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

发布时间：2018-10-16 18:52

【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。
[Abstract]:Objective to observe the proliferation and invasion of glioma U251 cells transfected with VEGF small interfering RNA (VEGF-siRNA) and to explore its mechanism. Methods Human glioma U251 cells cultured in vitro were divided into observation group, control group and blank group. The cells were transfected with VEGF-siRNA,siRNA-NC, empty liposome for 48 h. The proliferation of cells in each group was observed by CCK-8 method (expressed as OD_ (490), the invasiveness of cells was observed by Transwell chamber experiment (expressed by the number of transmembrane cells), and the VEGF, matrix metalloproteinase-2 (MMP-2) and matrix metal metalloproteinase (MMP-2) were detected by Western blotting assay. Protease 9 (MMP-9), protein kinase B (AKT), phosphorylated protein kinase B (p-AKT) protein expression. Results after 48 hours of transfection, the relative expression of VEGF protein in the control group and the control group was 0.161 卤0.0400.49 卤0.032 卤0.656 卤0.026 OD490, 0.226 卤0.0180.49 卤0.035 卤0.454 卤0.038, and the number of perforating cells was (100.61 卤16.90), (, 284.11 卤13.1), (, 286.18 卤12.24), respectively. The relative expression of VEGF, OD_ (490) and the number of perforating cells in the observation group were lower than those in the control group and the blank group (all P 0.01). After 48 hours of transfection, the relative expression of MMP-2,MMP-9,p-AKT in observation group was lower than that in control group and blank group (P 0.01), and the relative expression of AKT in observation group was higher than that in control group and blank group (P 0.01). Conclusion the VEGF-siRNA transfected glioma U251 cells have a decreased invasion and proliferation due to the inhibition of MMP-2,MMP-9,AKT,p-AKT expression in U251 cells by transfection of VEGF-siRNA.
【作者单位】：德州市市立医院;
【分类号】：R739.41

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