lncRNAs ABHD11-AS1靶向miR-133a上调EGFR表达促进卵巢癌细胞增殖

LncRNAs ABHD11-AS1 Targets miR-133a to Upregulate EGFR Expr-ession and Promote Ovarian Cancer Cell Proliferation

目的研究lncRNAs ABHD11-AS1在卵巢癌细胞和输卵管上皮细胞中的表达,探究其对卵巢癌细胞增殖的影响和作用机制。方法Real-time PCR检测细胞中lncRNAs ABHD11-AS1和miR-133a的表达,双荧光素酶实验明确二者之间的调控作用;用NC siRNA、ABHD11-AS1 siRNA以及NC mimic、miR-133a mimic和NC inhibitor、miR-133a inhibitor分别转染细胞,CCK-8检测细胞增殖;Real-time PCR以及Western Blot实验检测细胞中EGFR的表达。结果结果表明,lncRNAs ABHD11-AS1在TOV-112D和CaVO3卵巢癌细胞系中的表达水平明显高于TEC细胞,且下调lncRNAs ABHD11-AS1可明显抑制卵巢癌细胞的增殖;ABHD11-AS1与miR-133a的表达呈负相关。双荧光素酶结果显示,转染miR-133a mimic和野生型ABHD11-AS1片段的细胞中荧光素酶的活性显著降低;与输卵管上皮细胞相比,miR-133a在卵巢癌细胞系的表达水平更低。转染miR-133a mimic会显著上调上述两种细胞中miR-133a的表达同时还抑制其增殖和EGFR表达。此外,下调miR-133a可明显促进卵巢癌细胞的增殖以及增加EGFR表达。与此相反,下调lncRNAs ABHD11-AS1明显降低EGFR表达。结论lncRNAs ABHD11-AS1通过调控miR-133a,上调其下游靶基因EGFR的表达从而促进卵巢癌的发生发展。

Objective To investigate the expression of LncRNAs ABHD11-AS1 in ovarian cancer cell and oviductal epithelial cell and explore the effect of LncRNAs ABHD11-AS1 on ovarian cancer cell proliferation.Methods The level of LncRNAs ABHD11-AS1 and miR-133a was detected by Real-time PCR,and interaction between them was confirmed by Dual-Luciferase reporter assay(DR).Cells were transfected with NC siRNA,ABHD11-AS1 as well as siRNA NC mimic,miR-133a mimic and NC inhibitor and miR-133a inhibitor,respectively,and the proliferation was detected by Cell Counting Kit-8(CCK-8)assay.Real-time PCR and western blotting were performed to detect the EGFR expression.Results Downregulation of lncRNAs ABHD11-AS1 could significantly inhibit the proliferation of ovarian cancer cells.Moreover,lncRNAs ABHD11-AS1 was negatively correlated with the expression of miR-133a.DR results showed that luciferase activity was significantly reduced in cells transfected with miR-133a mimic and wild-type ABHD11-AS1 fragments;and the level of miR-133a was lower in ovarian cancer cell compared to oviductal epithelial cells.Transfection of miR-133a mimic significantly upregulated miR-133a expression in both of these cells and inhibited their proliferation and EGFR expression.In addition,downregulating miR-133a significantly promoted the proliferation of ovarian cancer cells and increased the EGFR expression,while downregulating LncRNAs ABHD11-AS1 significantly decreased the EGFR expression.Conclusion LncRNAs ABHD11-AS1 promoted the malignant behaviors of ovarian cancer by targeting miR-133a to upregulate its downstream target gene EGFR.