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2014 ; 34
(6
): 1249-59
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Identification and initial functional characterization of a human vascular
cell-enriched long noncoding RNA
#MMPMID24578380
Bell RD
; Long X
; Lin M
; Bergmann JH
; Nanda V
; Cowan SL
; Zhou Q
; Han Y
; Spector DL
; Zheng D
; Miano JM
Arterioscler Thromb Vasc Biol
2014[Jun]; 34
(6
): 1249-59
PMID24578380
show ga
OBJECTIVE: Long noncoding RNAs (lncRNAs) represent a rapidly growing class of RNA
genes with functions related primarily to transcriptional and
post-transcriptional control of gene expression. There is a paucity of
information about lncRNA expression and function in human vascular cells. Thus,
we set out to identify novel lncRNA genes in human vascular smooth muscle cells
and to gain insight into their role in the control of smooth muscle cell
phenotypes. APPROACH AND RESULTS: RNA sequencing (RNA-seq) of human coronary
artery smooth muscle cells revealed 31 unannotated lncRNAs, including a vascular
cell-enriched lncRNA (Smooth muscle and Endothelial cell-enriched
migration/differentiation-associated long NonCoding RNA [SENCR]). Strand-specific
reverse transcription polymerase chain reaction (PCR) and rapid amplification of
cDNA ends indicate that SENCR is transcribed antisense from the 5' end of the
FLI1 gene and exists as 2 splice variants. RNA fluorescence in situ hybridization
and biochemical fractionation studies demonstrate SENCR is a cytoplasmic lncRNA.
Consistent with this observation, knockdown studies reveal little to no
cis-acting effect of SENCR on FLI1 or neighboring gene expression. RNA-seq
experiments in smooth muscle cells after SENCR knockdown disclose decreased
expression of Myocardin and numerous smooth muscle contractile genes, whereas
several promigratory genes are increased. Reverse transcription PCR and Western
blotting experiments validate several differentially expressed genes after SENCR
knockdown. Loss-of-function studies in scratch wound and Boyden chamber assays
support SENCR as an inhibitor of smooth muscle cell migration. CONCLUSIONS: SENCR
is a new vascular cell-enriched, cytoplasmic lncRNA that seems to stabilize the
smooth muscle cell contractile phenotype.