MSC-AS1对宫颈癌细胞生物学行为的影响及其机制

Effects of MSC-AS1 on the biological behavior of cervical cancer cells and the mechanism

目的探索长链非编码RNA(lncRNA)MSC-AS1在宫颈癌细胞生物行为中的作用和机制。方法qRT-PCR和Western blotting检测MSC-AS1、miR-520d-3p和三磷酸腺苷酶家族蛋白2(ATAD2)在宫颈癌组织和SiHa细胞中的表达。SiHa细胞分为si-NC组、si-MSC-AS1组、vector组、pc3.1-MSC-AS1组、miR-NC组、miR-520d-3p组、si-NC+anti-miR-NC组、si-MSC-AS1+anti-miR-NC组、si-MSC-AS1+anti-miR-520d-3p组。流式细胞术、Transwell实验分别检测细胞凋亡、迁移和侵袭水平的变化。软件预测结合双荧光素酶活性实验分析MSC-AS1、miR-520d-3p、ATAD2之间的靶向调控关系。结果与癌旁组织比较,宫颈癌组织MSC-AS1、ATAD2表达上调,miR-520d-3p表达下调(P<0.05)。SiHa细胞凋亡率、miR-520d-3p表达水平在沉默MSC-AS1后si-MSC-AS1组高于si-NC组,过表达MSC-AS1后pc3.1-MSC-AS1组低于vector组;沉默MSC-AS1同时抑制miR-520d-3p后si-NC+anti-miR-NC组<si-MSC-AS1+anti-miR-520d-3组<si-MSC-AS1+anti-miR-NC组(P<0.05)。MSC-AS1靶向miR-520d-3p,且miR-520d-3p靶向ATAD2,miR-520d-3p组SiHa细胞凋亡率高于miR-NC组(P<0.05)。SiHa细胞ATAD2蛋白的表达、迁移和侵袭细胞数si-MSC-AS1组低于si-NC组,pc3.1-MSC-AS1组高于vector组,miR-520d-3p组低于miR-NC组,si-NC+anti-miR-NC组>si-MSC-AS1+anti-miR-520d-3p组>si-MSC-AS1+anti-miR-NC组(P<0.05)。结论沉默MSC-AS1可能通过miR-520d-3p靶向ATAD2,抑制宫颈癌细胞的生长和转移,并促进凋亡。

Aim To explore the role and mechanism of long non-coding RNA(lncRNA)MSC-AS1 in the biological behavior of cervical cancer cells.Methods qRT-PCR and Western blotting were used to detect the expression of MSC-AS1,miR-520d-3p and ATAD2 in cervical cancer tissues and SiHa cells.SiHa cells were divided into si-NC group,si-MSC-AS1 group,vector group,pc3.1-MSC-AS1 group,miR-NC group,miR-520d-3p group,si-NC+anti-miR-NC group,si-MSC-AS1+anti-miR-NC group,and si-MSC-AS1+anti-miR-520d-3p group.Flow cytometry,and Transwell experiment were used to evaluate the changes of cell apoptosis,and migration and invasion levels,respectively.The targeted regulatory relationships among MSC-ASI,miR-520d-3p,ATAD2 were analyzed by combining starbase software and targetscan software with luciferase activity experiments.Results Compared with adjacent tissues,the expression of MSC-AS1 and ATAD2 protein in cervical cancer tissues were upregulated,while miR-520d-3p was downregulated(P<0.05).After silencing MSC-AS1,the expression of miR-520d-3p and apoptosis rate of SiHa cells were higher in the si-MSC-AS1 group compared with the si-NC group,and lower in the pc3.1-MSC-AS1 group compared with the vector group after overexpression of MSC-AS1.After silencing MSC-AS1 while inhibiting miR-520d-3p,the expression of miR-520d-3p and apoptosis rate of SiHa cells were in an order of si-NC+anti-miR-NC group<si-MSC-AS1+anti-miR-520d-3p group<si-MSC-AS1+anti-miR-NC group(P<0.05).MSC-AS1 targets miR-520d-3p,while miR-520d-3p targets ATAD2,and the apoptosis rate of SiHa cell in miR-520d-3p group were higher than in the miR-NC group(P<0.05).The ATAD2 protein levels,migration number,invasion number of SiHa cells were lower in the si-MSC-AS1 group than in the si-NC group,higher in the pc3.1-MSC-AS1group than in the vector group,ad lower in the miR-520d-3p group than in the miR-NC group,in an order of si-NC+anti-miR-NC group>si-MSC-AS1+anti-miR-520d-3p group>si-MSC-AS1+anti-miR-NC group(P<0.05).Conclusion Silencing MSC-AS1 may target ATAD2 through miR-520d-3p to inhibit the growth and metastasis of cervical cancer cells,and promote apoptosis.