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首页> 外文期刊>Biochemical Genetics >Retrovirus-Mediated Transfection of the Tissue-type Plasminogen Activator Gene Results in Increased Thrombolysis of Blood Clots
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Retrovirus-Mediated Transfection of the Tissue-type Plasminogen Activator Gene Results in Increased Thrombolysis of Blood Clots

机译：逆转录病毒介导的组织型纤溶酶原激活剂基因导致血栓溶栓增加

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摘要

Tissue-type plasminogen activator (tPA) is involved in the lysis of blood clots. In this study, we attempted to target thrombolysis and enhance blood clot lysis by generating a construct (pLEGFP-N1-tPA) to integrate tPA gene into the genome of different cell lines. pLEGFP-N1-tPA construct was generated and used to target the tPA gene in different cell lines. The thrombolytic effects mediated by the supernatant from transfected HeLa cells and Linx cells were assessed using plasma thrombus plates. Furthermore, enhanced green fluorescent protein (EGFP), which was fused to the tPA gene in the pLEGFP-N1-tPA construct, was analyzed under the fluorescent microscope to assess tPA localization. We also monitored tPA activity and expression in the transfected cell lines. As part of the study, we successfully generated the pLEGFP-N1-tPA construct. The sequence of this construct was verified and the construct was subsequently used to generate the PT67/pLEGFP-N1-tPA cell line. The pLEGFP-N1-tPA construct was also used to transfect HeLa cells and Linx cells. We observed that supernatants from transfected cells were capable of lysing thrombi. In addition, tPA activity and tPA concentration were elevated in the latter supernatants and tPA was rapidly and stably expressed in the transfected cell lines. These results reveal a potentially important thrombolytic role for tPA-targeted gene therapy following cardiac valve replacement.
机译：组织型纤溶酶原激活剂（TPA）参与血栓的裂解。在这项研究中，我们试图通过产生构建体（Plegfp-N1-TPA）来诱导溶栓和增强血凝块裂解，以将TPA基因与不同细胞系的基因组。生成PlegFP-N1-TPA构建体并用于靶向不同细胞系中的TPA基因。使用血浆血栓平板评估由上清液介导的上清液和LINX细胞介导的溶血性效应。此外，在荧光显微镜下分析了增强的绿色荧光蛋白（EGFP），其与Plegfp-N1-TPA构建体中的TPA基因融合，以评估TPA定位。我们还监测转染细胞系中的TPA活动和表达。作为研究的一部分，我们成功地生成了PleGFP-N1-TPA构造。验证该构建体的序列，随后使用构建体来产生PT67 / PLEGFP-N1-TPA细胞系。 PLEGFP-N1-TPA构建体还用于转染HeLa细胞和Linx细胞。我们观察到转染细胞的上清液能够裂解血栓。另外，在后一种上清液中升高TPA活性和TPA浓度，并且TPA在转染的细胞系中快速且稳定地表达。这些结果揭示了TPA靶向基因治疗后心瓣术后的潜在重要的血栓性作用。

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  • 来源
    《Biochemical Genetics》 |2019年第2期|共14页
  • 作者

    Wu Xianhua; Gong Yongsheng; Ding Xuebing; Cheng Gang; Yan Weiya; She Xiaowei; Wang Changxing; Li Xia;

  • 作者单位

    Nanjing Med Univ Dept Thorac Cardiovasc Surg Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008;

    Nanjing Med Univ Dept Thorac Cardiovasc Surg Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008;

    Nanjing Med Univ Dept Thorac Cardiovasc Surg Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008;

    Nanjing Med Univ Dept Thorac Cardiovasc Surg Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008;

    Nanjing Med Univ Dept Thorac Cardiovasc Surg Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008;

    Nanjing Med Univ Dept Thorac Cardiovasc Surg Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008;

    Nanjing Med Univ Dept Thorac Cardiovasc Surg Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008;

    Nanjing Med Univ Dept Nephrol Suzhou Municipal Hosp 242 Guangji Rd Suzhou 215008 Peoples R;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 遗传学;
  • 关键词

    Retroviridae; tPA; Heredity vector; Thrombus; Thrombolysis;

    机译：retroviridae;TPA;遗传矢量;血栓;溶栓;

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