早期生长反应因子1特异诱骗寡核苷酸下调富含半胱氨酸蛋白61抑制大鼠血管平滑肌细胞迁移
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国家自然科学基金资助(30871074)


Decoy Oligodecoxynucleotides Targeting Early Growth Response Factor-1 Downregulating Cysteine-rich 61 to Inhibit Rat Vascular Smooth Muscle Cells Migration
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    目的观察早期生长反应因子1特异诱骗寡核苷酸对体外培养的大鼠血管平滑肌细胞中早期生长反应因子1和富含半胱氨酸蛋白61表达的影响,初步探讨早期生长反应因子1特异诱骗寡核苷酸抑制血管平滑肌细胞迁移的机制。方法体外培养血管平滑肌细胞并转染早期生长反应因子1特异诱骗寡核苷酸,采用逆转录聚合酶链反应和免疫组织化学法检测转染后不同时间点对照组与实验组血管平滑肌细胞中早期生长反应因子1及富含半胱氨酸蛋白61的mRNA和蛋白的表达变化,划痕法测定血管平滑肌细胞迁移距离。结果早期生长反应因子1及富含半胱氨酸蛋白61的mRNA和蛋白在对照组、诱骗组及诱骗对照组中均有表达。在不同时间点,诱骗组早期生长反应因子1及富含半胱氨酸蛋白61的mRNA和蛋白的表达均低于对照组(p<0.05)。转染后48 h,对照组、诱骗组及诱骗对照组血管平滑肌细胞的迁移距离分别为105.23±7.81 μm、58.65±12.68 μm、106.47±7.60 μm,诱骗组血管平滑肌细胞的迁移距离明显低于其他两组(p<0.01)。结论对体外培养的血管平滑肌细胞转染早期生长反应因子1特异诱骗寡核苷酸可以抑制早期生长反应因子1及富含半胱氨酸蛋白61的mRNA和蛋白表达,从而抑制血管平滑肌细胞的迁移。

    Abstract:

    AimTo observe the influence of the early growth response factor-1 (Egr-1) decoy oligodeoxynucleotide (decoy ODNs) on the expression of Egr-1 and cysteine-rich protein 61 (CYR61/CCN1) in rat vascular smooth muscle cells (VSMCs) cultured in vitro and investigate the mechanism about Egr-1 decoy ODNs inhibiting VSMCs migration.MethodsCultured the VSMCs in vitro and transfected Egr-1 decoy ODNs.Detected the mRNA level and the protein expression of Egr-1 and CCN1 using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical method at the different time points of normal control group (normal cultured cells) and the decoy group (transfected Egr-1 decoy ODN).The migration lenth of each group was observed by wound-healing assay.ResultsThe results of RT-PCR and cell immunohistochemistry analysis showed that there was certain expression of Egr-1 and CCN1 mRNA and protein in these three groups.At the different time points, the level of Egr-1 and CCN1 mRNA and protein were decreased obviously in the decoy group compared with the normal control group (p<0.05).Wound-healing assay showed that the migration length of the control group, decoy and decoy SCR was 105.23±7.81 μm, 58.65±12.68 μm and 106.47±7.60 μm respectively.The migration length of the decoy group is much shorter than the other groups (p<0.01).ConclusionTransfecting the decoy ODNs of Egr-1 to the arterial VSMCs of rat in vitro can decrease the expression of Egr-1 and CCN1, to achieve the purpose of inhibiting cell migration.

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徐否,刘闺男,韩伟.早期生长反应因子1特异诱骗寡核苷酸下调富含半胱氨酸蛋白61抑制大鼠血管平滑肌细胞迁移[J].中国动脉硬化杂志,2012,20(6):487~493.

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  • 收稿日期:2011-06-24
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