【摘要】 目的:以重组腺相关病毒(rAAV)为载体介导以HIF-1α为靶点的小干扰RNA,在厌氧培养条件下作用于Mia-PaCa2人胰腺癌细胞,观察其对糖代谢反应关键酶——葡萄糖转运体-1(Glut-1)表达的影响。方法:构建rAAV介导的以HIF-1α为靶基因的RNA干扰表达载体,即rAAV-siHIF。将rAAV-hrGFP、rAAV-siHIF-1α分别转染对数生长的MiaPaCa2人胰腺癌细胞,在乏氧条件下进行培养,应用Real-Time PCR、RT-PCR、Western Blot法观察其对Mia-PaCa2细胞HIF-1α mRNA、蛋白以及Glut-1 mRNA、蛋白表达的影响。结果:Real-Time PCR和Western Blot检测发现,rAAV-siHIF-1α能够抑制MiaPaCa2细胞HIF-1α mRNA和蛋白的表达。RT-PCR和Western Blot检测发现,rAAV-siHIF-1α能抑制MiaPaCa2细胞Glut-1 mRNA和蛋白的表达。结论:rAAV-siHIF-1α能抑制MiaPaCa2人胰腺癌细胞糖代谢关键酶Glut-1表达的作用。更多还原

【Abstract】 Objective:To study the effect of recombinant adeno-associated virus (rAAV) vector carrying siRNA targeting HIF-1α on the expression of Glut-1 in MiaPaCa2 human pancreatic cancer cells under hypoxia primarily. Methods: A cassette encoding siRNA targeting HIF-1α mediated by rAAV was constructed. rAAV-hrGFP and rAAV-siHIF-1α were transfected to exponentially growing MiaPaCa2 cells. Then, the expression of HIF-1α mRNA and protein, the expression of Glut-1 mRNA and protein under hypoxic conditions were observed by Real-Time PCR, RT-PCR and Western Blot. Results: Under hypoxia, rAAV-siHIF-1α could inhibit the expression of HIF-1α mRNA in MiaPaCa2 cells. Moreover, rAAV-si- HIF-1α could inhibit the expression of Glut-1 gene and protein. Conclusion: rAAV-siHIF-1α can inhibit the expression of Glut-1 under hypoxia.更多还原