CAJ | 학술논문

构建重组质粒pcDNA4/HisA-LMO3,转染C8-D1A胶质细胞,对照组为pcDNA4/HisA空载体质粒转染组和未转染组,MTT观察各组细胞的体外生长情况,流式细胞术(FCM)测定各组细胞的细胞周期和凋亡细胞百分比,Western-blot检测LMO3转染后凋亡相关蛋白的表达变化,从而观察LMO3基因对C8-D1A胶质细胞体外生长的影响.RT-PCR、Western印迹显示pcDNA4/HisA-LMO3转染组LMO3mRNA及LMO3蛋白表达水平明显高于对照组;与对照组细胞相比,转染组细胞的增殖能力明显高于空载体对照组及C8细胞(P<0.01),S期细胞增加,G_0/G_1期细胞减少,C8细胞转染LMO3后可以促进C8细胞从G_0/G_1期进入S期,从而促进细胞的增殖.
구건중조질립pcDNA4/HisA-LMO3,전염C8-D1A효질세포,대조조위pcDNA4/HisA공재체질립전염조화미전염조,MTT관찰각조세포적체외생장정황,류식세포술(FCM)측정각조세포적세포주기화조망세포백분비,Western-blot검측LMO3전염후조망상관단백적표체변화,종이관찰LMO3기인대C8-D1A효질세포체외생장적영향.RT-PCR、Western인적현시pcDNA4/HisA-LMO3전염조LMO3mRNA급LMO3단백표체수평명현고우대조조;여대조조세포상비,전염조세포적증식능력명현고우공재체대조조급C8세포(P<0.01),S기세포증가,G_0/G_1기세포감소,C8세포전염LMO3후가이촉진C8세포종G_0/G_1기진입S기,종이촉진세포적증식.
C8 glia cells were cultured in DMEM with 10% FBS. Recombinant plasmid pcDNA4/HisA-LMO3 was constructed and transfected into the C8 cells with the lipofectamine 2000. A stable C8 cell line overexpressing LMO3 was established to observe the direct effect of LMO3 on C8 cells. Parental C8 cells and parental C8 cells stably transfected with blank vector pcDNA4/HisA were used as control groups. The cell growth was determined by MTT method and flow cytometry (FCM) was used to analyze the cell cycle in each group. The stably transfected cell line C8-LMO3 was confirmed by RT-PCR and Western blotting. After transfection with LMO3, C8 cells showed significant increase in cell proliferation when compared with the control groups. The percentage of S phase cells increased, whereas G_0 / G_1 phase cells decreased. Overexpression of LMO3 can stimulate proliferation by promoting glia cell into cell cycle.