lncRNA HOXA-AS2靶向miR-520a-3p调控卵巢癌细胞增殖、迁移和侵袭

lncRNA HOXA-AS2 regulates proliferation, migration and invasion of ovarian cancer cells via targeting miR-520a-3p

目的:探究长链非编码RNA HOXA-AS2(lncRNA HOXA-AS2)与微小RNA-520a-3p(miR-520a-3p)之间的靶向关系及其对卵巢癌SKOV3细胞增殖、迁移和侵袭的影响。方法:q PCR检测lncRNA HOXA-AS2与miR-520a-3p在多种卵巢癌细胞(SKOV3、HO8910、OVCAR3细胞)及正常卵巢上皮细胞HOSE中的表达水平。生物信息学手段预测HOXA-AS2与miR-520a-3p之间的靶向关系并用双荧光素酶报告基因实验验证。将si-HOXA-AS2、miR-520a-3p mimic、anti-miR-520a-3p和相应对照片段分别或共转染SKOV3细胞,MTT、Transwell和Western blotting法分别检测各组SKOV3细胞增殖、迁移、侵袭及相关蛋白(CyclinD1、p21、p27、MMP-2、MMP-9、MMP-14)表达情况。结果:与HOSE细胞相比,多种卵巢癌细胞中HOXA-AS2均呈高表达(均P<0.05)、miR-520a-3p均呈低表达(均P<0.05);HOXA-AS2可靶向下调miR-520a-3p的表达。si-HOXA-AS2和miR-520a-3p mimics组SKOV3细胞的增殖、迁移及侵袭能力均较对照组均显著降低(均P<0.01),且p21、p27蛋白表达显著升高,而CyclinD1、MMP-2、MMP-9、MMP-14蛋白表达显著减少(均P<0.01);si-HOXA-AS2+anti-miR-520a-3p组SKOV3细胞增殖、迁移及侵袭能力较si-HOXA-AS2和si-HOXA-AS2+anti-miR-NC组显著增强(均P<0.05)。结论:lncRNA HOXA-AS2通过靶向抑制miR-520a-3p表达进而增强卵巢癌SKOV3细胞的增殖、迁移及侵袭能力。

Objective: To explore the targeting relationship between long-chain noncoding RNA HOXA-AS2(lncRNA HOXA-AS2 and microRNA-520a-3p(miR-520a-3p)) and their effects on the proliferation, migration and invasion of ovarian cancer SKOV3 cells.Methods:qPCR was used to detect the expression levels of lncRNA HOXA-AS2 and miR-520a-3p in various ovarian cancer cell lines(SKOV3, HO8910, OVCAR3 cells) and normal ovarian epithelial cell line HOSE. Bioinformatics methods were used to predict the targeting relationship between HOXA-AS2 and miR-520a-3p, which was then verified by Dual luciferase reporter gene assay.si-HOXA-AS2, miR-520a-3p mimic, anti-miR-520a-3p and corresponding control fragments were transfected into SKOV3 cells separately or in combination. MTT, Transwell and Western blotting were used to detect the proliferation, migration, invasion and expressions of related proteins(CyclinD1, p21, p27, MMP-2, MMP-9, MMP-14) of SKOV3 cells in each group. Results: Compared with HOSE cells, HOXA-AS2 was over-expressed while miR-520a-3p was under-expressed in ovarian cancer cell lines(all P<0.05).HOXA-AS2 could targetedly down-regulate the expression of miR-520a-3p. Compared with the NC group, the proliferation, migration and invasion of SKOV3 cells in the si-HOXA-AS2 and miR-520a-3p mimics groups were significantly reduced(all P<0.01),and the protein expressions of p21 and p27 were significantly increased, while protein expressions of CyclinD1, MMP-2, MMP-9,MMP-14 were significantly reduced(all P<0.01). The proliferation, migration and invasion of SKOV3 cells in the si-HOXA-AS2+antimiR-520a-3p group were significantly enhanced compared with those in si-HOXA-AS2 and si-HOXA-AS2+anti-miR-NC groups(all P<0.05). Conclusion: lncRNA HOXA-AS2 enhances the proliferation, migration and invasion of ovarian cancer SKOV3 cells by targetedly inhibiting the expression of miR-520a-3p.