轉(zhuǎn)染VEGF小干擾RNA的腦膠質(zhì)瘤細胞U251增殖、侵襲能力變化及其機制
發(fā)布時間：2018-10-16 18:52
【摘要】：目的觀察轉(zhuǎn)染VEGF小干擾RNA(VEGF-siRNA)的腦膠質(zhì)瘤U251細胞增殖、侵襲能力的變化,并探討及其機制。方法將體外培養(yǎng)的人腦膠質(zhì)瘤U251細胞分為觀察組、對照組和空白組,分別用VEGF-siRNA、siRNA-NC、空脂質(zhì)體轉(zhuǎn)染48 h后,用CCK-8法觀察各組細胞增殖情況(以O(shè)D_(490)表示),用Transwell小室實驗觀察各組細胞侵襲能力(以穿膜細胞數(shù)表示),用Western blotting法檢測各組細胞中的VEGF、基質(zhì)金屬蛋白酶2(MMP-2)、基質(zhì)金屬蛋白酶9(MMP-9)、蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)蛋白表達。結(jié)果轉(zhuǎn)染48 h后觀察組、對照組、空白組細胞VEGF蛋白相對表達量分別為0.161±0.040、0.649±0.032、0.656±0.026;OD_(490)分別為0.226±0.018、0.449±0.035、0.454±0.038;穿膜細胞數(shù)分別為(100.61±16.90)、(284.11±13.1)、(286.18±12.24)個。觀察組細胞VEGF相對表達量、OD_(490)、穿膜細胞數(shù)均低于對照組和空白組(P均0.01)。轉(zhuǎn)染48 h后觀察組細胞MMP-2、MMP-9、p-AKT相對表達量均低于對照組和空白組(P均0.01);觀察組細胞AKT相對表達量與對照組和空白組相比,P均0.01。結(jié)論轉(zhuǎn)染VEGF-siRNA的腦膠質(zhì)瘤U251細胞增殖、侵襲能力下降,其機制是由于轉(zhuǎn)染VEGF-siRNA抑制U251細胞MMP-2、MMP-9、AKT、p-AKT表達。
[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 鹵0.0400.49 鹵0.032 鹵0.656 鹵0.026 OD490, 0.226 鹵0.0180.49 鹵0.035 鹵0.454 鹵0.038, and the number of perforating cells was (100.61 鹵16.90), (, 284.11 鹵13.1), (, 286.18 鹵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.
【作者單位】： 德州市市立醫(yī)院;
【分類號】：R739.41
本文編號：2275312
[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 鹵0.0400.49 鹵0.032 鹵0.656 鹵0.026 OD490, 0.226 鹵0.0180.49 鹵0.035 鹵0.454 鹵0.038, and the number of perforating cells was (100.61 鹵16.90), (, 284.11 鹵13.1), (, 286.18 鹵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.
【作者單位】： 德州市市立醫(yī)院;
【分類號】：R739.41
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