الفهرس 
INTRODUCTION AND AIM OF STUDYOnly 14 pages are availabe for public view
 |  | from | 107 |  |  |
| | | from | 107 | | |
Abstract
BC is the most common cancer in women, where accounts for more than one out of every ten new cancer diagnosed cases each year. The most conventional therapeutic strategies for malignant tumors involve surgery, chemotherapy and radiotherapy. Radiotherapy is toxic and damage adjacent normal cells in addition to malignant cells. Hence, seeking for new radiosensetizing agents to prevent or decrease the side effects and complications of radiotherapy is of a central importance.
The aim of the work was to evaluate the radiosensitization of artesunate by targeting stemness and hypoxia markers on solid ehrlich tumor implanted in mice. This study included 64 Ehrlich solid carcinoma (ESC) bearing mice divided into these groups: group I (Control): Include 8 untreated ESC bearing mice. group II (IR): Include 8 ESC bearing mice were treated with ionizing radiation alone (6Gy). group III (ART 50 mg/kg/day): Include 8 ESC bearing mice were treated with artesunate
alone (50mg/kg/day). group IV (IR& ART 50 mg/kg/day): Include 8 ESC bearing mice were treated with the combination of ionizing radiation (6Gy) and artesunate (50mg/kg/day). group V (ART100 mg/kg/day): Include 8 ESC bearing mice were treated with artesunate alone (100 mg/kg/day). group VI (IR& ART 100 mg/kg/day): Include 8 ESC bearing mice were treated with the combination of ionizing radiation (6Gy) and artesunate (100 mg/kg/day). group VII (ART 200 mg/kg/day): Include 8 ESC bearing mice were treated with artesunate alone (200mg/kg/day). group VIII (IR & ART 200 mg/kg/day): Include 8 ESC bearing mice were treated with the combination of ionizing radiation (6Gy) and artesunate (200mg/kg/day).
At the end of the experiment, the body weight, tumor volume, and tumor inhibition rate were calculated, and this was followed by :
Stemness markers CD44 and ALDH1A1 concentrations in tumor tissue homogenate were assayed by ELISA technique.
Hypoxia markers HIF-1α and VEGF were assayed by ELISA technique.
P53 was be stained by immunohistochemistry technique
Histopathological study: tumor tissue was excised, fixed in 10 % neutral formalin, embedded in paraffin blocks and sectioned. Tissue sections were stained with H&E stain and examined using light microscope.
Chapter (6): Summary, Conclusions & Recommendations
59
Our results showed that:
Artesunate and ionizing radiation alone or in combination with ionizing radiation has a potential synergistic cytotoxic and suppressive effect against ESC bearing mice. These results are confirmed by significant increase in tumor inhibition rate in ESC bearing mice treated with artesunate, ionizing radiation alone or in combination of both.
Different doses of artesunate in combination with ionizing radiation significantly increase tumor damage reflected by increase tumor inhibition rate in comparison to radiation only treated ESC-bearing mice. This result demonstrate that artesunate enhances the radiosensitization in the animal model.
The maximum tumor inhibition rate was produced at dose 200 mg/Kg of artesunate in combination with ionizing radiation. These findings were Furthermore confirmed by the histopathological examination of solid tumor with H&E staining where the more pronounced effect in tumor regression was showed with Artesunate 200 mg/kg in combination with ionizing radiation.
Artesunate and ionizing radiation alone or in combination has apoptotic effect against tumors in ESC bearing mice in a dose dependent manner, where a gradual significant increase was found in the apoptotic cells number following treatment of ESC bearing mice with artesunate and radiation alone or in combination when comparing with that that of untreated ESC-bearing mice. on comparing with ESC bearing mice treated with ionizing radiation, the number of apoptotic cells in ESC bearing treated with 50 mg/kg of artesunate was insignificant difference, whereas the apoptotic cells in those treated with artesunate at doses 100 mg/kg and 200 mg/kg was significantly higher. Moreover, our finding showed that different doses of artesunate in combination with ionizing radiation significantly increase apoptotic cells number in comparison to radiation only treated mice. These results indicating that, artesunate can induce apoptosis in the tumor and enhance apoptotic effect of ionizing radiation in a dose dependent manner.
Our data confirmed apoptotic effect against tumors where a gradual significant increase was demonstrated in the expression of p53 following treatment of ESC bearing mice with artesunate and radiation alone or in combination when comparing with that of untreated ESC-bearing mice. significant elevation was found in the expression of p53 in the ESC bearing mice treated with 200 mg/kg of artesunate in combination with ionizing radiation when comparing to those treated with 200 mg/kg of artesunate. These results indicate the maximum expression of p53 was produced in ESC bearing mice treated dose of 200 mg/Kg of artesunate in combination with ionizing radiation. Suggesting the radiosensitivity effect of artesunate is P53 dependent.
The statistical analyses of our data revealed that insignificant difference in all comparisons with respect to the tissue homogenate HIF-1α, but the statistical analyses of VEGF revealed that significant elevation in the levels of VEGF in ESC bearing mice treated with artesunate alone or in combination with ionizing radiation in comparison to ESC bearing mice treated with ionizing radiation alone. These results give evidence that, artesunate induces the hypoxia and this effect is often temporary.
The statistical analyses of our results revealed that insignificant difference in the concentration of CD44 and ALDH1A1 between the ESC bearing mice treated with ionizing radiation and control ESC bearing mice. ESC bearing mice treated with artesunate alone or in combination with ionizing radiation showed insignificant difference when compared to control ESC bearing mice, but Administration of artesunate at high concentration 200mg/kg alone or in combination with ionizing radiation was significantly higher than in control ESC-bearing mice or ESC bearing mice treated with ionizing radiation only. These results indicated that, antitumor and radiosensitivity effect of artesunate are stemness independent.