摘要
目的:探讨右美托咪定对子宫内膜癌细胞的增殖、迁移和侵袭的影响和机制。方法:采用不同浓度(5,10,20,40μmol·L^(-1))的右美托咪定处理子宫内膜癌细胞Ishikawa,细胞计数试剂盒(CCK-8)检测细胞存活率,Transwell法检测细胞迁移和侵袭能力,并确定用药浓度。实时定量PCR(RT-qPCR)和蛋白质印迹法(Western blot)检测右美托咪定对Ishikawa细胞miR-219a-5p和细胞周期素依赖激酶2相关性Culin结构域1(CACUL1)表达的影响。采用上述方法检测抑制miR-219a-5p表达或过表达CACUL1对右美托咪定处理的Ishikawa细胞增殖、迁移和侵袭的影响。双荧光素酶报告基因实验和Western blot验证miR-219a-5p和CACUL1的靶向调控关系。结果:右美托咪定可抑制Ishikawa细胞增殖活力和迁移侵袭能力(P<0.05),且呈剂量依赖性。右美托咪定处理可促进miR-219a-5p表达,抑制CACUL1表达(P<0.05)。抑制miR-219a-5p表达或过表达CACUL1可逆转右美托咪定对Ishikawa细胞的增殖、迁移和侵袭的影响(P<0.05)。CACUL1是miR-219a-5p的靶基因,miR-219a-5p负性调控CACUL1表达。结论:右美托咪定通过调节miR-219a-5p/CACUL1轴抑制子宫内膜癌细胞的增殖、迁移和侵袭。
Objective: To investigate the effect of dexmedetomidine on the proliferation,migration and invasion of endometrial cancer cells and its mechanism. Methods: Ishikawa was treated with different concentrations(5,10,20 and 40 μmol · L-1) of dexmedetomidine,cell viability was detected by cell counting kit(CCK-8),and cell migration and invasion ability were detected by Transwellassay. Finally,the concentration was determined based on the above results. The effects of dexmedetomidine on the expressions of miR-219a-5p and cyclin-dependent kinase 2-associated Culin domain 1(CACUL1) in Ishikawa cells were detected by Realtime quantitative PCR(RT-QPCR) and Western blot. The effects of inhibiting miR-219a-5p or overexpressing CACUL1 on the proliferation,migration and invasion of Ishikawa cells treated with dexmedetomidine were detected by the above methods. The dual luciferase reporter gene assay and Western blot were used to verify the targeted and regulatory relationship between miR-219a-5p and CACUL1.Results: Dexmedetomidine inhibited the proliferation and migration of Ishikawa cells in a dose-dependent manner(P<0.05). Dexmedetomidine treatment promoted miR-219a-5p expression and inhibited CACUL1 expression(P<0.05). Inhibition of miR-219a-5p or over-expression of CACUL1 reversed the effect of dexmedetomidine on the proliferation,migration and invasion of Ishikawa cells(P <0.05). CACUL1 was a target gene of miR-219a-5p,and miR-219a-5p negatively regulated CACUL1 expression. Conclusion: Dexmedetomidine inhibits the proliferation,migration and invasion of endometrial cancer cells by regulating miR-219a-5p/CACUL1 axis.
作者
安燕晨
王翠花
魏伟
An Yanchen;Wang Cuihua;Wei Wei(Department of Gynaecology,Affiliated Hospital of Heza Medical College,Shandong Heze 274000,China)
出处
《中国药师》
CAS
2021年第2期237-241,246,共6页
China Pharmacist
