Department of Orthopedics, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou, China.
Department of Anesthesiology, Weifang People's Hospital, Weifang, China.
FEBS Open Bio. 2020 Sep;10(9):1793-1801. doi: 10.1002/2211-5463.12929. Epub 2020 Jul 31.
Osteoporosis is a common disease that can seriously impair the physical and mental health of the elderly. However, current treatment is unsatisfactory due to the lack of effective therapeutic targets. Abnormal expression and involvement of microRNA-142 (miR-142) have been identified in many diseases, including bone-related diseases. Herein, we explored the effect of miR-142 on the viability, differentiation and apoptosis of the mouse preosteoblast cell line MC3T3-E1. We observed that the viability of MC3T3-E1 cells was significantly inhibited or promoted after transfection of miR-142 mimic or inhibitor, respectively. The apoptotic rate was dramatically increased by miR-142 mimic and decreased by inhibitor compared with the negative control group. Bcl-2 expression was down-regulated in the miR-142 mimic group and up-regulated in the miR-142 inhibitor group, whereas levels of cleaved caspase-3 and Bax were increased in the miR-142 mimic group and reduced in the miR-142 inhibitor group. Expression changes of Runx2 and Osteocalcin suggest that miR-142 inhibits the differentiation of osteoblast cells. Moreover, the luciferase reporter assay was used to verify that bone morphogenetic protein 2 (BMP2) is a target of miR-142. Overexpression of BMP2 repressed the proapoptotic effect of miR-142 mimic, whereas knockdown of BMP2 abolished the inhibitory effect of miR-142 inhibitor on the apoptosis of MC3T3-E1 cells. Furthermore, up-regulation or down-regulation of miR-142 dramatically decreased or increased the ratio of p-Smad1/5/Smad1 and p-Smad1/5/Smad5, respectively. Collectively, our results imply that miR-142 might influence the viability and differentiation of osteoblast cells by regulating BMP2 and BMP/Smad signaling.
骨质疏松症是一种常见疾病,会严重损害老年人的身心健康。然而,由于缺乏有效的治疗靶点,目前的治疗效果并不理想。在许多疾病中,包括骨相关疾病,已经发现 microRNA-142 (miR-142) 的异常表达和参与。在此,我们研究了 miR-142 对小鼠前成骨细胞系 MC3T3-E1 的活力、分化和凋亡的影响。我们观察到,miR-142 模拟物或抑制剂转染后,MC3T3-E1 细胞的活力明显受到抑制或促进。与阴性对照组相比,miR-142 模拟物组细胞凋亡率显著增加,而抑制剂组细胞凋亡率显著降低。miR-142 模拟物组 Bcl-2 表达下调,miR-142 抑制剂组 Bcl-2 表达上调,而 miR-142 模拟物组 cleaved caspase-3 和 Bax 水平升高,miR-142 抑制剂组 cleaved caspase-3 和 Bax 水平降低。Runx2 和 Osteocalcin 的表达变化表明 miR-142 抑制成骨细胞的分化。此外,还使用荧光素酶报告基因检测证实了骨形态发生蛋白 2 (BMP2) 是 miR-142 的靶基因。BMP2 的过表达抑制了 miR-142 模拟物的促凋亡作用,而 BMP2 的敲低则消除了 miR-142 抑制剂对 MC3T3-E1 细胞凋亡的抑制作用。此外,miR-142 的上调或下调分别显著降低或增加了 p-Smad1/5/Smad1 和 p-Smad1/5/Smad5 的比值。综上所述,我们的研究结果表明,miR-142 可能通过调节 BMP2 和 BMP/Smad 信号通路影响成骨细胞的活力和分化。