas were present which were distinctly appeared. |