腺病毒介导PTEN基因抗脑胶质瘤作用的实验研究

目的 :探讨 10号染色体上缺失的磷酸酶和张力蛋白同源物 (phosphataseandtensinhomologydeletedonchromo someten ,PTEN )对人脑胶质瘤细胞的抗增殖作用 ,包括细胞周期改变、抗血管生成的作用。方法 :用含PTEN基因的重组腺病毒载体 ,体外转染人脑胶质瘤细胞U87,用RT PCR、Westernblot检测基因及蛋白的表达。通过细胞生长试验、流式细胞仪分析技术检测转染细胞的增殖、细胞周期的改变 ,用MTT法检测PTEN对人脐静脉内皮细胞生长的影响 ,体内实验检测其致瘤能力。结果 :RT PCR、Westernblot检测转染AdPTEN病毒后的U87细胞PTEN表达由阴性转阳性 ,转染后对U87细胞的体外生长有明显抑制作用 ,使细胞出现G0 ～G1期阻滞 ,并抑制其体内肿瘤生长。含PTEN的上清对内皮细胞生长有明显抑制作用。体内实验表明 ,AdPTEN治疗组可明显抑制肿瘤的生长。结论 :重组腺病毒载体介导的PTEN基因在体内外对人脑胶质瘤细胞U87的生长有抑制作用 ,并抑制肿瘤血管生成 ,具有显著的抗胶质瘤作用

Experiment of adenovirus-mediated PTEN gene therapy for malignant glioma

OBJECTIVE:To evaluate the effects of adeno virus-mediated in vitro gene transfer of PTEN in cell proliferation and angiogenesis of malignant glioma cells U87.METHODS:PTEN expression was examined by RT-PCR and Western blotting.The growth rate of U87 and ECV304 cell were assayed by using MTT assay.The cell cycle was measured by flow cytometry.The growth of tumors were observed by using nude mice model.RESULTS:PTEN expressed stably in glioma cell.Ad-PTEN inhibited the cell growth and proliferation effectively in vitro and make tumor regression in vivo.The progression of cell cycle was arrested from G 1 to S phase.CONCLUSIONS:Adenovirus-mediated PTEN gene therapy for malignant glioma would have wide application in the future.