MOLECULAR GENETIC RESPONSE OF ENDOTHELIAL CELLS TO DOXORUBICIN EXPOSURE: RESULTS OF IN VITRO AND IN VIVO STUDY

Highlights

• Doxorubicin is an anthracycline–type antibiotic used to treat certain forms of cancer, the use of which is associated with the development of a number of complications in the cardiovascular system.
• In vitro and in vivo experiments using primary endothelial cells of the coronary and internal thoracic arteries and normolipidemic Wistar rats have shown that exposure to doxorubicin leads to a change in the gene expression profile of endothelial cells, indicating a minor disruption of key links in the pathogenesis of endothelial dysfunction.

Abstract

Aim. To assess the expression profile of endothelial dysfunction marker genes in endothelial cells exposed to doxorubicin in in vitro and in vivo experiments. 

Method. The study included commercial cultures of primary human coronary and internal thoracic artery endothelial cells exposed to 2 μg/mL doxorubicin for 24 hours, and normolipidemic Wistar rats, which were injected with 2 μg/kg of body weight doxorubicin into the tail vein for four weeks. Endothelial dysfunction was assessed by measuring the expression of VCAM1, ICAM1, SELE, SELP, IL6, CXCL8, CCL2, CXCL1, MIF, VWF, SERPINE1, PLAU, PLAT, KLF2, KLF4, NFE2L2, NOS3, SNAI1, SNAI2, TWIST1, ZEB1, CDH5 and CDH2 genes in endothelial cell lysate and in endothelial monolayer washed from rat descending aorta using quantitative polymerase chain reaction.

Results. 2 μg/mL doxorubicin exposure is associated with upregulation of IL6, CXCL8 and CCL2 genes in both HCAEC and HITAEC, and downregulation of PECAM, MIF and NOS3 genes in HCAEC. The mRNA level of transcription factors ZEB1, VE-cadherin (CDH5) and CXCL1 was lower compared to the control in both cell lines. The assessment of gene expression in the endothelial lysate of aorta explanted from Wistar rats showed increased expression of the Ccl2 gene and the decreased expression of the Klf2, Plau, Nos3, Cdh2, Serpine and Vwf genes in the experimental group compared to the control.

Conclusion. In vitro and in vivo experiments using primary coronary and internal mammary artery endothelial cells and normolipidemic Wistar rats showed that doxorubicin exposure leads to some changes in the gene expression profile of endothelial cells, indicating minor disruption of key links in the pathogenesis of endothelial dysfunction.

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