摘要目的 观察乏氧环境下PTEN和KAI1基因双转染人胰腺癌AsPC1细胞后对其增殖、迁移的影响.方法 应用我们前期构建的PTEN和KAI1基因过表达载体同时转染乏氧环境培养的人胰腺癌AsPC1细胞,采用蛋白质印迹法检测PTEN和KAI1蛋白的表达,四甲基偶氮唑蓝(MTT)法检测细胞的增殖,克隆形成实验观察肿瘤细胞形成集落能力,Transwell小室实验观察细胞的迁移能力.结果 双基因转染后,乏氧培养的AsPC1细胞的PTEN、KAI1蛋白表达量较空载体转染组细胞增加2.05倍及1.51倍；细胞增殖显著减缓(0.5比0.8,P=0.000)；克隆形成数显著下降[(41.67±5.03)个比(86.00±7.81)个,P=0.017)]；细胞迁移能力明显减弱(0.68±0.05比1.23±0.03,P=0.003).结论 乏氧条件下PTEN、KAI1基因双转染AsPC1细胞后能够抑制其增殖、迁移能力,基因联合治疗胰腺癌可能具有潜在的应用价值.

abstractsObjective To investigate the effects of PTEN and KAI1 double gene transfection on proliferation,metastasis in AsPC1 pancreatic cancer cells under hypoxic condition.Methods Recombinant vectors that over expressing PTEN and KAI1 protein which was established previously were double transfected into hypoxic AsPC1 cells.Western blot was performed to measure the expression level of PTEN and KAI1.Then,cell proliferation was detected by MTT,and colony forming assay were used to test the ability of tumor cells forming colonies,and transwell assay was used to evaluate metastatic function.Results After double gene trnsfection,both PTEN and KAI1 protein expression was significantly up-regulated,which was 2.05 and 1.5 1 folds higher than that of empty vector group; and cell proliferation of hypoxic AsPC1 cells was suppressed significantly (0.5 vs 0.8,P =0.00031 ),number of colony formation was significantly decreased [ (41.67 ± 5.03) vs (86.00 ± 7.81 ),P =0.017) ] ; the capacity of metastasis was significantly decreased (0.68 ± 0.05 vs 1.23 ± 0.03,P =0.0025 ).Conclusions Double gene transfection of PTEN and KAI1 could inhibit proliferation and metastatic activity of hypoxic AsPC1 cells,which might indicate that combined gene therapy may play a role in the treatment of pancreatic cancer.