Differential expression of splicing variants of the human caldesmon gene (CALD1) in glioma neovascularization versus normal brain microvasculature.
Caldesmon is a cytoskeleton-associated protein which has not yet been related to neoplastic angiogenesis. In this study we investigated the expression of the caldesmon gene (CALD1) splicing variants and the protein expression level in glioma microvessels versus normal brain microvasculature. To exclude sources of splice variant expression from non-vascular components all possible cellular components present in control and glioma samples were pre-screened by laser-capture microdissection followed by RT-PCR before the cohort study. We discovered differential expression of the splicing variants of CALD1 in the tumor microvessels in contrast to normal brain microvasculature. Missplicing of exons 1, 1 + 4, and 1' + 4 of the gene is exclusively found in glioma microvessels. To exclude the possibility that this missplicing results from splice-site mutations, mutation scanning was performed by a coupled in vitro transcription/translation assay (IVTT). No premature stop mutations were traced by the IVTT. The transcriptional changes consequently resulted in up-regulation at the protein expression level. The up-regulated expression of caldesmon was coincident with the down-regulated expression of tight junction proteins (occludin and ZO-1). The results support the notion that missplicing of the CALD1 gene in glioma microvasculature is an independent epigenetic event regulated at the transcriptional level. The event coexists with tight junction (TJ) breakdown of the endothelial cells in glioma microvasculature. The data reveal a novel mechanism contributing to dysfunctionality of glioma neovascularization.
Zheng
Ping-Pin
PP
Department of Pathology, Internal Oncology, Neuro-Oncology and Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
Sieuwerts
Anieta M
AM
Luider
Theo M
TM
van der Weiden
M
M
Sillevis-Smitt
Peter A E
PA
Kros
Johan M
JM
D017398:Alternative Splicing / Q000235:genetics*; D001921:Brain / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D001932:Brain Neoplasms / Q000098:blood supply* / Q000235:genetics*; D002148:Calmodulin-Binding Proteins / Q000235:genetics*; D005091:Exons; D020869:Gene Expression Profiling; D014644:Genetic Variation / Q000235:genetics*; D005910:Glioma / Q000098:blood supply* / Q000235:genetics*; D006801:Humans; D009389:Neovascularization, Pathologic / Q000235:genetics* / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002148:Calmodulin-Binding Proteins