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Overexpression of miR-126 inhibits the activation and migration of HSCs through targeting CRK.
- Source :
-
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology [Cell Physiol Biochem] 2014; Vol. 33 (1), pp. 97-106. Date of Electronic Publication: 2014 Jan 17. - Publication Year :
- 2014
-
Abstract
- Background & Aims: 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.<br />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.<br />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.<br />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.
- Subjects :
- Animals
Base Sequence
Cell Line
Cell Proliferation
Collagen Type I metabolism
Gene Expression Regulation
Male
MicroRNAs genetics
Molecular Sequence Data
Rats, Sprague-Dawley
Cell Movement
Hepatic Stellate Cells cytology
Hepatic Stellate Cells metabolism
MicroRNAs metabolism
Proto-Oncogene Proteins c-crk metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1421-9778
- Volume :
- 33
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
- Publication Type :
- Academic Journal
- Accession number :
- 24480980
- Full Text :
- https://doi.org/10.1159/000356653