Overexpression of miR-126 inhibits the activation and migration of HSCs through targeting CRK.
OBJECTIVE
MicroRNAs (miRNAs) have been shown to play essential roles in HSCs activation which contributes to hepatic fibrosis. Our previous miRNA microarray results suggested that miR-126 might be decreased during HSCs activation as other studies. The aim of this study is to investigate the role of miR-126 during HSCs activation.


METHODS
In this study, the expression of miR-126 during HSCs activation was measured and confirmed by qRT-PCR. Then, miR-126 expression was restored by transfection of lentivirus vector encoding miR-126. Futhermore, cell proliferation was assayed by the cell counting kit-8 (CCK-8), cell migration was assayed by transwell assay, and the markers of activation of HSCs, α-SMA and collagen type I, were assayed by qRT-PCR, Western Blotting, Immunostaining and ELISA. Luciferase reporter assay was used to find the target of miR-126, and Western Blotting and Immunostaining was used to validate the target of miR-126. Then, the expression and the role of the target of miR-126 during HSCs activation was further assessed.


RESULTS
The expression of miR-126 was confirmed to be significantly decreased during HSCs activation. Overexpression of miR-126 significantly inhibited HSCs migration but did not affect HSCs proliferation. The expression of α-SMA and collagen type I were both obviously decreased by miR-126 restoration. CRK was found to be the target of miR-126 and overexpression of miR-126 significantly inhibited CRK expression. And it was found that overexpression of CRK also significantly decreased miR-126 expression and promoted HSCs activation.


CONCLUSIONS
Our study showed that overexpression of miR-126 significantly inhibited the activation and migration of HSCs through targeting CRK which can also decrease miR-126 expression and promote HSCs activation.
Department of Gastroenterology, Changzheng Hospital, Second Military Medical University, Shanghai, China.
Xu-Hua
XH
Gong
Chao
C
Chen
Peng
P
Hou
Song-Cheng
SC
Zhu
Chao-Qun
CQ
Wu
Chen-Lin
CL
Song
Wu
W
Ni
Jiang-Feng
JF
Hu
Ding-Kang
DK
Yao
Jiu-Hong
JH
Kang
Liang
L
Zhu
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D024042:Collagen Type I / Q000378:metabolism; D005786:Gene Expression Regulation; D055166:Hepatic Stellate Cells / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D050737:Proto-Oncogene Proteins c-crk / Q000378:metabolism*; D017207:Rats, Sprague-Dawley
D024042:Collagen Type I; C493872:Crk protein, rat; C571458:MIRN126 microRNA, rat; D035683:MicroRNAs; D050737:Proto-Oncogene Proteins c-crk