title: Silencing of PKC-α, TRPC1 or NF-κB expression attenuates cisplatin-induced ICAM-1 expression and endothelial dysfunction.
abstract: Platinum-based chemotherapy has been associated with increased long-term cardiovascular events. Also noteworthy is the accumulating awareness of early vascular toxicity occurring at the time of chemotherapy or immediately thereafter. The objective of the study was to delineate the molecular mechanisms associated with the early vascular toxicity and test the molecular silencing approach towards attenuating the endothelial dysfunction during platinum-based chemotherapy. Human umbilical vein endothelial cells (HUVECs) were treated with varying concentrations of cisplatin (1.0-10.0μg/ml) or vehicle control (0.1% dimethyl sulfoxide) for monitoring the changes in Intercellular adhesion molecule-1 (ICAM-1) mRNA and protein expression viz. a viz. altered activation of protein kinase C (PKC) isoforms, transient receptor potential channel (TRPC) 1 expression, Nuclear factor 'kappa-light-chain-enhancer' of activated B-cells (NF-κB), Store Operated Ca(2+) Entry (SOCE) in cisplatin-induced endothelial permeability and adherence of the activated endothelial cells to human monocyte-like U937 cells. Silencing of either PKC-α, TRPC1 or p65 subunit of NF-κB, all resulted in significant alleviation of cisplatin-induced endothelial dysfunction. At concentrations ≥8μg/ml, cisplatin induced a significant increase in the expression of ICAM-1 mRNA as well as protein. This was mediated by changes in PKC-α membrane translocation, NF-κB activation, increased expression as well as phosphorylation of TRPC1 and enhanced SOCE, leading to hyperpermeability and leakage of albumin. Increased adherence of U937 monocytes to cisplatin-activated endothelial cells was evident. Cisplatin challenge activates PKC-α, which in turn phosphorylated TRPC1 resulting in enhanced Ca(2+) entry. Increased Ca(2+) flux is required for activation of NF-κB and ICAM-1 expression. Enhanced ICAM-1 expression promotes monocyte binding to endothelial cells and increased endothelial hyperpermeability.
authors: Department of Molecular Biology, Institute of Genetics & Hospital for Genetic Diseases, Begumpet, Osmania University, Hyderabad 500016, Telangana, India.
authors: Vijaya Lakshmi
authors: VL
authors: Bodiga
authors: Department of Biochemistry, Kakatiya University, Vidyaranyapuri, Warangal 506009, Telangana, India.
authors: Madhukar Rao
authors: MR
authors: Kudle
authors: Department of Biochemistry, Kakatiya University, Vidyaranyapuri, Warangal 506009, Telangana, India.
authors: Sreedhar
authors: S
authors: Bodiga
mesh_terms: D002118:Calcium / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D020868:Gene Silencing; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D051571:Protein Kinase C-alpha / Q000235:genetics / Q000378:metabolism*; D050052:TRPC Cation Channels / Q000235:genetics / Q000378:metabolism*; D020298:U937 Cells
chemical_list: D016328:NF-kappa B; D050052:TRPC Cation Channels; C096351:transient receptor potential cation channel, subfamily C, member 1; D018799:Intercellular Adhesion Molecule-1; D051571:Protein Kinase C-alpha; D002945:Cisplatin; D002118:Calcium
keywords: Cisplatin; Endothelium; ICAM; NF-κB; PKC; SOCE; TRPC