Mechanisms of skin toxicity of paclitaxel: An in vitro preclinical assessment.

Abstract

e15511 Background: Paclitaxel skin toxicity is a frequent side effect extensively evaluated in the clinical setting. However little is known about the preclinical mechanisms that lead to this toxicity. The endpoint of this study was to analyse the cutaneous mechanisms that drive paclitaxel toxicity in a preclinical model. Methods: Primary human keratinocytes were co-cultured with human dermal fibroblast in collagen gel under air-liquid interface conditions to generate a multilayered 3D epidermis. Paclitaxel was added to 3D epidermis at 0.3 µM, 3 µM and 30 µM and total RNA and protein was extracted after 24h of incubation. Markers of cell senescence (p21 and p53), anti-apoptotic mediators (Bcl-2), skin elasticity (tropoelastin ELN, collagen type I and fibronectin), hidratation (aquaporin 3 AQP3), oxidative stress (NOX4), antioxidant (SOD1, Nrf2), and angiogenic markers (VEGFR, eNOS) were evaluated by RT-PCR and western blot. NfKB phosphorylation was measured by western blot and inflammatory citokines IL1α, IL-6 and IL-8 were measured in cultured supernatants by ELISA. Human primary melanocytes were cultured and stimulated with paclitaxel to measure melanogenesis through the expression of of TYR, TYRP1 and DCT genes. The effect of paclitaxel on skin endothelial cell angiogenesis was measured by endothelial tube formation. Results: In human 3D keratinocytesPaclitaxel inhibited the expression of Bcl-2, and increased the expression of p53 and p21. The angiogenic markers VEGF and eNOS were decreased. The expression of oxidative stress marker NOX4 was increased and the anti-oxidant mediators Nrf2 and SOD1 were decreased. Markers of elasticity, collagen type I, fibronectin and FN1 were decreased as occurs with AQP3 hidratation marker. Paclitaxel increased the phosphorylation of NfkB and elevated the secretion of IL1α, IL-6 and IL-8 cytokines. In skin endothelial cells, paclitaxel reduced the endothelial tube formation. In melanocytes, paclitaxel increased skin pigmentation represented by the increase of RNA expression of TYR, TYRP1 and DCT genes. Conclusions: This preclinical 3D model showed that paclitaxel impacts on the expression of proteins related with angiogenics, elasticity and senescence in human kerantinocytes. Moreover higher doses of paclitaxel increased inflammation parameters and confirmed phototoxic and anti-angiogenenic effects. This preclinical model could be a valuable tool to assess skin toxicity of new antineoplastic agents.

Natural Taxanes: From Plant Composition to Human Pharmacology and Toxicity

“…It is directly cytotoxic to keratinocytes, causing allergic and hypersensitive reactions of type I and IV, which affect, among other parts, the skin [ 136 ]. Another mechanism of toxicity is inhibition of Bcl-2 expression, increase in p53 and p21 expression, decrease in vascular endothelial growth factor (VEGF), increase in markers of oxidative stress and proinflammatory markers, decrease in markers of elasticity and increased pigment production in melanocytes [ 137 ]. The most common taxane-induced skin symptoms typical of a hypersensitivity reaction are rash and pruritus, which are not serious.…”

Paclitaxel Induces Epidermal Molecular Changes and Produces Subclinical Alterations in the Skin of Gynecological Cancer Patients

“…Further, studies in vitro have described that paclitaxel induces a cytotoxic response in transformed HaCat keratinocytes [ 18 ] and produces epithelial damage in zebrafish models [ 19 , 20 ]. Further, our previous results showed that paclitaxel impacts on the expression of proteins related to angiogenesis, elasticity, inflammation, and senescence in human keratinocytes [ 21 ].…”