LncRNA PTPRG-AS1通过靶向miR-5590-3p调控乳腺癌细胞增殖、迁移和侵袭的研究

LncRNA PTPRG-AS1 regulates breast cancer cell proliferation,migration and invasion by targeting miR-5590-3p

目的探讨LncRNA PTPRG-AS1对乳腺癌细胞增殖、迁移和侵袭的影响及可能机制。方法以正常乳腺上皮细胞MCF-10A为对照,qRT-PCR法检测乳腺癌细胞系(MCF-7、T47D和BT549)中 LncRNA PTPRG-AS1 和 miR-5590-3p 表达。分别转染 si-LncRNA PTPRG-AS1、miR-5590-3p mimics、共转染si-LncRNA PTPRG-AS1与anti-miR-5590-3p至乳腺癌MCF-7细胞,用CCK-8法检测细胞增殖,Transwell检测检测细胞迁移和侵袭,蛋白质印迹法检测细胞中CyclinD1、MMP2和MMP9蛋白表达。双荧光素酶报告基因实验验证LncRNA PTPRG-AS1和miR-5590-3p的调控关系。结果与MCF-10A细胞比较,乳腺癌细胞系(MCF-7、T47D和BT549)中LncRNA PTPRG-AS1表达升高,差异有统计学意义(P<0.05),miR-5590-3p表达降低,差异有统计学意义(P<0.05)。敲减LncRNA PTPRG-AS1或过表达miR-5590-3p后,MCF-7细胞增殖活性、迁移数、侵袭数及细胞中CyclinD1、MMP2和MMP9蛋白表达均降低,差异有统计学意义(P<0.05)。LncRNA PTPRG-AS1靶向负调控miR-5590-3p。敲减miR-5590-3p逆转了敲减LncRNA PTPRG-AS1对MCF-7细胞增殖、迁移和侵袭的影响。结论 LncRNA PTPRG-AS1可能靶向下调miR-5590-3p促进乳腺癌细胞增殖、迁移和侵袭。

Objective To explore the effect of LncRNA PTPRG-AS1 on the proliferation,migration and invasion of breast cancer cells and its possible mechanism.Methods With normal breast epithelial cells MCF-10 A as a control,the expression of LncRNA PTPRG-AS 1 and miR-5590-3 p in breast cancer cell lines(MCF-7,T47 D and BT549) was detected by qRT-PCR.Breast cancer MCF-7 cells were transfected with si-LncRNA PTPRG-AS 1 or miR-5590-3 p mimics,or co-transfected with si-LncRNA PTPRGAS1 and anti-miR-5590-3 p,and CCK-8 method was used to detect cell proliferation;Trans well was used to detect cell migration and invasion.The protein expression of CyclinDl,MMP2 and MMP9 in cells were detected by Western blotting.The dual luciferase reporter gene experiment verified the regulatory relationship between LncRNA PTPRG-AS1 and miR-5590-3 p.Results Compared with MCF-10 A cells,the expression of LncRNA PTPRG-AS1 in breast cancer cell lines(MCF-7,T47 D and BT549) was increased(P<0.05),but the expression of miR-5590-3 p decreased(P<0.05).After knocking down LncRNA PTPRG-AS1 or overexpressing miR-5590-3 p,the proliferation activity of MCF-7 cells,the number of migration and invasion,and the protein expression of CyclinDl,MMP2 and MMP9 were all reduced(P<0.05).LncRNA PTPRG-AS1 could target and negatively regulate miR-5590-3 p.Knocking down miR-5590-3 p reversed the effect of knocking down LncRNA PTPRG-AS1 on the proliferation,migration and invasion of MCF-7 cells.Conclusion LncRNA PTPRG-AS1 may target down-regulation of miR-5590-3 p to promote breast cancer cell proliferation,migration and invasion.