Canfeng Li1, Qinshen Hu1, Zhuo Chen2, Bin Shen3, Jing Yang1, Pengde Kang1, Zongke Zhou1, Fuxing Pei1. 1. Department of Orthopedics, West China Hospital, Sichuan University, Chengdu, Sichuan, China. 2. Department of Transformational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, Sichuan, China. 3. Department of Orthopedics, West China Hospital, Sichuan University, Chengdu, Sichuan, China. Electronic address: shenbin71@hotmail.com.
Abstract
BACKGROUND: This study aimed to investigate the expression levels and relationship of bone morphogenetic proteins (BMPs) signaling molecules and microRNA-140 (miR-140) in human osteoarthritis (OA) chondrocytes. MATERIALS AND METHODS: Different stage chondrocytes (normal cartilage, mid-stage OA and advanced-stage OA) were isolated from cartilage samples according to Kellgren and Lawrence criteria. The effect of miR-140 on BMPs signaling was evaluated by transfecting miR-140 mimic or inhibitor into chondrocytes. The expression of responsive genes was measured using real-time polymerase chain reaction and Western blotting analysis. RESULTS: There was a significant reduction in miR-140 and SOX9 expression in OA groups compared to the normal group, and there was a further reduction in the severe OA group compared to the moderate OA group. Compared with the normal group, the expression of ALK1, SMAD1, COL10A1 and MMP3 was higher in the OA groups, whereas the expression of COL2A1 was lower in the OA groups. In the moderate OA group, transfection with miR-140 mimic increased SMAD1, SOX9 and COL2A1 expression, but decreased COL10A1 expression. However, there was an opposite effect after transfecting miR-140 inhibitor with decreased SMAD1, SOX9 and COL2A1 expression, and increased COL10A1 expression. Interestingly, the biological effect of transfecting miR-140 mimic or inhibitor was similar in the severe OA group. SMAD1 and COL2A1 protein production followed the same pattern as their expression profile. CONCLUSIONS: miR-140 suppresses chondrocytes hypertrophy by controlling the BMPs signaling pathway, which highlights the importance of miR-140 in the maintenance of chondrocyte homeostasis and opens up novel avenues in OA therapeutic strategies.
BACKGROUND: This study aimed to investigate the expression levels and relationship of bone morphogenetic proteins (BMPs) signaling molecules and microRNA-140 (miR-140) in humanosteoarthritis (OA) chondrocytes. MATERIALS AND METHODS: Different stage chondrocytes (normal cartilage, mid-stage OA and advanced-stage OA) were isolated from cartilage samples according to Kellgren and Lawrence criteria. The effect of miR-140 on BMPs signaling was evaluated by transfecting miR-140 mimic or inhibitor into chondrocytes. The expression of responsive genes was measured using real-time polymerase chain reaction and Western blotting analysis. RESULTS: There was a significant reduction in miR-140 and SOX9 expression in OA groups compared to the normal group, and there was a further reduction in the severe OA group compared to the moderate OA group. Compared with the normal group, the expression of ALK1, SMAD1, COL10A1 and MMP3 was higher in the OA groups, whereas the expression of COL2A1 was lower in the OA groups. In the moderate OA group, transfection with miR-140 mimic increased SMAD1, SOX9 and COL2A1 expression, but decreased COL10A1 expression. However, there was an opposite effect after transfecting miR-140 inhibitor with decreased SMAD1, SOX9 and COL2A1 expression, and increased COL10A1 expression. Interestingly, the biological effect of transfecting miR-140 mimic or inhibitor was similar in the severe OA group. SMAD1 and COL2A1 protein production followed the same pattern as their expression profile. CONCLUSIONS:miR-140 suppresses chondrocytes hypertrophy by controlling the BMPs signaling pathway, which highlights the importance of miR-140 in the maintenance of chondrocyte homeostasis and opens up novel avenues in OA therapeutic strategies.
Authors: Eliana Lara-Barba; María Jesús Araya; Charlotte Nicole Hill; Felipe A Bustamante-Barrientos; Alexander Ortloff; Cynthia García; Felipe Galvez-Jiron; Carolina Pradenas; Noymar Luque-Campos; Gabriela Maita; Roberto Elizondo-Vega; Farida Djouad; Ana María Vega-Letter; Patricia Luz-Crawford Journal: Front Immunol Date: 2021-11-01 Impact factor: 7.561