Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) contribute to the intimal hyperplasia in type 2 diabetes mellitus (T2DM) patients after percutaneous coronary intervention. We aimed to investigate the role of lncRNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) in VSMC proliferation and migration, as well as the underlying mechanism. T2DM model mice with carotid balloon injury were used in vivo and mouse aortic vascular smooth muscle cells (MOVAS) stimulated by insulin were used in vitro to assess the role of CDKN2B-AS1 in VSMC proliferation and migration following vascular injury in T2DM state. To investigate cell viability and migration, MTT assay and Transwell assay were conducted. To elucidate the underlying molecular mechanisms, the methylation-specific polymerase chain reaction, RNA immunoprecipitation, RNA-pull down, co-immunoprecipitation, and chromatin immunoprecipitation were performed. In vivo, CDKN2B-AS1 was up-regulated in common carotid artery tissues. In vitro, insulin treatment increased CDKN2B-AS1 level, enhanced MOVAS cell proliferation and migration, while the promoting effect was reversed by CDKN2B-AS1 knockdown. CDKN2B-AS1 forms a complex with enhancer of zeste homolog 2 (EZH2) and DNA methyltransferase (cytosine-5) 1 (DNMT1) to regulate smooth muscle 22 alpha (SM22α) methylation levels. In insulin-stimulated cells, SM22α knockdown abrogated the inhibitory effect of CDKN2B-AS1 knockdown on cell viability and migration. Injection of lentivirus-sh-CDKN2B-AS1 relieved intimal hyperplasia in T2DM mice with carotid balloon injury. Up-regulation of CDKN2B-AS1 induced by insulin promotes cell proliferation and migration by targeting SM22α through forming a complex with EZH2 and DNMT1, thereby aggravating the intimal hyperplasia after vascular injury in T2DM.

血管平滑肌细胞（VSMC）的过度增殖和迁移导致2型糖尿病（T2DM）患者经皮冠状动脉介入治疗后内膜增生。我们的目的是研究lncRNA细胞周期蛋白依赖性激酶抑制剂2B反义RNA 1（ CDKN2B-AS1 ）在VSMC增殖和迁移中的作用及其潜在机制。体内使用颈动脉球囊损伤的 T2DM 模型小鼠，体外使用胰岛素刺激的小鼠主动脉血管平滑肌细胞（MOVAS）来评估 CDKN2B-AS1 在 T2DM 状态血管损伤后 VSMC 增殖和迁移中的作用。为了研究细胞活力和迁移，进行了 MTT 测定和 Transwell 测定。为了阐明潜在的分子机制，进行了甲基化特异性聚合酶链式反应、RNA 免疫沉淀、RNA 下拉、免疫共沉淀和染色质免疫沉淀。在体内，CDKN2B-AS1在颈总动脉组织中表达上调。在体外，胰岛素治疗增加了CDKN2B-AS1水平，增强了MOVAS细胞的增殖和迁移，而CDKN2B-AS1敲低则逆转了促进作用。CDKN2B-AS1与 zeste 同源物增强子 2 (EZH2) 和 DNA 甲基转移酶 (胞嘧啶-5) 1 (DNMT1) 形成复合物，调节平滑肌 22 α ( SM22α ) 甲基化水平。在胰岛素刺激的细胞中，SM22α敲低消除了CDKN2B-AS1敲低对细胞活力和迁移的抑制作用。注射慢病毒-sh- CDKN2B-AS1可缓解颈动脉球囊损伤的T2DM小鼠的内膜增生。胰岛素诱导的CDKN2B-AS1上调，通过与EZH2和DNMT1形成复合物，靶向SM22α，促进细胞增殖和迁移，从而加重T2DM血管损伤后的内膜增生。