Ex vivo apoptotic and autophagic influence of an estradiol analogue on platelets

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Authors

Repsold, Lisa
Pretorius, Etheresia
Joubert, Annie M.

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BioMed Central

Abstract

BACKGROUND : Platelets are known contributors to the vascularization, metastasis and growth of tumors. Upon their interaction with cancer cells they are activated resulting in degranulation and release of constituents. Since the apoptotic- and autophagic effects of 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16) has been shown to occur in vitro and this compound was designed to bind to carbonic anhydrase II (CAII), the possible occurrence of these cell death mechanisms in platelets as circulatory components, is of importance. METHODS : Scanning electron microscopy was used to assess morphological changes in platelets after exposure to ESE-16. The possible apoptotic- and autophagic effect of ESE-16 in platelets was also determined by means of flow cytometry through measurement of Annexin V-FITC, caspase 3 activity, autophagy related protein 5 levels and light chain 3-I to light chain 3-II conversion. RESULTS : Scanning electron microscopy revealed no changes in ESE-16-treated platelets when compared to vehicletreated samples. Apoptosis detection by Annexin V-FITC and measurement of caspase 3 activity indicated that there was no increase in apoptosis when platelets were exposed to ESE-16. The incidence of autophagy by measurement of autophagy related protein 5 levels and light chain 3-I to light chain 3-II conversion showed that exposure to ESE-16 did not cause the incidence of autophagy in platelets. CONCLUSION : This is the first ex vivo study reporting on involvement of apoptosis- and autophagy-related targets in platelets after exposure to ESE-16, warranting further investigation in platelets of cancer patients.

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Platelets, Apoptosis, Autophagy, 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16), Carbonic anhydrase II (CAII), Cancer cells

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Citation

Repsold, L, Pretorius, E & Joubert, AM 2016, 'Ex vivo apoptotic and autophagic influence of an estradiol analogue on platelets', Experimental Hematology and Oncology, vol. 5, art. no. 18, pp. 1-10.