环状RNAhsa_circ_0005579在子痫前期患者胎盘中的表达及对滋养层细胞增殖、迁移及侵袭的影响
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E-mail:jiaruizhe2016@163.com

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R714.245

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国家自然科学基金(81971393)


The expression of circular RNA hsa_circ_0005579 in the placenta of patients with preeclampsia and its effect on the proliferation,migration and invasion of trophoblast cells
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    摘要:

    目的:探究环状RNA hsa_circ_0005579作为子痫前期发展的新生物标志物的潜力,观察其在子痫前期患者胎盘中的表达及其对滋养层细胞增殖、迁移及侵袭的影响。方法:通过实时定量聚合酶链式反应(qRT⁃PCR)检测20例子痫前期患者和 20例健康妊娠对照者胎盘组织中hsa_circ_0005579的表达,收集参与者的临床信息。采用核糖核酸酶R(RNase R)消化分析、 琼脂糖凝胶电泳及Sanger测序对hsa_circ_0005579进行鉴定,通过敲除siRNA转染HTR⁃8/SVneo细胞,利用CCK⁃8法、克隆形成分析、EdU实验检测hsa_circ_0005579对滋养层细胞增殖的影响,采用Transwell分析、划痕实验检测hsa_circ_0005579对滋养层细胞迁移及侵袭的影响。同时,蛋白质印迹(Western blot)分析用于检测迁移标志物的蛋白质水平。结果:hsa_circ_ 0005579在子痫前期患者胎盘中有显著的高表达。此外,与si⁃NC组相比,干扰hsa_circ_0005579的表达细胞活力明显增强、克隆形成数目增多、细胞增殖率增加、迁移及侵袭数目显著增多、细胞迁移率提高以及迁移标志物基质金属蛋白酶2(MMP2)、基质金属蛋白酶 9(MMP9)蛋白水平提升。结论:环状 RNA hsa_circ_0005579 在子痫前期患者胎盘中高表达,沉默 hsa_circ_ 0005579后HTR⁃8/SVneo的增殖、迁移及侵袭能力增强,有望为子痫前期的早期诊断和干预提供新的靶标。

    Abstract:

    Objective:To explore the potential of the circular RNA hsa_circ_0005579 as a new biomarker for the development of pre ⁃eclampsia,and to observe its expression in the placenta of patients with prE-eclampsia and its effect on the proliferation,migration and invasion of trophoblast cells. Methods:The expression of hsa_circ_0005579 was detected in placental tissues obtained from 20 women with pre ⁃ eclampsia and 20 healthy pregnancy controls using real ⁃time quantitative polymerase chain reaction(qRT ⁃ PCR). Clinical information of participants was collected. Ribonuclease R(RNase R)digestion analysis,agarose gel electrophoresis and Sanger sequencing were used to verify hsa_circ_0005579. HTR⁃8/SVneo cells were transfected with hsa_circ_0005179 siRNA,cell counting kit⁃8(CCK⁃8)assay,colony formation assay,EdU test were performed to detect the effect of hsa_circ_0005579 on the proliferation of trophoblast cells. Transwell assay and wound healing assay were used to detect the effect of hsa_circ_0005579 on the migration and invasion of trophoblast cells. Meanwhile,Western blot analysis was used to detect the protein level of migration markers. Results: Hsa_circ_0005579 had a significant high expression in the placenta of PE patients. In addition,compared with the si ⁃NC group,the cell viability was significantly increased,the number of clone formation grew,the cell proliferation rate increased,and the number of migration and invasion did rise,the cell migration rate was upregulated,and the protein levels of migration markers matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9)was enhanced in the si⁃circ_0005579 group. Conclusion:CircularRNA hsa_circ_0005579 is highly expressed in the placenta of patients with pre ⁃ eclampsia,and the proliferation,migration and invasion ability of HTR ⁃ 8/SVneo is enhanced after silencing hsa_circ_0005579. It is expected to provide new targets for early diagnosis and intervention of prE-eclampsia.

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高明明,俞兆儿,姚丹,仲红,贾瑞喆.环状RNAhsa_circ_0005579在子痫前期患者胎盘中的表达及对滋养层细胞增殖、迁移及侵袭的影响[J].南京医科大学学报(自然科学版),2022,42(8):1112-1118

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  • 在线发布日期: 2022-08-07
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