【摘要】 目的观察反义缺氧诱导因子-1α(HIF-1α)对胰腺癌细胞BxPC-3化疗敏感性的影响。方法实验分组:(1)缺氧条件下(0.5%O2)体外培养4 h,未转染反义HIF-1α质粒的BxPC-3细胞设为缺氧对照组;(2)常氧条件下体外培养,未转染反义HIF-1α质粒的BxPC-3细胞设为常氧对照组;(3)缺氧条件下(0.5%O2)体外培养4 h,稳定转染反义HIF-1α质粒的BxPC-3细胞设为实验组。采用逆转录聚合酶链反应(RT-PCR)和免疫印迹(W estern B lot)检测各组的HIF-1α和survivin表达情况。流式细胞术和MTT比色法检测不同剂量的化疗药物(5-氟尿嘧啶、阿霉素、吉西他宾)对各组的凋亡率和生长抑制率的影响。结果实验组HIF-1α和survivin的表达明显降低(P<0.0 5),与对照组相比,实验组的凋亡率、抑制率与剂量成正比,高剂量引起高抑制(P<0.0 5)。结论反义HIF-1α可能通过阻断survivin的表达而增强胰腺癌对化疗的敏感性。据此可望通过阻断HIF-1α的表达为胰腺癌基因治疗提供一种新途径。更多还原

【Abstract】 Objective To observe the effect of antisense hypoxia inducible factor-1α（HIF-1α） on （chemosensitivity） of human pancreatic cancer cell line BxPC-3 under hypoxia. Methods BxPC-3 cells were divided into 3 groups:（1）BxPC-3 cells were non-transfected with antisense HIF-1α plasmid and exposed to 0.5% O₂ for 4hr（hypoxia control）;（2）normoxic BxPC-3 cells were non-transfected with antisense（HIF-1α） plasmid（normoxia control）;（3）BxPC-3 cells were transfected with antisense HIF-1α plasmid and exposed to 0.5% O₂ for 4hr（experimental group）.Expression of HIF-1α and survivin was detected by RT-PCR and Western Blot.Growth inhibition rates and apoptosis rates of BxPC-3 cells under different（dosages） of chemotherapeutic agents（5-fluorouracil,doxorubicin and gemcitabine） were measured by MTT（colorimetric） assay and flow cytometry （FCM）.Results Expression of HIF-1α was obviously down-regulated and at the same time susvivin expression was markedly down-regulated in experimental group（P<0.05）.Higher dosages(100 mg/L,200 mg/L and 400 mg/L of 5-fluorouracil,0.05 mg/L,0.075 mg/L and 0.1 mg/L of doxorubicin,10^-9 mol/L,10^-8 mol/L and 10^-7 mol/L of gemcitabine) caused a（greater） increase of inhibition in experimental group than in hypoxia control（P<0.05）.Conclusions The results demonstrate that antisense HIF-1α inhibits expression of survivin and enhances chemosensitivity of（human） pancreatic cancer cell BxPC-3.Blocking HIF-1α in pancreatic cancer cells may offer an avenue for gene therapy.更多还原