Downregulation of desmuslin in primary vein incompetence

Abstract

Primary vein incompetence is one of the most common diseases of the peripheral veins, but its pathogenesis is unknown. These veins present obvious congenital defects, and examination of gene expression profiles of the incompetent vein specimens may provide important clues. The aim of this study was to screen for genes affecting the primary vein incompetence phenotype and test the differential expression of certain genes. We compared gene expression profiles of valvular areas from incompetent and normal great saphenous veins at the saphenofemoral junctions by fluorescent differential display reverse-transcription polymerase chain reaction (FDD RT-PCR). Differentially expressed complimentary DNAs (cDNAs) were confirmed by Northern blotting and semi-quantitative RT-PCR. Similarity of the cDNAs sequences to GenBank sequences was determined. Gene expression status was then determined by Western blot analysis and immunohistochemical techniques. There were >30 differentially expressed cDNA bands. Sequence analysis revealed that a cDNA fragment obviously downregulated in incompetent great saphenous vein was a portion of the messenger RNA (mRNA) encoding desmuslin, a newly discovered intermittent filament protein. Northern blotting and semi-quantitative RT-PCR analysis revealed a similar mRNA expression profile of the desmuslin gene in other samples. Western blotting and immunohistochemical techniques localized the desmuslin protein mainly in the cytoplasm of venous smooth muscle cells. The amount of desmuslin was greatly decreased in the smooth muscle cells of incompetent veins. The expression of many genes is altered in primary vein incompetence. Up- or downregulation of these genes may be involved in the pathogenesis of this disease. Desmuslin expression is downregulated in the abnormal veins. Its effect on the integrity of smooth muscle cells might be related to malformation of the vein wall. Further studies are needed to investigate other differentially expressed cDNAs and the exact role of desmuslin in this disease. Primary vein incompetence is a frequent and refractory disease of the peripheral veins. Exploring its pathogenesis may enhance our comprehension and management of this disease. We used reliable techniques to detect disease-related genes and confirmed downregulation of desmuslin in abnormal veins. Alteration of these genes might be used as disease markers or gene therapy targets.