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目的构建携带人肿瘤抑素Tumstatin基因的重组腺相关病毒载体,研究其在乳腺癌细胞MCF-7中的表达。方法以人胚肾293细胞为材料,用RT-PCR方法获取人肿瘤抑素Tumstatin基因,将其克隆入pAAV-MCS载体中形成重组载体pAAV-Tum质粒。用脂质体介导法将重组载体导入乳腺癌细胞株(MCF-7)建立一株稳定的表达Tumstatin基因的乳腺癌细胞株MCF/AAV-Tum,并用RT-PCR检测该细胞系中Tumstatin基因的表达。结果成功构建了pAAV-Tum重组腺相关病毒载体,筛选出乳腺癌细胞株MCF/AAV-Tum,该细胞能表达Tumstatin基因。结论构建的pAAV-Tum重组载体能在MCF-7细胞中表达,为其后的肿瘤抗血管生成治疗研究奠定了基础。
Objective To construct a recombinant adeno-associated virus vector carrying human tumor suppressor Tumstatin gene and study its expression in breast cancer cell line MCF-7. Methods Human embryonic kidney 293 cells were used as material, RT-PCR method was used to obtain human tumor suppressor Tumstatin gene, which was cloned into pAAV-MCS vector to form recombinant vector pAAV-Tum plasmid. A recombinant human breast cancer cell line MCF/AAV-Tum expressing a stable Tumstatin gene was introduced into breast cancer cell line (MCF-7) by liposome-mediated method. The Tumstatin gene in the cell line was detected by RT-PCR. expression. Results The pAAV-Tum recombinant adeno-associated virus vector was successfully constructed and the breast cancer cell line MCF/AAV-Tum was screened out. This cell can express the Tumstatin gene. Conclusion The constructed pAAV-Tum recombinant vector can be expressed in MCF-7 cells, which laid the foundation for the subsequent anti-angiogenesis therapy research.