Effect of recombinant human erythropoietin and doxorubicin in combination on the proliferation of MCF-7 and MDA-mb231 breast cancer cells

Patients with cancer often exhibit signs of anemia as the result of the disease. Thus, cancer chemotherapies often include erythropoietin (EPO) in the regime to improve the survival rate of these patients. The aim of the present study was to determine the effect of EPO on doxorubicin-treated breast cancer cells. The cytotoxicity of doxorubicin alone or in combination with EPO against the MCF-7 and MDA-MB 231 human breast cancer cells were determined using an MTT cell viability assay, neutral red (NR) uptake assay and lactate dehydrogenase (LDH) assay. The estimated half maximal inhibitory concentration values for doxorubicin and the combination of doxorubicin with EPO were between 0.140 and 0.260 µg/ml for all cells treated for 72 h. Treatment with doxorubicin in combination with EPO led to no notable difference in cytotoxicity, compared with treatment with doxorubicin alone. The antiproliferative effect of doxorubicin at a concentration of 1 µg/ml on the MDA MB 231 cells was demonstrated by the decrease in viable cells from 3.6x10(5) at 24 h to 2.1x10(5) at 72 h of treatment. In order to confirm apoptosis in the doxorubicin-treated cells, the activities of caspases-3/7 and 9 were determined using a TBE assay. The results indicated that the activities of caspases-3/7 and 9 were significantly elevated in the doxorubicin-treated MDA-MB-231 cells by 571 and 645%, respectively, and in the MCF 7 cells by 471 and 345%, respectively, compared with the control cells. EPO did not modify the effect of doxorubicin on these cell lines. The results of the present study suggested that EPO was safe for use in combination with doxorubicin in the treatment of patients with breast cancer and concurrent anemia

Treatment of MCF-7 and MDA-MB-231 human breast cancer cell lines with erythropoietin, doxorubicin and their combination

The cancer chemotherapies are formulated to target the rapidly proliferating cancer cells more efficiently than those normal cells of low proliferating rate. In cancer therapies erythropoietin (EPO) is often used in combination with chemotherapeutic drugs to treat the cancer-associated anemia. In this study, it is hypothesized that EPO does not modify the cytotoxic effect of doxorubicin (DOX). Thus the objective of the study was to determine the effect of DOX-EPO combination treatment on human breast cancer cell lines. The cytotoxicity of DOX (1 μg/mL) alone or in combination with EPO (1 μg/mL DOX – 1 IU/mL EPO) against two human breast cancer cell lines, the MCF-7 and MDA-MB-231 cell, was determined by 3-4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide cell viability assay, neutral red uptake and lactate dehydrogenase assays. The activity caspases and morphology of the treated cells were also determined. Doxorubicin inhibited MCF-7 cells with IC50 of 0.217±0.05 and 0.127±0.01 μg/mL when determined by the MTT and NR assay respectively, at 72 hours.On the MDA-MB-231 cells the IC50 of DOX was 0.12±0.09 μg/mL and 0.118±0.04 μg/mL by the MTT and NR assay, respectively, at 72 h of treatment. Doxorubicin in combination with EPO did not exhibit any notable difference in cytotoxicity to the cell lines when a compared to DOX alone. The anti-proliferative effect of DOX alone was obvious on the MCF-7 and MDA-MB-231 cell, showing decline in cell counts from 27.4 and 27.3 × 105 to 2.1 and 7.0 × 105 cells respectively, after 72 h of treatment. Light microscopic examination of DOX-treated MCF-7 and MDA-MB-231 cells at 72 h demonstrated apoptotic changes in cellular morphology characterized by cell rounding followed by a loss of adherence with subsequent cell shrinkage and blebbing. The mechanism of cell death was determined through the caspases-3 and -9 activities of the treated cells. The results demonstrated that caspases-3 and-9 activities were significantly (p<0.05) elevated early in DOX-treated MCF-7 MDA-MB-231 cells after 24 h, suggesting the apoptotic effect of DOX is via the mitochondrial pathway. Erythropoietin did not cause any change in caspase level in the treated cells. This study shows that EPO did not modify the cytotoxic effect of DOX on MCF-7 and MDA-MB-231 cells and thus is safe to be used with DOX in the treatment of breast cancers in patients with concurrent anemia

001 Egyptian consensus-based recommendations for the diagnosis and targeted management of Kawasaki disease. An initiative by the Egyptian College of Pediatric Rheumatology

Abstract Background Kawasaki disease (kDa) is a self-limiting acute vasculitis that affects small and medium-sized vessels, and is the most common cause of acquired heart disease in children. It is also an important reason for long-term cardiac disease into adulthood. Rapid diagnosis and management of kDa is challenging due to the heterogeneity of the disease, yet is vital for improving outcome. To date, there are no Egyptian nationally agreed, evidence-based guidelines concerning the diagnosis and management of kDa in children. Consequently, treatment regimens vary broadly. Objectives To develop a consensus, evidence-based recommendations for the diagnosis, evaluation and management of children living with kDa. Methods This study was carried out to achieve an Egyptian expert consensus on a management strategy for kDa using Delphi technique. A multistep process strategy was adopted, which started by developing 16 key clinical questions by scientific committee according to the Patient/Population, Intervention, Comparison, Outcomes and Time (PICOT) approach. The core leadership team identified clinicians and researchers with expertise in pediatric rheumatology all over Egypt. An evidence-based, systematic, literature review was conducted to compile evidence for the kDa management. Delphi process was implemented (3-rounds) to reach a consensus. Results Twenty-five expert panel participated in the 3 rounds with response rate 100%. A total of 21 recommendations, categorized into 9 domains (Definition, disease activity, predicting the development of coronary disease, assessment and monitoring (lab, imaging), treatment (acute and after acute attack), management of resistant cases, management of complications (cardiac complications, MAS and arthritis), vaccination and long term follow up. The Agreement with the recommendations (rank 7–9) ranged from 83.6–95.7%. The Consensus was reached (i.e. ≥75% of respondents strongly agreed or agreed) on all the clinical standards. Algorithm for management has also been developed. Conclusion This was an expert, consensus recommendations for the diagnosis and treatment of kDa, based on best available evidence and expert opinion. The recommendations provided a management approach based on easy-to-use algorithm and with the support of complementary tests. The implication to policy, practice, research and advocacy to provide updated recommendations for better management of kDa

43 Egyptian evidence-based consensus recommendations for diagnosis and targeted management of childhood-onset systemic lupus erythematosus. An initiative by the Egyptian College of Pediatric Rheumatology

Abstract Background Childhood-onset systemic lupus erythematosus (cSLE) is a rare prototype of a multi-systemic, inflammatory, heterogeneous and potentially life-threatening autoimmune condition with quite significant linked morbidity. To date, there are no Egyptian nationally agreed, evidence-based guidelines concerning the diagnosis and management of kDa in children and treatment is often based on clinical expertise. Consequently, treatment regimens vary broadly. This work was an initiative by the Egyptian College of Paediatric Rheumatology aiming at optimising the management approaches for children and young adults with cSLE. Objective To provide evidence-based consensus recommendations for diagnosis and management of cSLE Methods This study was carried out to achieve an Egyptian expert consensus on a management strategy for cSLE using Delphi technique. A multistep process strategy was adopted, which started by developing 18 key clinical questions by scientific committee according to the Patient/Population, Intervention, Comparison, Outcomes and Time (PICOT) approach. The core leadership team identified clinicians and researchers with expertise in pediatric rheumatology all over Egypt. An evidence-based, systematic, literature review was conducted to compile evidence for the cSLE management. Delphi process was implemented (3-rounds) to reach a consensus. Results Twenty-five expert panel participated in the 3 rounds with response rate 100%. A total of 24 recommendations, categorized into 14 domains (Targeted population, criteria for classification, definition of disease activity status, definition of flare up, lab tests, treatment targets, monitoring, treatment, recommendations for specific organ system, prognostic markers, refractory condition, vaccination and transition program. The Agreement with the recommendations (rank 7–9) ranged from 84.8–94.8%. The Consensus was reached (i.e. ≥75% of respondents strongly agreed or agreed) on all the clinical standards. Algorithm for management has also been developed. Conclusion This work provided an updated management approach for cSLE patients. This evidence-based informed consensus process is expected to support uniform, high quality standards of care for children with cSLE in Egypt The implication to policy, practice, research and advocacy: to provide updated recommendations for better management of cSLE