Anti-Angiogenic Treatments Interact with Steroid Secretion in Inflammatory Breast Cancer Triple Negative Cell Lines

Abstract

Human inflammatory breast cancer (IBC) is a highly angiogenic disease for which antiangiogenic therapy has demonstrated only a modest response, and the reason for this remains unknown. Thus, the purpose of this study was to determine the influence of different antiangiogenic therapies on in vitro and in vivo steroid hormone and angiogenic growth factor production using canine and human inflammatory breast carcinoma cell lines as well as the possible involvement of sex steroid hormones in angiogenesis. IPC-366 and SUM149 cell lines and xenotransplanted mice were treated with different concentrations of VEGF, SU5416, bevacizumab and celecoxib. Steroid hormone (progesterone, dehydroepiandrostenedione, androstenedione, testosterone, dihydrotestosterone, estrone sulphate and 17β-oestradiol), angiogenic growth factors (VEGF-A, VEGF-C and VEGF-D) and IL-8 determinations in culture media, tumour homogenate and serum samples were assayed by EIA. In vitro, progesterone- and 17β-oestradiol-induced VEGF production promoting cell proliferation and androgens are involved in the formation of vascular-like structures. In vivo, intratumoural testosterone concentrations were augmented and possibly associated with decreased metastatic rates, whereas elevated E1SO4 concentrations could promote tumour progression after antiangiogenic therapies. In conclusion, sex steroid hormones could regulate the production of angiogenic factors. The intratumoural measurement of sex steroids and growth factors may be useful to develop preventive and individualized therapeutic strategies.

Blocking Estrogen Synthesis Leads to Different Hormonal Responses in Canine and Human Triple Negative Inflammatory Breast Cancer

“…This subtype of breast cancer is characterized by a lack of estrogen receptor, progesterone receptor, and HER-2 and accounts for around 15% of breast cancers [ 9 ]. Despite its negativity to receptor expression, several TNBC tumors are able to synthetize steroid hormones locally that regulate tumor growth and progression, such as canine and human inflammatory breast cancer tumors [ 10 , 11 ]. Inflammatory breast cancer (IBC) in woman and inflammatory mammary cancer (IMC) in dogs represent a subtype of one of the worst mammary tumors, which accounts for 6% of diagnosed breast cancers and presents a poor survival rate [ 10 , 11 , 12 ].…”

“…Despite its negativity to receptor expression, several TNBC tumors are able to synthetize steroid hormones locally that regulate tumor growth and progression, such as canine and human inflammatory breast cancer tumors [ 10 , 11 ]. Inflammatory breast cancer (IBC) in woman and inflammatory mammary cancer (IMC) in dogs represent a subtype of one of the worst mammary tumors, which accounts for 6% of diagnosed breast cancers and presents a poor survival rate [ 10 , 11 , 12 ]. Comparative oncological studies had proposed the use of IMC tumors as a model for human disease research, due to its similarities in terms of clinical, molecular, and histological aspects in both species [ 10 , 11 ].…”

Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

“…In previous studies carried out by our group, possible local synthesis of some steroid hormones was indicated in normal and neoplastic mammary glands in canine mammary carcinoma, and more recently in hIBC and cIMC [1,26]. The formation of sex steroids in peripheral tissues in humans is well documented [39].…”

“…The greatest longitudinal diameter (length) and greatest transverse diameter (width) were measured. Tumor volume was estimated using the formula: volume = (length × width 2 )/2 [26]. When tumors reached a volume of 1.5 cm 3 (endpoint), blood samples were obtained intracardially and collected in heparin-coated tubes.…”

“…Determined steroid hormones in tumor homogenates were assayed using previously validated competitive enzyme-linked immunosorbent assay (ELISA), and an amplified ELISA was used for blood samples [26]. Briefly, 96-well flat-bottom medium-binding polystyrene microplates (Greiner Bio-One, Madrid, Spain) were coated with the appropriate purified antibody dilution overnight at 4 • C. Afterward, for competitive ELISA, plates were washed and standards and tumor homogenate samples were diluted in working solution (CWS) and analyzed in duplicate.…”