中文摘要：

目的 观察LY294002联合重组腺病毒Ad-第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因（PTEN）对胶质瘤LN229细胞株增殖和侵袭的影响.方法 应用噻唑蓝（MTT）法、流式细胞术、划痕实验和Transwell细胞侵袭实验评价LY294002联合Ad-PTEN对LN229细胞增殖、侵袭的抑制作用;Western blot检测PTEN/磷酸肌醇3激酶（PI3K）/蛋白激酶B（Akt）信号通路中蛋白表达的变化.结果 与二甲基亚砜（DMSO）组、空载组和LY294002组比较,联合组细胞生长明显受到抑制,第6天细胞增殖率仅为38.68％;细胞周期分析结果表明,联合组细胞G0/G1期比例为75.49％,高于DMSO组、空载组和LY294002组的51.62％、50.31％、66.35％,差异有统计学意义（P＜0.05）;Transwell法分析结果显示,联合组穿过小室的细胞数明显减少,仅为（15.58±1.45）个,低于DMSO组、空载组和LY294002组的（27.63±4.36）、（46.67 ±3.61）、（49.28±5.37）个,差异有统计学意义（P＜0.05）.Western blot检测DMSO组、空载组、LY294002组和联合组PTEN蛋白相对表达量分别为0.13 ±0.04、0.26±0.03、0.52±0.07和0.83±0.16,磷酸化蛋白激酶B（p-Akt）蛋白相对表达量分别为1.30±0.07、1.73±0.07、0.87 ±0.17和0.35±0.08,增殖细胞核抗原（PCNA）蛋白相对表达量为2.72±0.02、3.13±0.06、0.63 ±0.13和0.38±0.09,细胞周期素D1（Cyclin D1）蛋白相对表达量为1.72±0.09、1.18 ±0.15、0.49±0.11和0.18 ±0.14,磷酸化黏着斑激酶（p-FAK）蛋白相对表达量为0.95±0.14、1.09 ±0.16、1.29±0.08和0.65 ±0.11,基质金属蛋白酶-2（MMP-2）蛋白相对表达量为0.73 ±0.19、0.87±0.12、0.61±0.10和0.23±0.08,表明联合组PTEN蛋白表达显著上调,p-Akt、PCNA、Cyclin D1、MMP-2、p-FAK表达显著下调,差异均有统计学意义（P＜0.05）.结论 LY294002联合Ad-PTEN通过调控PI3K/Akt信号通路对胶质瘤LN229细胞株的增殖和侵袭具有联合抑制作用。

英文摘要：

Objective To investigate the effect of combinatorial therapy with adenoviral-mediated phosphatase and tensin homologue deleted on chromosometen （PTEN） and a phosphoinositide 3-kinase （PI3K） inhibitor LY294002 suppresses malignant glioma cell LN229 proliferation and invasion by regulating the PTEN/PI3K/protein kinase B （Akt） signaling pathway.Methods Ad-PTEN and LY294002 were transfected into LN229 malignant glioma cells,and they were used to detect the proliferative activity of the LN229 cells by methyl thiazol tetrazolium （MTF） and flow cytometry.The invasion ability of LN229 cells was examined by Transwell test.Western blotting analysis was used to detect the protein expression in PTEN/PI3K/Akt signal pathway.Results Compared with Ad-vector,dmethyl sulfoxide （DMSO） and LY294002 group,the MTT assay showed that the cell growth was significantly inhibited in the combination group and the cell proliferation rate was only 38.68% on the six day.The cell cycle analysis showed that the DNA contents of cell G0/G1 phase in the combination group was 75.49% . It was higher than 51.62%,50.31% and 66.35% in the Ad-vector,DMSO and LY294002 group.There were significant differences （P 〈0.05）.The Transwell assay decreased obviously,and there were only 15.58 ± 1.45.It was lower than 27.63 ±4.36,46.67 ±3.61 and 49.28 ±5.37 in the Ad-vector,DMSO and LY294002 group.There were significant differences （P 〈 0.05）.Western blotting assay showed that the relative expression quantities of PTEN were 0.13 ± 0.04,0.26 ± 0.03,0.52 ± 0.07 and 0.83 ± 0.16,respectively;and the relative expression quantities of phosphorylated protein kinase B （p-Akt） were 1.30 ± 0.07,1.73 ±0.07,0.87 ±0.17 and 0.35 ±0.08,respectively;and the relative expression quantities of proliferating cell nuclear antigen （PCNA） were 2.72 ±0.02,3.13 ±0.06,0.63 ±0.13 and 0.38 ±0.09,respectively;and the relative expression quantities of Cyclin D1 were 1.72 ± 0.09,1.18 ± 0.15,0.49 ± 0.11 and 0.18 ±0.14,respectively;and th