Chen Bing, Ji Fei, Wen Xinian, Jin Zhong
Department of Gastroenterology, The First Affiliated Hospital of Xinjiang Medical University Urumchi 830054, Xinjiang Uygur Autonomous Region, China.
Department of Gynecology, The First Affiliated Hospital of Xinjiang Medical University Urumchi 830054, Xinjiang Uygur Autonomous Region, China.
Int J Clin Exp Pathol. 2020 Nov 1;13(11):2806-2819. eCollection 2020.
Gastric cancer (GC) is one of the most common causes of cancer death. GSE83521 microarray analysis suggested that circular RNA circ_ASAP2 (hsa_circ_0008768) expression was increased in GC tissues. However, the molecular mechanism of circ_ASAP2 remains unknown.
Expression levels of circ_ASAP2, microRNA-770-5p (miR-770-5p), and the cyclin-dependent kinase 6 (CDK6) were detected by using real time PCR (RT-PCR). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and transwell assays were applied to explore cell viability, migration, and invasion, respectively. The interactions between miR-770-5p and circ_ASAP2 or CDK6 was predicted by using Starbase software, and then confirmed by luciferase reporter assay. Xenograft tumor model was also used to estimate the effect of circ_ASAP2 on tumor growth .
The expression levels of circ_ASAP2 and CDK6 were increased, and miR-770-5p level was decreased in GC tissues and cells. Furthermore, circ_ASAP2 knockdown inhibited cell viability, migration, and invasion of GC cells. Mechanically, circ_ASAP2 functioned as a sponge of miR-770-5p to regulate CDK6 expression, thereby boosting the progression of GC cells. Circ_ASAP2 silencing hindered the tumor growth of GC .
Circ_ASAP2 knockdown can repress the development of GC cells partly through regulating the miR-770-5p/CDK6 axis, suggesting an underlying circRNA-targeted therapy for GC treatment.
胃癌(GC)是癌症死亡的最常见原因之一。GSE83521基因芯片分析表明,环状RNA circ_ASAP2(hsa_circ_0008768)在GC组织中的表达增加。然而,circ_ASAP2的分子机制仍不清楚。
采用实时定量聚合酶链反应(RT-PCR)检测circ_ASAP2、微小RNA-770-5p(miR-770-5p)和细胞周期蛋白依赖性激酶6(CDK6)的表达水平。分别应用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐(MTT)法和Transwell实验探讨细胞活力、迁移和侵袭能力。使用Starbase软件预测miR-770-5p与circ_ASAP2或CDK6之间的相互作用,然后通过荧光素酶报告基因实验进行验证。还采用异种移植瘤模型评估circ_ASAP2对肿瘤生长的影响。
在GC组织和细胞中,circ_ASAP2和CDK6的表达水平升高,而miR-770-5p水平降低。此外,circ_ASAP2敲低抑制了GC细胞的活力、迁移和侵袭。机制上,circ_ASAP2作为miR-770-5p的海绵来调节CDK6的表达,从而促进GC细胞的进展。circ_ASAP2沉默阻碍了GC的肿瘤生长。
circ_ASAP2敲低可部分通过调节miR-770-5p/CDK6轴抑制GC细胞的发展,提示一种潜在的针对circRNA的GC治疗方法。