人Tumstatin基因真核表达载体的构建及其活性研究

The Construction of Recombinant AAV Vector Mediated Tumstatin Gene and the Detection of Its Activity

目的:构建携带人肿瘤抑素Tumstatin基因的重组腺相关病毒载体,研究其人脐静脉内皮细胞(HUVECs)增殖的影响。方法:构建重组载体pAAV-Tum质粒,并建立一株稳定表达Tumstatin基因的HUVECs细胞株。用RT-PCR检测该细胞系中Tumstatin基因的表达,MTT法检测该细胞增殖情况。结果:成功构建了pAAV-Tum重组腺相关病毒载体,该重组载体能在HUVECs细胞中表达,其表达产物能抑制该细胞增殖。结论:构建的pAAV-Tum重组载体为其后的肿瘤抗血管生成治疗研究奠定了基础。

Objective To construct the recombinant adeno-associated virus(rAAV) vector containing tumstatin gene and to detect its effect on the proliferation ability of HUVECs cells.Methods The pAAV-Tum recombinant plasmid was obtained,and HUVECs cell line stably expressing tumstatin gene was established.Then the mRNA level of pAAV-Tum was detected by RT-PCR and the proliferation ability of transfected cell line was measured by MTT assay.Results Recombinant AAV vector,which contained Tumstatin gene,was constructed successfully.The pAAV-Tum recombinant plasmid can express in HUVECs cell and its expression product can suppress the proliferation of HUVECs cell.Conclusion The recombinant vector of pAAV-Tum,will lead a foundation for its antiangiogenesis theraphy of tumor.