Dicer knockdown induces fibronectin-1 expression in HEK293T cells via induction of Egr1.
BACKGROUND
Dicer is a multidomain ribonuclease III enzyme involved in the biogenesis of microRNAs (miRNAs) and small interfering RNAs (siRNAs); depletion of Dicer was found to impair the migration of endothelial cells.


METHODS
siRNA transfection, cell migration assay, real-time RT-PCR, chromatin immunoprecipitation, Western blotting, ELISA, caspase-3 activity assay, and annexin-V-FITC assay were utilized.


RESULTS
Knockdown of Dicer impairs the migratory capacity of HEK293T cells and induces fibronectin-1. The upregulation of fibronectin-1 is dependent on Egr1. Fibronectin-1/Dicer double-knockdown cells showed a marked increase in apoptosis compared with fibronectin-1 single knockdown cells.


CONCLUSIONS
Decreased Dicer expression induces fibronectin-1 expression via an Egr1-dependent manner.


CONCLUSIONS
Our data suggest that upregulation of fibronectin-1 protects Dicer knockdown HEK293T cells against apoptosis.
Tang
Kai-Fu
KF
Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, PR China.
Song
Guan-Bin
GB
Shi
Yi-Song
YS
Yuan
Lin
L
Li
Yong-Hua
YH
D017209:Apoptosis; D015153:Blotting, Western; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D002843:Chromatin / Q000235:genetics; D017931:DNA Primers; D051766:Early Growth Response Protein 1 / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D007668:Kidney / Q000196:embryology / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043244:Ribonuclease III / Q000235:genetics*; D015854:Up-Regulation
D002843:Chromatin; D017931:DNA Primers; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D005353:Fibronectins; D034741:RNA, Small Interfering; D043244:Ribonuclease III