lncRNA VPS9D1-AS1调节miR-331-3p/SERP1轴对宫颈癌细胞增殖、凋亡和侵袭的影响
Effects of lncRNA VPS9D1-AS1 on proliferation,apoptosis,and invasion of cervical cancer cells by regulating the miR-331-3p/SERP1 axis
师媛 1张赞 2陈璐 1周满红1
作者信息
- 1. 西北妇女儿童医院,陕西西安 716000
- 2. 延安大学咸阳医院,陕西咸阳 712000
- 折叠
摘要
目的 探讨lncRNA VPS9D1-AS1对宫颈癌细胞恶性生物学行为的影响及作用机制.方法 分析宫颈癌细胞和宫颈癌组织中lncRNA VPS9D1-AS1表达情况;将SiHa细胞分为si-NC组、si-VPS9D1-AS1组、si-VPS9D1-AS1+inhibitor-NC 组、si-VPS9D1-AS1+miR-331-3p inhibitor 组和 control 组,qRT-PCR 检测转染效率;通过集落形成实验、流式细胞术、Transwell实验、宫颈癌细胞肿瘤异种移植生长等一系列功能实验研究lncRNA VPS9D1-AS1对宫颈癌的影响.采用荧光素酶报告基因法、蛋白质印迹法验证lncRNA VPS9D1-AS1与miR-331-3p、miR-331-3p与SERP1的靶向关系.结果 VPS9D1-AS1在宫颈癌组织和细胞中上调表达(P＜0.05);与si-NC组比较,si-VPS9D1-AS1组SiHa细胞集落形成数、PCNA、SERP1表达、迁移与侵袭细胞数、MMP-2、MMP-9表达降低,细胞凋亡率、Cleaved Caspase-3表达升高(P＜0.05);下调miR-331-3p可逆转VPS9D1-AS1敲低对SiHa细胞恶性行为的抑制作用(P＜0.05);双荧光素酶报告基因实验证实miR-331-3p与lncRNA VPS9D1-AS1、miR-331-3p与SERP1存在靶向调控关系(P＜0.05);裸鼠实验表明,体内抑制VPS9D1-AS1表达可显著降低移植瘤质量、体积、SERP1表达,升高miR-331-3p表达(P＜0.05).结论 lncRNA VPS9D1-AS1在宫颈癌组织和细胞中上调表达,敲低VPS9D1-AS1通过调节miR-331-3p/SERP1轴,抑制宫颈癌细胞的恶性进展.
Abstract
Objective To investigate the effect and mechanism of lncRNA VPS9D1-AS1 on the malignant biological behavior of cervical cancer cells.Methods The expression of lncRNA VPS9D1-AS1 in cervical cancer cells and cervical can-cer tissues was analyzed.SiHa cells were separated into si-NC group,si-VPS9D1-AS1 group,si-VPS9D1-AS1+inhibitor NC group,and si-VPS9D1-AS1+miR-331-3p inhibitor group and control group.qRT-PCR was applied to detect transfection effi-ciency.A series of functional experiments such as colony formation assay,flow cytometry,Transwell assay and cervical cancer cell xenograft growth were applied to study the effect of lncRNA VPS9D1-AS 1 on cervical cancer.Luciferase reporter gene method and Western blotting method were applied to verify the targeting relationship between lncRNA VPS9D1-AS1 and miR-331-3p,and between miR-331-3p and SERP1.Results The expression of VPS9D1-AS1 was up-regulated in cervical cancer tissues and cells(P＜0.05).Compared with the si-NC group,the number of SiHa cell colony formation,the expression of PCNA and SERP1,the number of migration and invasion cells,the expression of MMP-2 and MMP-9 in the si-VPS9D1-AS1 group were decreased,and the apoptosis rate and the expression of Cleaved Caspase-3 were increased(P＜0.05).Downregulation of miR-331-3p was able to reverse the inhibitory effect of VPS9D1-AS1 knockdown on the malig-nant behavior of SiHa cells(P＜0.05).Dual luciferase reporter gene assay confirmed that miR-331-3p and lncRNA VPS9D1-AS1,miR-331-3p and SERP1 had a targeted regulatory relationship(P＜0.05).Nude mouse experiments showed that inhibiting the expression of VPS9D1-AS1 in vivo was able to obviously reduce the mass,volume,and SERP1 expression of transplanted tumors,and increase the expression of miR-331-3p(P＜0.05).Conclusion In cervical cancer tissues and cells,lncRNA VPS9D1-AS1 is up-regulated.Knockdown of VPS9D1-AS1 blocks the malignant progression of cervical cancer cells through regulating the miR-331-3p/SERP1 axis.
关键词
lncRNA/VPS9D1-AS1/miR-331-3p/SERP1轴/宫颈癌/增殖/凋亡/侵袭Key words
lncRNA VPS9D1-AS1/MiR-331-3p/SERP1 axis/cervical cancer/proliferation/apoptosis/invasion引用本文复制引用
出版年
2024
NETL
NSTL
万方数据