Effect of down-regulation of let-7c/g on triggering a double-negative feedback loop and promoting restenosis.
10.1097/CM9.0000000000002763
- Author:
Qian ZHANG
1
;
Xiaojun ZHOU
2
;
Xianzhi LI
3
;
Shuai YAO
1
;
Shan JIANG
1
;
Rui ZHANG
1
;
Zhiwei ZOU
1
;
Lin LIAO
2
;
Jianjun DONG
1
Author Information
1. Department of Endocrinology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China.
2. Department of Endocrinology and Metabology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong Key Laboratory of Rheumatic Disease and Translational Medicine, Shandong Institute of Nephrology, Jinan, Shandong 250012, China.
3. Department of Endocrinology and Metabology, Shandong Provincial Qianfoshan Hospital, Shandong University, Shandong Key Laboratory of Rheumatic Disease and Translational Medicine, Shandong Institute of Nephrology, Jinan, Shandong 250012, China.
- Publication Type:Journal Article
- MeSH:
Rats;
Animals;
Down-Regulation;
MicroRNAs/metabolism*;
Feedback;
Cell Proliferation/genetics*;
Atherosclerosis
- From:
Chinese Medical Journal
2023;136(20):2484-2495
- CountryChina
- Language:English
-
Abstract:
BACKGROUND:Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) are the main causes of restenosis (RS) in diabetic lower extremity arterial disease (LEAD). However, the relevant pathogenic mechanisms are poorly understood.
METHODS:In this study, we introduced a "two-step injury protocol" rat RS model, which started with the induction of atherosclerosis (AS) and was followed by percutaneous transluminal angioplasty (PTA). Hematoxylin-eosin (HE) staining and immunohistochemistry staining were used to verify the form of RS. Two-step transfection was performed, with the first transfection of Lin28a followed by a second transfection of let-7c and let-7g, to explore the possible mechanism by which Lin28a exerted effects. 5-ethynyl-2΄-deoxyuridine (EdU) and Transwell assay were performed to evaluate the ability of proliferation and migration of VSMCs. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to detect the expression of Lin28a protein and let-7 family members.
RESULTS:Using a combination of in vitro and in vivo experiments, we discovered that let-7c, let-7g, and microRNA98 (miR98) were downstream targets of Lin28a. More importantly, decreased expression of let-7c/let-7g increased Lin28a, leading to further inhibition of let-7c/let-7g. We also found an increased level of let-7d in the RS pathological condition, suggesting that it may function as a protective regulator of the Lin28a/let-7 loop by inhibiting the proliferation and migration of VSMCs.
CONCLUSION:These findings indicated the presence of a double-negative feedback loop consisting of Lin28a and let-7c/let-7g, which may be responsible for the vicious behavior of VSMCs in RS.
