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2022-02-10 - Colloque/Présentation - communication orale - Anglais - 1 page(s)

Scalia Alessandro , Kindt Nadège, Trelcat Anne , Nachtergael Amandine , Duez Pierre , Carlier Stéphane , "Development of 'home-made' oxLDL to study the interplay between atherosclerosis and cancer: characterization of their effects on head and neck cancer cells" in Belgian Society of Cardiology Meeting, Bruxelles, Belgique, 2022

  • Codes CREF : Cardiologie et circulation (DI3321), Chimie analytique (DI1314), Pharmacognosie (DI3410), Sciences pharmaceutiques (DI3400), Toxicologie pharmaceutique (DI3440)
  • Unités de recherche UMONS : Chimie thérapeutique et Pharmacognosie (M136)
  • Instituts UMONS : Institut des Sciences et Technologies de la Santé (Santé)

Abstract(s) :

(Anglais) Introduction Cardiovascular diseases (CVD) and cancers are the two main causes of death worldwide, sharing many comorbidities and risk factors. A pro-inflammatory state with neoangiogenesis and oxidative stress seems to be the cornerstone linking them, while initiation and progression of atherosclerotic plaque are mainly caused by oxidized low-density lipoproteins (oxLDL). Purpose We sought to produce in our laboratory well characterized oxLDL in order to control and standardize their production for further studies on cancer cells lines. We wanted to characterize our “home-made” oxLDL, comparing with commercially available oxLDL, and study their influence on the proliferation and migration of head and neck cancer cells (HNCC), both human papilloma virus (HPV)-positive and negative. Methods We applied a commercial purification kit based on serial precipitation and centrifugation to isolate LDL and VLDL from human plasma. LDL were separated from VLDL by gel permeation chromatography. LDL were then oxidized by incubation with 5 µM CuSO4 for 20 h, the reaction being stopped with 0.2mM EDTA. The quality of the oxidation was assessed by agarose gel electrophoresis to highlight the differential migrations of LDL and oxLDL in function of their electronegativity (*** V; ***°C; *** min). The expression of two oxLDL receptors, CD36 and LOX-1, was investigated by immunofluorescence on one HPV-positive HNCC (93VU-147T) and 3 HPV-negative HNCC (FaDu, Detroit 562 and UPCI-SCC-131) treated or not with oxLDL (30 µg/ml) over 48 h. The impact of oxLDL on cells migration was assessed over 48 h in Boyden chambers while their proliferation was estimated by microscopy upon staining with Crystal Violet. Results Electrophoresis showed oxLDL migrating, as expected, well farther than native LDL (25 mm against 11 mm, over a total electrophoresis pathway of *** mm) due to their increased electronegativity. Proliferation of HNCC increased with oxLDL exposition from 5 to 30 µg/mL, similarly in the 4 HNCC lines, independently of their HPV status. Beyond 30 µg/mL, the proliferation decreased, probably due to oxLDL toxicity. Both CD36 and LOX-1 expression increased in all HNCC after oxLDL exposition, in comparison with control. Migration decreased after oxLDL exposition, principally when oxLDL were in direct contact with HNCC. No difference was highlighted between "home-made" oxLDL and commercial oxLDL. Conclusions We successfully produced oxLDL comparable to commercially available ones to assess the involvement of oxLDL in cancer progression. Our results demonstrate increased HNCC proliferation under oxLDL exposition and a decrease on their migration. Further studies of the interplay between atherosclerosis and head and neck cancer cells, or others, are required to refine our understanding of the underlying mechanisms.