反义缺氧诱导因子-1α对胰腺癌细胞BxPC-3化疗敏感性的影响  

Effect of gene transfer of antisense hypoxia inducible factor-1α on chemosensitivity of human pancreatic cancer cell line BxPC-3

作　　者：常青[1] 秦仁义[1] 高军[1] 冯延平[1] 黄涛[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院胆胰外科,湖北武汉430030

出　　处：《中国普通外科杂志》2006年第6期423-427,共5页China Journal of General Surgery

基　　金：国家自然科学基金资助项目(30471693)

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。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-let plasmid （normoxia control ） ; （3）BxPC-3 cells were transfected with antisense HIF-1 et plasmid and exposed to 0. 5 % O2 for 4 hr （ 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, doxornbicin and gemcitabine ） were measured by MTT colorimetric assay and flow cytometry （ FCM ）. Results Expression of HIF- 1 et was obviously down- regulated and at the same time survivin 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 doxornbicin, 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-1 et inhibits expression of survivin and enhances chemosensitivity of human pancreatic cancer cell BxPC-3. Blocking HIF-1 et in pancreatic cancer cells may offer an avenue for gene therapy.

关 键 词：胰腺肿瘤/药物疗法 反义缺氧诱导因子-1α 化疗敏感性 基因疗法 

分 类 号：R735.9[医药卫生—肿瘤] R459.9[医药卫生—临床医学]