CAJ | 학술논문

目的：探讨靶向上调和下调谷胱甘肽过氧化物酶1（GPx1）对人胶质母细胞瘤细胞株（U87MG、U118MG）增殖、迁移和侵袭能力的影响。方法：合成针对GPx1的siRNA和构建、鉴定pcDNA3.1-GPx1重组质粒，分别用 LipofectamineTM 2000脂质体瞬时转染人胶质母细胞瘤细胞株 U87MG 和 U118MG。 Real-time PCR 检测U87MG和U118MG中GPx1 mRNA表达。 Western blotting法检测转染后GPx1蛋白表达，用MTS检测法和Transwell实验分别检测转染后细胞的活力、迁移及侵袭能力的变化。结果：siRNA干扰U87MG细胞后在24 h、48 h和72 h对细胞活力的抑制率分别为25.9％、35.7％和34.8％，较对照组明显降低（P＜0.05）；质粒转染U118MG细胞后在24 h、48 h和72 h对细胞活力的促进率分别为22.7％、45.8％和39.8％，较对照组明显增加（ P＜0.05）；Transwell结果显示经siRNA干扰，U87MG细胞迁移抑制率为41.6％±8.2％，细胞侵袭抑制率为40.4％±10.1％，经siRNA干扰的细胞迁移和侵袭能力明显下降（ P＜0.05）；质粒转染后细胞迁移促进率为55.8％±9.8％，细胞侵袭促进率为60.8％±9.2％，经质粒转染的细胞的迁移和侵袭能力显著增强（ P＜0.05）。结论： siRNA下调GPx1表达可抑制人胶质母细胞瘤细胞株U87MG的生长、迁移和侵袭；相反，质粒上调GPx1表达可促进人胶质母细胞瘤细胞株U118MG的生长、迁移和侵袭。
목적：탐토파향상조화하조곡광감태과양화물매1（GPx1）대인효질모세포류세포주（U87MG、U118MG）증식、천이화침습능력적영향。방법：합성침대GPx1적siRNA화구건、감정pcDNA3.1-GPx1중조질립，분별용 LipofectamineTM 2000지질체순시전염인효질모세포류세포주 U87MG 화 U118MG。 Real-time PCR 검측U87MG화U118MG중GPx1 mRNA표체。 Western blotting법검측전염후GPx1단백표체，용MTS검측법화Transwell실험분별검측전염후세포적활력、천이급침습능력적변화。결과：siRNA간우U87MG세포후재24 h、48 h화72 h대세포활력적억제솔분별위25.9％、35.7％화34.8％，교대조조명현강저（P＜0.05）；질립전염U118MG세포후재24 h、48 h화72 h대세포활력적촉진솔분별위22.7％、45.8％화39.8％，교대조조명현증가（ P＜0.05）；Transwell결과현시경siRNA간우，U87MG세포천이억제솔위41.6％±8.2％，세포침습억제솔위40.4％±10.1％，경siRNA간우적세포천이화침습능력명현하강（ P＜0.05）；질립전염후세포천이촉진솔위55.8％±9.8％，세포침습촉진솔위60.8％±9.2％，경질립전염적세포적천이화침습능력현저증강（ P＜0.05）。결론： siRNA하조GPx1표체가억제인효질모세포류세포주U87MG적생장、천이화침습；상반，질립상조GPx1표체가촉진인효질모세포류세포주U118MG적생장、천이화침습。
AIM: To verify the role of enhancing or suppressing the expression of glutathione peroxidase 1 (GPx1) in the growth, migration and invasion of glioblastoma multiforme cell lines U87MG and U118MG.METHODS:U87MG and U118MG cell lines were transfected with the vector containing specific siRNA or pcDNA3.1 recombinant plas-mid both targeting GPx1.The mRNA and protein expression levels of GPx1 were detected by real-time PCR and Western blotting.MTS assay was applied for determining the cell activity.The abilities of migration and invasion were examined by Transwell assay.RESULTS:Compared with blank control group and negative group, the inhibitory rate of the cell activity in U87MG cells in siRNA group was significantly reduced by 25.9%, 35.7%and 34.8%at 24 h, 48 h and 72 h, respec-tively (P<0.05).In contrast, the cell activity of U118MG cells in pcDNA3.1-GPx1 group was significantly increased by 22.7%, 45.8%and 39.8%at 24 h, 48 h and 72 h, respectively ( P<0.05) .In siRNA group, the inhibitory rate of mi-gration in U87MG cells was 41.6%±8.2%and the invasion was 41.6%±8.2%compared with blank control group and negative group (P<0.05).The cell migration and invasion rates of the U118MG cells in pcDNA-GPx1 group were in-creased by 55.8%±9.8% and 60.8% ±9.2%, respectively, compared with blank control group and negative group (P<0.05).CONCLUSION:The down-regulation of GPx1 by specific siRNA reduces the capability of cell growth, mi-gration and invasion of U87MG cells, while up-regulation of GPx1 by pcDNA3.1-GPx1 increases the capability of cell growth, migration and invasion of U118MG cells.