MicroRNA-19a mediates the suppressive effect of laminar flow on cyclin D1 expression in human umbilical vein endothelial cells.
Endothelial cells (ECs) respond to changes in mechanical forces, leading to the modulation of signaling networks and cell function; an example is the inhibition of EC proliferation by steady laminar flow. MicroRNAs (miRs) are short noncoding 20-22 nucleotide RNAs that negatively regulate the expression of target genes at the posttranscriptional level. This study demonstrates that miRs are involved in the flow regulation of gene expression in ECs. With the use of microRNA chip array, we found that laminar shear stress (12 dyn/cm(2), 12 h) regulated the EC expression of many miRs, including miR-19a. We further showed that stable transfection of miR-19a significantly decreased the expression of a reporter gene controlled by a conserved 3'-untranslated region of the cyclinD1 gene and also the protein level of cyclin D1, leading to an arrest of cell cycle at G1/S transition. Laminar flow suppressed cyclin D1 protein level, and this suppressive effect was diminished when the endogenous miR-19a was inhibited. In conclusion, we demonstrated that miR-19a plays an important role in the flow regulation of cyclin D1 expression. These results revealed a mechanism by which mechanical forces modulate endothelial gene expression.
Qin
Xiaomei
X
Key Laboratory of Molecular Cardiovascular Science, Peking University Health Science Center, Beijing 100191, China.
Wang
Xiaohong
X
Wang
Ying
Y
Tang
Zhihui
Z
Cui
Qinghua
Q
Xi
Jianzhong
J
Li
Yi-Shuan J
YS
Chien
Shu
S
Wang
Nanping
N
D020029:Base Pairing; D001483:Base Sequence; D001696:Biomechanical Phenomena; D015153:Blotting, Western; D019938:Cyclin D1 / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism*; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D046228:Microarray Analysis; D008969:Molecular Sequence Data; D012039:Regional Blood Flow / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014471:Umbilical Veins / Q000166:cytology* / Q000502:physiology
D017931:DNA Primers; C549214:MIRN19A microRNA, human; D035683:MicroRNAs; D019938:Cyclin D1