中肾因子Midkine对卵巢癌中非编码RNA表达的影响

Effect of midkine on expression of non-coding RNA in ovarian cancer

目的探讨肿瘤相关抗原中肾因子(midkine,MK)调控的长链非编码RNA(lncRNAs)和微小RNA(miRNAs)对卵巢癌的临床应用价值。方法采用基因芯片(miRNAs芯片和lncRNAs芯片)分析MK过表达的SKOV3-MK细胞及其SKOV3-Con对照细胞中差异的miRNAs和lncRNAs,筛选卵巢癌潜在的生物学标志物,并在卵巢癌患者血清和卵巢癌组织中用RT-qPCR进一步验证,结合临床数据,分析其临床应用价值。结果与对照SKOV3-Con细胞相比,MK过表达可以显著促进SKOV3细胞中11个miRNAs和7个lncRNAs的表达(比值>3倍,P<0.01),下调8个miRNAs和13个lncRNAs的表达(比值<0.3,P<0.01)。RT-qPCR验证结果表明,相对于对侧良性卵巢组织,卵巢癌组织中miR489的表达水平显著降低,而HOTAIR则显著升高(P<0.05);卵巢癌患者血清中HOTAIR的表达水平显著高于同年龄段健康人对照组(0.036±0.024 vs 0.019±0.020,P=0.002)。此外,当cut-off值为0.0176时,HOTAIR作为卵巢癌血清筛查指标的AUC ROC为0.749,特异性为66.7%,敏感性为75.6%。结论长链非编码RNA HOTAIR可以作为卵巢癌血清的筛查标志物之一。

Objective To explore the correlation of non-coding RNA and the tumor-associated antigen midkine(MK)in SKOV3cells and the clinical significance for diagnosis of ovarian cancer.Methods The Agilent′s gene chips(miRNAs chip and lncRNAs chip)were used to analyze the differential expression of miRNAs and lncRNAs in both MK-overexpressing SKOV3-MK cells and the control SKOV3-Con cells to screen the potential biomarkers in ovarian cancer.The clinical significance of midkine in the serum and tissues samples was analyzed for the patients with ovarian cancer by quantitative PCR combined with clinical data.Results Compared with control SKOV3-con cells,MK overexpression significantly promoted the expressions of 11 miRNAs and 7 lncRNAs in SKOV3 cells(P<0.01,ratio>3 fold),reduced the expressions of 8 miRNAs and 13 lncRNAs(P<0.01,ratio<0.3).Results of qPCR showed that the expression level of miR489 was significantly lower in ovarian cancer tissues than that of the contralateral normal ovarian tissues,while HOTAIR was significantly elevated(P<0.05).The expression level of HOTAIR in the serum of ovarian cancer patients was significantly higher than that in healthy controls group with same age(0.036±0.024 vs 0.019±0.020,P=0.002).ROC curve analysis of HOTAIR showed that the specificity was 66.7%,the sensitivity was 75.6%and the AUC value was 0.749 as a marker for serum detection of ovarian cancer when the cutoff value was 0.0176.Conclusion Long-chain non-coding RNA HOTAIR may be served as a potential biomarker in serum of ovarian cancer patients.