Zhang Hongfei, Li Jinglian, Shao Weiguang, Shen Naipeng
Department of Arthritis, Affiliated Hospital of Weifang Medical University, No. 2428, Yuhe Road, Kuiwen District, Weifang City, 261031, Shandong Province, China.
Weifang Medical University, NO.4948 Shengli East Street, Weifang City,, 261042, Shandong Province, China.
Clin Rheumatol. 2020 Nov;39(11):3473-3478. doi: 10.1007/s10067-020-05113-4. Epub 2020 May 10.
LncRNA CTBP1-AS2 has been reported to be involved in type 2 diabetes and cardiomyocyte hypertrophy, while its roles in other human diseases are unknown. Our preliminary deep sequencing analysis showed altered expression of CTBP1-AS2 in osteoarthritis (OA). In addition, CTBP1-AS2 was inversely correlated with miR-130a. This study was therefore carried out to investigate the interactions between CTBP1-AS2 and miR-130a in OA.
Synovial fluid was collected from 62 OA patients and 62 healthy controls. RT-qPCR was performed to determine the expression levels of CTBP1-AS2 and miR-130a in synovial fluid. Cell transfections were performed to investigate the interactions between CTBP1-AS2 and miR-130a. Methylation-specific PCR (MSP) was performed to assess the effects of CTBP1-AS2 on the methylation of miR-130a. Cell counting Kit-8 (CCK-8) assay was performed to evaluate the roles of CTBP1-AS2 and miR-130a in regulating proliferation of chondrocytes.
The results showed that CTBP1-AS2 was upregulated in OA and inversely correlated with miR-130a. In chondrocytes of OA patients, overexpression of CTBP1-AS2 led to increased methylation of miR-130a gene and downregulated expression of miR-130a, while overexpression of miR-130a did not affect the expression of CTBP1-AS2. In contrast, no interaction between CTBP1-AS2 and miR-130a was observed in chondrocytes from healthy adults. Analysis of chondrocyte proliferation showed that overexpression of miR-130a led to increased proliferation rate of chondrocytes extracted from OA patients. Overexpression of CTBP1-AS2 led to decreased proliferation rate of chondrocytes and reversed the effects of overexpressing miR-130a.
Therefore, CTBP1-AS2 is upregulated in OA and may increase the methylation of miR-130a gene to inhibit chondrocyte proliferation. Key Points • CTBP1-AS2 is overexpressed in OA and may downregulate miR-130a through methylation to suppress the proliferation of chondrocytes. • The interaction between CTBP1-AS2 and miR-130a is indirect and mediated by certain pathological mediators.
已有报道称长链非编码RNA CTBP1-AS2与2型糖尿病和心肌细胞肥大有关，但其在其他人类疾病中的作用尚不清楚。我们初步的深度测序分析显示骨关节炎（OA）中CTBP1-AS2的表达发生了改变。此外，CTBP1-AS2与miR-130a呈负相关。因此，本研究旨在探讨OA中CTBP1-AS2与miR-130a之间的相互作用。
收集62例OA患者和62例健康对照者的滑液。采用逆转录定量聚合酶链反应（RT-qPCR）检测滑液中CTBP1-AS2和miR-130a的表达水平。进行细胞转染以研究CTBP1-AS2与miR-130a之间的相互作用。采用甲基化特异性聚合酶链反应（MSP）评估CTBP1-AS2对miR-130a甲基化的影响。采用细胞计数试剂盒-8（CCK-8）法评估CTBP1-AS2和miR-130a在调节软骨细胞增殖中的作用。
结果显示，OA中CTBP1-AS2上调，且与miR-130a呈负相关。在OA患者的软骨细胞中，CTBP1-AS2的过表达导致miR-130a基因甲基化增加，miR-130a表达下调，而miR-130a的过表达不影响CTBP1-AS2的表达。相反，在健康成年人的软骨细胞中未观察到CTBP1-AS2与miR-130a之间的相互作用。软骨细胞增殖分析显示，miR-130a的过表达导致从OA患者中提取的软骨细胞增殖率增加。CTBP1-AS2的过表达导致软骨细胞增殖率降低，并逆转了miR-130a过表达的作用。
因此，OA中CTBP1-AS2上调，可能增加miR-130a基因的甲基化以抑制软骨细胞增殖。要点：•CTBP1-AS2在OA中过表达，可能通过甲基化下调miR-130a以抑制软骨细胞增殖。•CTBP1-AS2与miR-130a之间的相互作用是间接的，由某些病理介质介导。
