lncRNA PSMA3-AS1靶向miR-3619-5p调控宫颈癌SiHa细胞增殖、迁移及侵袭的分子机制研究

lncRNA PSMA3-AS1 Regulate the Proliferation,Migration and Invasion of Cervical Cancer Cells by Targeting miR-3619-5p

目的探讨lncRNA PSMA3-AS1调控宫颈癌细胞增殖、迁移及侵袭的分子机制.方法收集2020年3月至2021年6月西安医学院第二附属医院收治的42例宫颈癌患者的癌组织及其癌旁组织标本,采用qRT-PCR法检测宫颈癌组织、癌旁组织、人宫颈上皮永生化细胞H8、与人宫颈癌细胞系SiHa、HeLa、Cas-ki中lncRNA PSMA3-AS1、miR-3619-5p的表达量;以SiHa细胞为研究对象,分组为:si-NC组、si-lncRNA PSMA3-AS1组、miR-NC组、miR-3619-5p组、anti-miR-NC+si-lncRNA PSMA3-AS1组、anti-miR-3619-5p+si-lncRNA PSMA3-AS1组;MTT法与Transwells实验分别检测每组SiHa细胞的增殖活力、迁移及侵袭能力;双荧光素酶报告实验检测miR-3619-5p过表达对野生型载体lncRNA PSMA3-AS1-WT、突变型载体lncRNA PS-MA3-AS1-MUT荧光素酶活性的影响;Western印迹检测MMP2、MMP9蛋白表达量.结果宫颈癌组织中lncRNA PSMA3-AS1的表达量比癌旁组织增加(P<0.01),miR-3619-5p的表达量比癌旁组织减少(P<0.01);与H8细胞比较,SiHa、HeLa、Caski细胞中lncRNA PSMA3-AS1的表达量升高(P<0.01),miR-3619-5p的表达量降低(P<0.01);与si-NC组比较,si-lncRNA PSMA3-AS1组细胞活力和MMP2、MMP9蛋白水平降低(P<0.05),迁移及侵袭细胞数减少(P<0.05);与miR-NC组比较,miR-3619-5p组细胞活力和MMP2、MMP9蛋白水平降低(P<0.01),迁移及侵袭细胞数减少(P<0.05);miR-3619-5p过表达可抑制lncRNA PSMA3-AS1-WT的荧光素酶活性(P<0.01),而未能影响lncRNA PSMA3-AS1-MUT的荧光素酶活性;与anti-miR-NC+si-lncRNA PSMA3-AS1组比较,anti-miR-3619-5p+si-lncRNA PSMA3-AS1组细胞活力和MMP2、MMP9蛋白水平升高(P<0.01),迁移及侵袭细胞数增多(P<0.01).结论干扰lncRNA PSMA3-AS1表达可通过促进miR-3619-5p而降低宫颈癌细胞增殖、迁移及侵袭能力.

ObjectiveTo explore the molecular mechanism of lncRNA PSMA3-AS1 on regulating the proliferation,migration and invasion of cervical cancer cells.MethodsCollect cancer tissue and paracancerous tissue specimens of 42 patients with cervical cancer admitted to the Second Affiliated Hospital of Xi'an Medical University from March 2020 to June 2021.The qRT-PCR method was used to detect the expression levels of lncRNA PSMA3-AS1,miR-3619-5p in cervical cancer tissues,adjacent tissues,human cervical epithelial immortalized cells H8,and human cervical cancer cell lines SiHa,HeLa,and Cashi.Taking SiHa cells as the research object,they were randomly grouped into 6 groups:si-NCgroup,si-lncRNA PSMA3-AS1 group,miR-NC group,miR-3619-5p group,anti-miR-NC+si-lncRNA PSMA3-AS1 group,anti-miR-3619-5p+si-lncRNA PSMA3-AS1 group.The proliferation,migration and invasion abilities of SiHa cells in each group were assayed by MTT assay and transwell assay respectively.The dual luciferase reporter assay was used to determine the effect of miR-3619-5p overexpression on the luciferase activity of wild-type vector lncRNA PSMA3-AS1-WT and mutant vector lncRNA PSMA3-AS1-MUT.Western blotting was used to detect the protein expression levels of MMP2 and MMP9.ResultsThe expression level of IncRNA PSMA3-AS1 in cervical cancer tissues was higher than that in the adjacent tissues(P<0.01),and the expression level of miR-3619-5p was lower than that in the adjacent tissues(P<0.01).Compared with the H8 cells,the expression level of IncRNA PSMA3-AS1 in SiHa,HeLa,and Caski cells was increased(P<0.01),while the expression level of miR-3619-5p was decreased(P<O.01).Compared with the si-NC group,the cell viability,and the protein expression levels of MMP2,MMP9 in the si-lncRNA PSMA3-AS1 group were decreased(P<0.05),and the numbers of migrated and invaded cells were decreased(P<0.05).Compared with the miR-NC group,the cell viability and the protein expression levels of MMP2,MMP9 in the miR-3619-5p group were decreased(P<0.01),and the numbers of migrated and invaded cells were decreased(P<0.01).Overexpression of miR-3619-5p could inhibit the luciferase activity of lncRNA PSMA3-AS1-WT(P<0.01),but failed to affect the luciferase activity of IncRNA PSMA3-AS1-MUT.Compared with the anti-miR-NC+si-lncRNA PSMA3-AS1 group,the cell viability,and the protein expression levels of MMP2,MMP9 in the anti-miR-3619-5p+si-lncRNA PSMA3-AS1 group were increased(P<0.01),and the numbers of migrated and invaded cells were increased(P<0.01).Conclusion Interfering with the expression of lncRNA PSMA3-AS1 could reduce the proliferation,migration and invasion of cervical cancer cells by targeting miR-3619-5p.