TY - JOUR
T1 - Characterization of Circular RNAs in Vascular Smooth Muscle Cells with Vascular Calcification
AU - Ryu, Juhee
AU - Kwon, Duk Hwa
AU - Choe, Nakwon
AU - Shin, Sera
AU - Jeong, Geon
AU - Lim, Yeong Hwan
AU - Kim, Jaetaek
AU - Park, Woo Jin
AU - Kook, Hyun
AU - Kim, Young Kook
N1 - Publisher Copyright:
© 2019 The Authors
PY - 2020/3/6
Y1 - 2020/3/6
N2 - Circular RNAs (circRNAs) are generally formed by back splicing and are expressed in various cells. Vascular calcification (VC), a common complication of chronic kidney disease (CKD), is often associated with cardiovascular disease. The relationship between circRNAs and VC has not yet been studied. Inorganic phosphate (Pi) was used to treat rat vascular smooth muscle cells to induce VC. circRNAs were identified by analyzing RNA sequencing (RNA-seq) data, and their expression change during VC was validated. The selected circRNAs, including circSamd4a, circSmoc1-1, circMettl9, and circUxs1, were resistant to RNase R digestion and mostly localized in the cytoplasm. While silencing circSamd4a promoted VC, overexpressing it reduced VC in calcium assay and Alizarin red S (ARS) staining. In addition, microRNA (miRNA) microarray, luciferase reporter assay, and calcium assay suggested that circSamd4a could act as a miRNA suppressor. Our data show that circSamd4a has an anti-calcification role by functioning as a miRNA sponge. Moreover, mRNAs that can interact with miRNAs were predicted from RNA-seq and bioinformatics analysis, and the circSamd4a-miRNA-mRNA axis involved in VC was verified by luciferase reporter assay and calcium assay. Since circSamd4a is conserved in humans, it can serve as a novel therapeutic target in resolving VC.
AB - Circular RNAs (circRNAs) are generally formed by back splicing and are expressed in various cells. Vascular calcification (VC), a common complication of chronic kidney disease (CKD), is often associated with cardiovascular disease. The relationship between circRNAs and VC has not yet been studied. Inorganic phosphate (Pi) was used to treat rat vascular smooth muscle cells to induce VC. circRNAs were identified by analyzing RNA sequencing (RNA-seq) data, and their expression change during VC was validated. The selected circRNAs, including circSamd4a, circSmoc1-1, circMettl9, and circUxs1, were resistant to RNase R digestion and mostly localized in the cytoplasm. While silencing circSamd4a promoted VC, overexpressing it reduced VC in calcium assay and Alizarin red S (ARS) staining. In addition, microRNA (miRNA) microarray, luciferase reporter assay, and calcium assay suggested that circSamd4a could act as a miRNA suppressor. Our data show that circSamd4a has an anti-calcification role by functioning as a miRNA sponge. Moreover, mRNAs that can interact with miRNAs were predicted from RNA-seq and bioinformatics analysis, and the circSamd4a-miRNA-mRNA axis involved in VC was verified by luciferase reporter assay and calcium assay. Since circSamd4a is conserved in humans, it can serve as a novel therapeutic target in resolving VC.
KW - circSamd4a
KW - circular RNA
KW - microRNA
KW - vascular calcification
KW - vascular smooth muscle cells
UR - http://www.scopus.com/inward/record.url?scp=85075735638&partnerID=8YFLogxK
U2 - 10.1016/j.omtn.2019.11.001
DO - 10.1016/j.omtn.2019.11.001
M3 - Article
AN - SCOPUS:85075735638
SN - 2162-2531
VL - 19
SP - 31
EP - 41
JO - Molecular Therapy Nucleic Acids
JF - Molecular Therapy Nucleic Acids
ER -
