转染VEGF小干扰RNA的脑胶质瘤细胞U251增殖、侵袭能力变化及其机制  

Changes of proliferation and invasion of glioma U251 cells transfected with VEGF small interfering RNA

作　　者：侯保辉 

机构地区：[1]德州市市立医院,山东德州253012

出　　处：《山东医药》2017年第19期29-32,共4页Shandong Medical Journal

摘　　要：目的观察转染VEGF小干扰RNA(VEGF-siRNA)的脑胶质瘤U251细胞增殖、侵袭能力的变化,并探讨及其机制。方法将体外培养的人脑胶质瘤U251细胞分为观察组、对照组和空白组,分别用VEGF-siRNA、siRNA-NC、空脂质体转染48 h后,用CCK-8法观察各组细胞增殖情况(以OD_(490)表示),用Transwell小室实验观察各组细胞侵袭能力(以穿膜细胞数表示),用Western blotting法检测各组细胞中的VEGF、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)蛋白表达。结果转染48 h后观察组、对照组、空白组细胞VEGF蛋白相对表达量分别为0.161±0.040、0.649±0.032、0.656±0.026;OD_(490)分别为0.226±0.018、0.449±0.035、0.454±0.038;穿膜细胞数分别为(100.61±16.90)、(284.11±13.1)、(286.18±12.24)个。观察组细胞VEGF相对表达量、OD_(490)、穿膜细胞数均低于对照组和空白组(P均<0.01)。转染48 h后观察组细胞MMP-2、MMP-9、p-AKT相对表达量均低于对照组和空白组(P均<0.01);观察组细胞AKT相对表达量与对照组和空白组相比,P均>0.01。结论转染VEGF-siRNA的脑胶质瘤U251细胞增殖、侵袭能力下降,其机制是由于转染VEGF-siRNA抑制U251细胞MMP-2、MMP-9、AKT、p-AKT表达。Objective To investigate the changes of proliferation and invasion of glioma U251 cells transfected with vascular endothelial growth factor （VEGF） siRNA （VEGF-siRNA） and its mechanism. Methods Human glioma U251 cells were divided into the observation group, control group, and blank group which were transfected with VEGF-siRNA, siRNA-NC, and empty plasmid for 48 h, respectively. CCK-8 method was used to observe the proliferation of cells in each group （expressed in OD_490）, Transwell chamber was used to observe the invasion ability of each group （ expressed as a number of transmembrane cells）, Western blotting was used to detect the protein expression of matrix metalloproteinase 2 （ MMP-2）, matrix metalloproteinase 9 （ MMP-9 ）, protein kinase B （AKT） and phosphorylated protein kinase B （ p-AKT）. Results At 48 h after transfection, the. observation group had significantly lower VEGF protein expression levels, OD_490 and smaller number of transmembrane cells than the control group and blank group （ VEGF protein expression ： 0. 161 ± 0.040 vs 0.649 ± 0. 032 and 0. 656 ± 0.026, OD_490 ： 0.226 ± 0. 018 vs 0.449 ±0. 035 and 0.454 ± 0. 038, the number of transmernbrane cells ： 100.61 ± 16.90 vs 284.11 ± 13.1 and 286.18 ±12.24 ; all P 〈 0.01 ）. The expression of MMP- 2, MMP-9 and p-AKT in the observation group was lower than that of the control group and the blank group at 48 h after transfection （all P 〈0.01 ）. No significant difference was found in the expression of AKT between the observation group and the control group, the blank group, all P 〉 0.01. Conclusion The proliferation and invasion abilities of glioma U251 cells transfected with VEGF-siRNA decrease by inhibiting the expression of MMP-2, MMP-9, AKT, and p-AKT.

关 键 词：微小RNA 血管内皮生长因子 脑胶质瘤 细胞增殖 肿瘤侵袭 基质金属蛋白酶 蛋白激酶B 

分 类 号：R739.41[医药卫生—肿瘤]