Wang Yulong, Zhai Shuiting, Xing Jianwu, He Yingkun, Li Tianxiao
Department of Intervention Center, Zhengzhou University People's Hospital, Henan Provincial People's Hospital, Zhengzhou.
Department of Intervention Section, Second Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan, China.
Anticancer Drugs. 2022 Mar 1;33(3):225-234. doi: 10.1097/CAD.0000000000001090.
One of the causes of abdominal aortic aneurysm (AAA) is the apoptosis of vascular smooth muscle cells. Many long noncoding RNA (lncRNAs) have been implicated in AAA formation. However, the mechanism of growth arrest-specific 5 (GAS5) in AAA formation is not yet clear. The expression levels of GAS5, microRNA-185-5p (miR-185-5p) and adenylate cyclase 7 (ADCY7) were determined by quantitative real-time PCR. Angiotensin II (ANGII) was used to induce AAA cell models. Cell viability was detected by MTT assay, and cell apoptosis was assessed by flow cytometry. Western blot analysis was used to test the protein expression levels. Besides, a dual-luciferase reporter assay was used to identify the mechanism of GAS5. GAS5 was upregulated in AAA tissues and ANGII-induced human aortic smooth muscle cells (HASMCs). GAS5 overexpression inhibited proliferation and promoted apoptosis and inflammatory response in ANGII-induced HASMCs, while its knockdown had the opposite effects. MiR-185-5p could be absorbed by GAS5, and its inhibitor could invert the effects of GAS5 silencing on proliferation, apoptosis and inflammatory response in ANGII-induced HASMCs. ADCY7 was a target of miR-185-5p. ADCY7 knockdown increased proliferation, while decreased apoptosis and inflammatory response in ANGII-induced HASMCs. Also, overexpressed ADCY7 reversed the effect of miR-185-5p overexpression on proliferation, apoptosis and inflammatory response in ANGII-induced HASMCs. GAS5 positively regulated the ADCY7 expression to inhibit the activity of the AKT signaling pathway by sponging miR-185-5p. LncRNA GAS5 contributed to AAA formation through regulating HASMCs proliferation, apoptosis and inflammatory response, which might provide new ideas for the treatment of AAA.
腹主动脉瘤（AAA）的病因之一是血管平滑肌细胞的凋亡。许多长链非编码RNA（lncRNAs）与AAA的形成有关。然而，生长停滞特异性5（GAS5）在AAA形成中的机制尚不清楚。通过定量实时PCR测定GAS5、微小RNA-185-5p（miR-185-5p）和腺苷酸环化酶7（ADCY7）的表达水平。使用血管紧张素II（ANGII）诱导AAA细胞模型。通过MTT法检测细胞活力，通过流式细胞术评估细胞凋亡。蛋白质印迹分析用于检测蛋白质表达水平。此外，使用双荧光素酶报告基因测定法来确定GAS5的机制。GAS5在AAA组织和ANGII诱导的人主动脉平滑肌细胞（HASMCs）中上调。GAS5过表达抑制ANGII诱导的HASMCs的增殖并促进其凋亡和炎症反应，而其敲低则产生相反的效果。MiR-185-5p可以被GAS5吸附，其抑制剂可以逆转GAS5沉默对ANGII诱导的HASMCs的增殖、凋亡和炎症反应的影响。ADCY7是miR-185-5p的靶标。ADCY7敲低增加了ANGII诱导的HASMCs的增殖，同时降低了其凋亡和炎症反应。此外，过表达的ADCY7逆转了miR-185-5p过表达对ANGII诱导的HASMCs的增殖、凋亡和炎症反应的影响。GAS5通过海绵化miR-185-5p正向调节ADCY7的表达，从而抑制AKT信号通路的活性。lncRNA GAS5通过调节HASMCs的增殖、凋亡和炎症反应促进AAA的形成，这可能为AAA的治疗提供新思路。
