Effect of LIMK2 RNAi on reorganization of the actin cytoskeleton in osteoblasts induced by fluid shear stress.
The biomechanical characteristics of bone tissue and its cells under mechanical stress are significant for bone biomechanics research, but the mechanism of mechanotransduction is still unknown. It has been established that the actin cytoskeleton of osteoblasts plays an important role in this process. However, the structure of the actin cytoskeleton is reorganized when loaded with mechanical stress, which results in changes in cell stiffness. These phenomena suggest that an actin-cytoskeleton-induced feedback regulation mechanism may be involved in the mechanotransduction of osteoblasts, but this has not yet been proven. The aim of this study was to explore the role of LIMK2 in the reorganization of the actin cytoskeleton induced by fluid shear stress in osteoblasts by using RNA interference. Balb/c mouse primary osteoblasts were divided into four groups. Cells in Groups 1 and 3 were transfected with negative control RNA, while cells in Groups 2 and 4 were transfected with a specific siRNA designed to silence the LIMK2 gene. Twenty-four hours after transfection, cells in Groups 1 and 2 were loaded with fluid shear stress at 12 dyne/cm2 while cells in Groups 3 and 4 were not. Compared with Group 1, the mean fluorescence density of the actin cytoskeleton in the other three groups was 28.9%, 45.7%, and 33.0%, respectively. These results indicate that LIMK2 plays an important role in the reorganization of the actin cytoskeleton induced by fluid shear stress.
Fu
Qiang
Q
Department of Prosthodontics, Guanghua School of Stomatology, Sun Yat-Sen University, 56#, Lingyuan Xi Road, Guangzhou, Guangdong 510056, PR China. fuqiangds@yahoo.com.cn
Wu
Changjing
C
Shen
Yun
Y
Zheng
Shucan
S
Chen
Rui
R
D000199:Actins / Q000502:physiology*; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure*; D054461:Lim Kinases / Q000235:genetics / Q000378:metabolism*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D044085:Microfluidics / Q000379:methods; D010006:Osteoblasts / Q000502:physiology* / Q000648:ultrastructure*; D034622:RNA Interference; D033081:Shear Strength; D013314:Stress, Mechanical
D000199:Actins; D054461:Lim Kinases; C095325:Limk2 protein, mouse