Obese mammary tumour-bearing mice are highly sensitive to doxorubicin-induced hepatotoxicity

Abstract

Background: Breast cancer is a major health burden for women, worldwide. Lifestyle-related risk factors, such as obesity and being overweight, have reached epidemic proportions and contributes to the development of breast cancer. Doxorubicin (DXR) is a chemotherapeutic drug commonly used to treat breast cancer, and although effective, may cause toxicity to other organs. The mechanisms and effects of DXR on hepatic tissue, and the contributing role of obesity, in breast cancer patients are poorly understood. The aim of this study was therefore to investigate the effects of doxorubicin on hepatic tissue in an obese tumour-bearing mouse model. Methods: A diet-induced obesity (DIO) mouse model was established, where seventy-four three-week-old female C57BL/6 mice were divided into two main groups, namely the high fat diet (containing 60% kcal fat) and standard diet (containing 10% kcal fat) groups. After eight weeks on their respective diets, the DIO phenotype was established, and the mice were further divided into tumour and non-tumour groups. Mice were subcutaneously inoculated with E0771 triple negative breast cancer cells in the fourth mammary gland and received three doses of 4 mg/kg DXR (cumulative dosage of 12 mg/kg) or vehicle treatments via intraperitoneal injection. The expression levels of markers involved in apoptosis and alanine aminotransferase (ALT) were compared by means of western blotting. To assess the pathology and morphology of hepatic tissue, haematoxylin and eosin staining was performed. The presence of fibrosis and lipid accumulation in hepatic tissues were assessed with Masson’s trichrome and Oil Red O staining, respectively. Results: Our western blot results indicated that a significant increase in the ratio of cleaved caspase-8 and caspase-8 protein expression was observed in the standard diet tumour-bearing mice treated with DXR compared to the high fat diet tumour-bearing mice treated with DXR. Microscopic examination of liver tissue showed significant changes in the high fat diet tumour-bearing mice treated with DXR, consisting of macrovesicular steatosis, hepatocyte ballooning and lobular inflammation, compared to the standard diet tumour-bearing mice treated with DXR and the control group (standard diet mice). These changes are the hallmarks of non-alcoholic fatty liver disease, associated with obesity.Conclusion: Our results suggest that DXR activated the extrinsic apoptotic pathway in tumour-bearing mice on the standard diet. The histopathological findings indicated that DXR caused significant hepatic parenchymal injury in the obese tumour-bearing mouse model. Hepatotoxicity is aggravated in obesity as an underlying co-morbidity. It has been shown that obesity is associated with poor clinical outcomes in patients receiving neo-adjuvant chemotherapy treatment regimens.