人肿瘤抑素基因真核表达载体的构建及其活性研究

Study on the construction of recombinant AAV vector mediated tumstatin gene and 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,研究其对人脐静脉内皮细胞(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HU—VECs细胞株。用逆转录聚合酶链反应检测该细胞系中Tumstatin基因的表达,四甲基唑蓝法检测该细胞增殖情况。结果构建的pAAV—Turn重组腺相关病毒载体能在HUVECs细胞中表达,其表达产物能抑制该细胞增殖(0.335±0.011)。结论成功构建了pAAV—Turn重组载体,构建的重组载体能有效表达,为其后的抗肿瘤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 Turn recombinant plasmid was obtained,and HUVECs cell line stably expressing Tumstatin gene Was established.Then the mRNA level of pAAV Turn was detected by RT-PCR and the proliferation ability of transfected cell line was measured by MTTassay.Results The pAAV Turn recombinant plasmid could express in HUVECs cell and its expression product could suppress the proliferation of HUVECs cell.Conclusion Recombinant AAV vector,which contained Tumstatin gene,was constructed successfully,and can express efficiently.