lncRNA CASC11靶向miR-498对滋养层细胞迁移侵袭的影响

Effects of lncRNA CASC11 targeting miR-498 on the migration and invasion of trophoblast cells

目的 探究长链非编码RNA(lncRNA)癌症易感性候选物11(CASC11)靶向miR-498对滋养层细胞迁移侵袭的影响。方法 收集本院22份PE患者胎盘组织,28份正常妊娠女性胎盘组织,实时荧光定量PCR(qRT-PCR)检测lncRNACASC11及miR-498表达;将HTR8/Svneo细胞分成对照组、pcDNA组、pcDNA-CASC11组、pcDNACASC11+miR-NC组、pcDNA-CASC11+miR-498 mimic组;3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法、划痕实验及Transwell实验分别测定HTR8/Svneo细胞增殖、迁移及侵袭能力,Westernblot检测细胞基质金属蛋白酶(MMP)2、c-Myc、MMP-9表达;双荧光素酶报告基因检测lncRNACASC11及miR-498靶向关系。结果 与正常胎盘组织相比,PE胎盘组织中lncRNACASC11表达显著降低,miR-498表达显著增加(P<0.01)。与对照组相比,pcDNA-CASC11组lncRNA CASC11表达、细胞存活率、迁移率、侵袭细胞数、c-Myc、MMP-2及MMP-9表达显著增加,miR-498表达显著降低(P<0.01);与pcDNA-CASC11组相比,pcDNA-CASC11+miR-498 mimic组细胞存活率、迁移率、侵袭细胞数、c-Myc、MMP-2及MMP-9表达显著降低,miR-498表达显著增加(P<0.01)。lncRNACASC11与miR-498存在靶向关系。结论 lncRNA CASC11通过靶向抑制miR-498表达促进HTR8/Svneo细胞迁移侵袭。

Objective To explore the effects of long non-coding RNA(lncRNA) cancer susceptibility candidate 11(CASC11) targeting microRNA-498(miR-498) on the migration and invasion of trophoblast cells. Methods 22 placental tissues of PE patients and 28 placental tissues of normal pregnant women in our hospital were collected. Quantitative real-time PCR(qRT-PCR) was used to detect the expression of lncRNA CASC11 and miR-498. HTR8/Svneo cells were divided into control group, pcDNA group, pcDNA-CASC11 group, pcDNA-CASC11+miR-NC group, and pcDNA-CASC11+miR-498mimic group. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) method, scratch test and Transwell test were used to measure the migration and invasion abilities of HTR8/Svneo cells respectively. Western blot was used to detect the expression of matrix metalloproteinase(MMP) 2 and c-Myc and MMP-9 in cells. Dual luciferase reporter gene was used to detect the targeting relationship between lncRNA CASC11 and miR-498. Results Compared with normal placental tissue, the expression of lncRNA CASC11 in PE placental tissue significantly reduced, and the expression of miR-498 significantly increased(P<0.01). Compared with the control group, the expression of lncRNA CASC11, cell survival rate, migration rate,number of invasive cells, expression of c-Myc, MMP-2 and MMP-9 in the pcDNA-CASC11 group significantly increased, and the expression of miR-498 significantly reduced(P<0.01). Compared with the pcDNA-CASC11 group, the cell survival rate,migration rate, number of invasive cells, expression of c-Myc, MMP-2 and MMP-9 in the pcDNA-CASC11 group significantly decreased, and the expression of miR-498 significantly increased(P<0.01). There was a targeting relationship between lncRNA CASC11 and miR-498. Conclusion LncRNA CASC11 promotes the migration and invasion of HTR8/Svneo cells through targeting the inhibition of miR-498 expression.