Yin Jian, Xiao Wei, Zhao Qingbin, Sun Jungang, Zhou Wenzheng, Zhao Wei
Department of Orthopedic, Xinjiang Uygur Autonomous Region People's Hospital, Urumqi, PR. China.
Immunopharmacol Immunotoxicol. 2022 Jun;44(3):421-428. doi: 10.1080/08923973.2022.2052895. Epub 2022 Mar 30.
Recent studies have demonstrated that micro RNAs (miRNAs) are involved in bone formation and bone cell differentiation, but the role of miR-582-3p in osteoporosis is unclear. We want to study the mechanism of miR-582-3p on osteogenic differentiation.
The expression of miR-582-3p and homeobox (Hox) A10 were analyzed by quantitative RT-PCR. The expression levels of HOXA10 protein were determined by Western blot. The target of HOXA10 was identified by bioinformatics and luciferase reporter gene assay.
The results showed that miR-582-3p was up-regulated in OP tissues and down-regulated in osteogenic differentiated C2C12 cells compared with that in the control group. Overexpression of miR-582-3p resulted in reduced expression levels of osteocalcin (OC), alkaline phosphatase (ALP), and collagen, type I, α1 (COL1A1). miR-582-3p had a potential binding site with HOXA10. Moreover, miR-582-3p inhibited the expression of HOXA10, and overexpression of HOXA10 reduced the effect of miR-582-3p on osteoblast markers. HOXA10 was the target gene of miR-582-3p, which could inhibit the expression of HOXA10. Furthermore, HOXA10 reduced the role of miR-582-3p in osteoblast markers. miR-582-3p inhibited the development of osteoporosis by regulating HOXA10 and osteoblast differentiation.
miR-582-3p may be a therapeutic target of osteoporosis treatment.
近期研究表明，微小RNA（miRNA）参与骨形成和骨细胞分化，但miR-582-3p在骨质疏松症中的作用尚不清楚。我们旨在研究miR-582-3p在成骨分化中的作用机制。
采用定量逆转录聚合酶链反应（RT-PCR）分析miR-582-3p和同源框（Hox）A10的表达。通过蛋白质印迹法检测HOXA10蛋白的表达水平。利用生物信息学和荧光素酶报告基因检测确定HOXA10的靶标。
结果显示，与对照组相比，miR-582-3p在骨质疏松症（OP）组织中上调，在成骨分化的C2C12细胞中下调。miR-582-3p过表达导致骨钙素（OC）、碱性磷酸酶（ALP）和I型胶原α1（COL1A1）的表达水平降低。miR-582-3p与HOXA10存在潜在结合位点。此外，miR-582-3p抑制HOXA10的表达，HOXA10过表达降低了miR-582-3p对成骨细胞标志物的影响。HOXA10是miR-582-3p的靶基因，miR-582-3p可抑制HOXA10的表达。此外，HOXA10减弱了miR-582-3p在成骨细胞标志物方面的作用。miR-582-3p通过调节HOXA10和成骨细胞分化来抑制骨质疏松症的发展。
miR-582-3p可能是骨质疏松症治疗的一个治疗靶点。
