Mir-199a-3p Mediates Fluid Shear Stress-Induced Osteoblast Proliferation by Targeting CABLES-1
10.16156/j.1004-7220.2023.02.10
- VernacularTitle:miR-199a-3p 通过靶向 CABLES-1 调控流体剪切力介导的成骨细胞增殖
- Author:
Lifu WANG
1
,
2
,
3
;
Kun ZHANG
1
,
2
,
3
;
Qiong YI
1
,
2
,
3
;
Zhongcheng LIU
1
,
2
,
3
;
Xuening LIU
1
,
2
,
3
;
Bin GENG
1
,
2
,
3
;
Yayi XIA
1
,
2
,
3
Author Information
1. Department of Orthopedics, Lanzhou University Second Hospital
2. Orthopedic Clinical Medical Research Center of Gansu Province
3. Key Laboratory of Orthopedics of Gansu Province
- Publication Type:Journal Article
- Keywords:
fluid shear stress (FSS);
osteoblast;
cell proliferation;
miR-199a-3p
- From:
Journal of Medical Biomechanics
2023;38(2):E268-E275
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the role of miR-199a-3p in osteoblast proliferation induced by fluid shear stress (FSS) and the potential molecular mechanism. Methods Osteoblast MC3T3-E1 was treated with 1. 2 Pa FSS with time gradients of 0, 15, 30, 45, 60, 75 and 90 min, respectively. MC3T3-E1 cells were transfected with miR-199a-3p mimic or miR-199a-3p inhibitor. MC3T3-E1 cells were transfected with miR-199a-3p mimic and itsnegative control and then treated with 1. 2 Pa FSS for 45 min. The pc DNA NC, pc DNA-CABLES -1, si RNA NC and si RNA CABLES-1 were transfected into MC3T3-E1 cells. The pc DNA-CABLES-1 and mir-199a-3p mimic and SI NA-cables-1 and miR-199a-3p inhibitor were co-transfected, respectively. Cell activity was detected by CCK-8 assay. Real-time quantitative PCR (RT-qPCR) was used to detect expression levels of CABLES-1, miR-199a-3p, CDK 6, Cyclin D1 and PCNA. Luciferase reporting assay was used to detect targeting relationship between CABLES-1 and miR-199a-3p. Immunofluorescence was used to detect protein expression of CABLES-1.Western blot was used to detect protein expression of CABLES-1, CDK 6, PCNA and Cyclin D1. Results Mir- 199a-3p in MC3T3-E1 cells was significantly down-regulated by FSS. Over-expressed miR-199a-3p inhibitedosteoblast proliferation, and down-regulated miR-199a-3p expression promoted osteoblast proliferation. miR-199a- 3p could reverse the FSS-induced proliferation in osteoblasts. Dual luciferase assay showed that miR-199a-3p targeted to CABLES-1 and over-expressed miR-199a-3p inhibited expression of CBALES-1 protein. CABLES-1 could promote proliferation of osteoblasts. miR-199a-3p inhibited osteoblast proliferation induced by FSS through CABLES-1. Conclusions FSS-induced osteoblast proliferation can be realized by down-regulated miR-199a-3p expression via targeting CABLES-1. The findings in this study provide new direction for researches on mechanism of FSS-induced osteoblast proliferation, as well as new ideas for future research on clinical application of mechanical loading in the treatment of bone and joint diseases.
