Zhang Yonggang, Yang Hongling, Zhang Yipeng, Shi Junzhu, Chen Ronggui
Department of Clinical Laboratory, Shenzhen Longhua District Central Hospital, Guangdong Medical University, Shenzhen, 518110, Guangdong, China.
Department of Clinical Laboratory, Guangzhou Women and Children's Medical Centre, Guangzhou Medical University, 9, Jinsui Road, Guangzhou, 510623, China.
BMC Pregnancy Childbirth. 2020 Oct 27;20(1):652. doi: 10.1186/s12884-020-03345-5.
Preeclampsia is a severe disease in pregnant women, which is primarily managed by early screening and prevention. Circular RNAs (circRNAs) have increasingly been shown to be important biological regulators involved in numerous diseases. Further, increasing evidence has demonstrated that circRNAs can be used as diagnostic biomarkers. This study was conducted to evaluate the potential of circCRAMP1L, previously identified to be downregulated in preeclampsia, as a novel biomarker for predicting the development of preeclampsia.
We measured the expression of circCRAMP1L, which is reportedly relevant to trophoblast physiology, in plasma samples from 64 patients with preeclampsia and 64 age-, gestational age-, and body mass index-matched healthy pregnant women by qRT-PCR. MTT proliferation and transwell invasion assays revealed the biological role of circCRAMP1L in preeclampsia pathogenesis. RNA immunoprecipitation and dual-luciferase reporter assays clarified the mechanism underlying the biological function of circCRAMP1L in TEV-1 cells.
circCRAMP1L circulating levels were significantly lower in patients with preeclampsia (2.66 ± 0.82, △Ct value) than in healthy pregnant women (3.95 ± 0.67, △Ct value, p < 0.001). The area under the receiver operating characteristic curve for circCRAMP1L was 0.813. Univariate and multivariate analyses identified circCRAMP1L as an independent predictor of preeclampsia. Furthermore, when circCRAMP1L was utilised in combination with its target protein macrophage stimulating protein (MSP), the predictive performance increased, with an area under the receiver operating characteristic curve of 0.928 (95% CI 0.882-0.974), 80.0% sensitivity, and 80.0% specificity. The in vitro results indicated that circCRAMP1L regulates cell proliferation, and invasion via MSP and RON proteins. We investigated the molecular mechanisms of these effects. In vitro, relative to the control group, circCRAMP1L overexpression significantly enhanced cell proliferation; furthermore, trophoblast cell invasion increased proportionally with circCRAMP1L expression. RNA immunoprecipitation and luciferase reporter gene illustrated that circCRAMP1L participated in regulation of trophoblast cell by regulating MSP.
Reduced plasma levels of circCRAMP1L may be associated with an increased risk of preeclampsia, and circCRAMP1L may be a novel biomarker of preeclampsia risk.
子痫前期是孕妇的一种严重疾病，主要通过早期筛查和预防进行管理。环状RNA（circRNAs）越来越多地被证明是参与多种疾病的重要生物调节因子。此外，越来越多的证据表明circRNAs可作为诊断生物标志物。本研究旨在评估先前在子痫前期中被鉴定为下调的circCRAMP1L作为预测子痫前期发生的新型生物标志物的潜力。
我们通过qRT-PCR测量了64例子痫前期患者和64例年龄、孕周和体重指数匹配的健康孕妇血浆样本中circCRAMP1L的表达，据报道circCRAMP1L与滋养细胞生理学相关。MTT增殖和transwell侵袭试验揭示了circCRAMP1L在子痫前期发病机制中的生物学作用。RNA免疫沉淀和双荧光素酶报告试验阐明了circCRAMP1L在TEV-1细胞中生物学功能的潜在机制。
子痫前期患者的circCRAMP1L循环水平（2.66±0.82，△Ct值）显著低于健康孕妇（3.95±0.67，△Ct值，p<0.001）。circCRAMP1L的受试者工作特征曲线下面积为0.813。单因素和多因素分析确定circCRAMP1L是子痫前期的独立预测因子。此外，当circCRAMP1L与其靶蛋白巨噬细胞刺激蛋白（MSP）联合使用时，预测性能提高，受试者工作特征曲线下面积为0.928（95%CI 0.882-0.974），敏感性为80.0%，特异性为80.0%。体外结果表明，circCRAMP1L通过MSP和RON蛋白调节细胞增殖和侵袭。我们研究了这些作用的分子机制。在体外，相对于对照组，circCRAMP1L过表达显著增强细胞增殖；此外，滋养层细胞侵袭随circCRAMP1L表达成比例增加。RNA免疫沉淀和荧光素酶报告基因表明，circCRAMP1L通过调节MSP参与滋养层细胞的调控。
血浆中circCRAMP1L水平降低可能与子痫前期风险增加有关，circCRAMP1L可能是子痫前期风险的新型生物标志物。
