I型免疫缺陷病毒蛋白R降低U251细胞恶性生物学行为的研究  

HIV-1 viral protein R decreases malignant behavior of glioma cell U251

作　　者：张飚[1] 张述升[2] 冯学泉[3] 徐新女[3] 王金环[2] 

机构地区：[1]天津市环湖医院检验科,300060 [2]天津市神经外科研究所 [3]天津市第一中心医院

出　　处：《中华神经外科杂志》2014年第5期463-467,共5页Chinese Journal of Neurosurgery

基　　金：天津市应用基础及前沿技术研究计划（11JCYBJC12100）;天津市抗癌重大专项计划（12ZDZSY17700）;天津市卫生局科技计划项目（11KG115）;国家临床重点专科建设专项基金资助项目（2011-327）

摘　　要：目的探讨重组腺病毒介导的I型人类免疫缺陷病毒蛋白R（Vpr）转染对胶质瘤U251细胞TGFl3RIll表达及恶性生物学行为的影响。方法将U251细胞分为正常对照组，空载体组和实验组进行细胞培养，空载体组和实验组按感染复数（multiplicityofinfection，MOI）=100分别进行空载腺病毒（Ad—GFP）和含有Vpr基因的重组腺病毒（Ad—Vpr）转染，转染后用Westernblot检测Vpr、TGFl3RIU、MMP-2和MMP-9表达水平，Transwell、划痕实验检测细胞迁移和侵袭能力。结果实验组U251细胞经Ad-Vpr转染后，可见Vpr蛋白表达，同时TGFf3RIU蛋白表达升高，正常对照组、Ad-GFP组、Ad-Vpr组TGFβRⅢ与B—actin密度灰度比值分别为0．71±0．04，0．83±0．09和1．13±0．16，三组间差异有统计学意义（P〈0．05）。MMP-2和MMP-9蛋白表达降低，Ad．Vpr组MMP-2和MMP-9与13-actin密度灰度比值分别为0．51±0．11和0．43±0．08，对照组为1．11±0．11和1．06±0．18，Ad-GFP组为1．23±0．05和1．16±0．12，与对照组和Ad—GFP组相比，Ad—Vpr组MMP-2和MMP-9表达明显降低（P〈0．05）。Transwell体外侵袭实验结果显示，对照组、Ad—GFP组和Ad-Vpr组平均视野的穿膜细胞数分别为（123．12±10．82）个、（118．89±12．65）个和（80．64±7．72）个，与对照组和Ad—GFP组比较，Ad-Vpr转染组穿膜细胞数明显减少，差异具有统计学意义（P〈0．05）。结论Ad-Vpr可能通过上调TGF±RⅢ降低胶质瘤U251细胞的迁移和侵袭能力。Objective To study the effect of recombinant adenovirus mediated transfection of human immunodeficiency virus type 1 viral protein R （Vpr） on TGFβR Ⅲ expression and malignant behavior of glioma cell U251. Methods U251 ceils were divided into the control group, mock group and experiment group, the mock and experiment group were transfected with Ad-GFP and Ad-Vpr, respectively at a multiplicity of infection （MOI） of 100. The expression of Vpr, TGFβR Ⅲ, MMP-2 and MMP-9 were detected by western blot, the malignant biological behavior changes of U251 cells transfected by Ad-Vpr were evaluated by scratch assay and transwell assay. Results Western blot showed that the Vpr protein could be expressed after Ad-Vpr transfection, meanwhile, the expression of TGFβR Ⅲ was up regulated by Ad-Vpr transfection, Relative density values of TGFβRⅢ compared to β-actin in control, Ad-GFP and Ad-Vpr group were0. 71 ±0. 04,0. 83± 0. 09 and 1.13 ±0. 16, respectively, there were significant different in three groups（ P 〈 0. 05 ）. While the expression of MMP-2 and MMP-2 were down regnlated by Ad-Vpr transfection. Relative density values of MMP-2 and MMP-9 compared to 13-actin were 0. 51 ± 0. 11 and 0. 43 ±0. 08 for Ad-Vpr group, 1.11 ±0. 11 and 1.06±0. 18 for control group and 1.23 s0.05 and 1.16 ± 0. 12 for Ad-GFP group, respectively. The expression of MMP-2 and MMP-9 were significantly decreased comparing to control and Ad-GFP group （ P 〈 0. 05 ）. Transwell assay results showed that the average ceils across the membrane were 123.12 ± 10. 82, 118.89 ±12. 65 and 80. 64 ± 7.72 in control, Ad-GFP and Ad-Vpr group, respectively. Compared with control and the Ad-GFP group, the cells across the membrane in Ad-Vpr group were significantly reduced （ P 〈 0. 05 ）. Conclusions Vpr could decrease migratory and invasive behavior of glioma U251 cells probably by upregnlating TGFI3Rm expression.

关 键 词：神经胶质瘤 免疫缺陷病毒蛋白R 转化生长因子β受体Ⅲ 

分 类 号：R739.41[医药卫生—肿瘤]