Luo Junzhong, Han Jiuhui, Li Yazhou, Liu Yuchang
Department of Pediatric Orthopaedics, The Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
Oncol Lett. 2018 Jan;15(1):1319-1324. doi: 10.3892/ol.2017.7373. Epub 2017 Nov 8.
Legg-Calvé-Perthes disease (LCPD) commonly onsets in adolescents, and threatens their health. However, the potential mechanism underlying LCPD remains unclear. MicroRNA (miR)-206 and SRY-box 9 (SOX9) serve an important role in chondrocytes; however, their role in LCPD remains ambiguous. In the present study, whether miR-206 and SOX9 mediated cell apoptosis in dexamethasone (DEX)-induced LCPD was investigated. The chondrocytes of the LCPD and normal control group were isolated from clinical tissues. Reverse transcription-quantitative polymerase chain reaction was used to evaluate the expression of miR-206 and SOX9 mRNA. Western blotting was used to measure the protein level of SOX9. A combination of Annexin V-fluorescein isothiocyanate flow cytometry was used to assess cell apoptosis. The association between miR-206 and SOX9 was detected using a luciferase reporter assay. miR-206 was overexpressed while SOX9 was downregulated in chondrocytes treated with DEX obtained from patients with LCPD. miR-206 targeted SOX9 to regulate its expression. Overexpression of miR-206 promoted cell apoptosis in TC28, while it was reversed by SOX9 overexpression. TC28 cells pretreated with DEX significantly promoted cell apoptosis, while cells transfected with miR-206 inhibitor significantly reversed the effect; however, downregulated SOX9 abolished the effects of miR-206 inhibitor. SOX9 mediated by miR-206 possibly contributed to the pathogenesis of LCPD. The results of the present study suggest that miR-206 and SOX9 function as important therapeutic targets for the future of clinical therapy.
Legg-Calvé-Perthes病（LCPD）通常在青少年期发病，对他们的健康构成威胁。然而，LCPD潜在的发病机制仍不清楚。微小RNA（miR）-206和SRY盒9（SOX9）在软骨细胞中发挥重要作用；然而，它们在LCPD中的作用仍不明确。在本研究中，探讨了miR-206和SOX9是否介导地塞米松（DEX）诱导的LCPD中的细胞凋亡。从临床组织中分离出LCPD组和正常对照组的软骨细胞。采用逆转录-定量聚合酶链反应评估miR-206和SOX9 mRNA的表达。采用蛋白质印迹法检测SOX9蛋白水平。采用膜联蛋白V-异硫氰酸荧光素流式细胞术联合检测细胞凋亡。采用荧光素酶报告基因检测法检测miR-206与SOX9之间的关联。在LCPD患者来源的DEX处理的软骨细胞中，miR-206表达上调而SOX9表达下调。miR-206靶向SOX9以调节其表达。miR-206过表达促进TC28细胞凋亡，而SOX9过表达可逆转这一作用。预先用DEX处理的TC28细胞显著促进细胞凋亡，而转染miR-206抑制剂的细胞可显著逆转这一作用；然而，SOX9表达下调消除了miR-206抑制剂的作用。miR-2所介导的SOX9可能促成了LCPD的发病机制。本研究结果表明，miR-206和SOX9作为重要的治疗靶点，为未来的临床治疗指明了方向。
