【摘要】 背景与目的:有研究表明野生型p53可以诱导RGS16表达,而RGS16可能与胶质瘤的发生有关。本研究旨在探讨RGS16基因转染对大鼠胶质瘤C6细胞生长的影响。方法:构建真核表达载体pIRES2-EGFP-RGS16,脂质体法转染C6细胞,经G418筛选后荧光显微镜观察细胞中增强型绿色荧光蛋白(enhancedgreenfluorscentprotein,EGFP)的表达,RT-PCR证实RGS16的mRNA表达,免疫细胞化学方法检测细胞中RGS16蛋白表达。最后利用流式细胞仪、生长曲线、平板克隆形成等方法研究RGS16基因对胶质瘤细胞周期、生长及增殖的影响。结果:成功构建了稳定表达RGS16和表达空载体的细胞系C6-RGS16、C6-GFP。C6-RGS16、C6-GFP和C6经流式细胞仪检测S期的细胞比例分别为28.5%、18.9%和14.3%(P<0.05);生长曲线表明C6-RGS16生长的速度明显快于C6-GFP及C6,但其平板克隆形成率分别为12%、25%和25%(P<0.05)。结论:RGS16促进C6细胞周期从G1期向S期过渡,加快细胞生长速度,但是并不促进细胞克隆形成。更多还原

【Abstract】 BACKGROUND & OBJECTIVE: Wild type p53 could induceRGS16 mRNA expression, and RGS16 protein is related with carcinogenesisof glioma. This study was designed to investigate the effects of exogenousRGS16 gene transfection on the growth of rat glioma cell line C6.METHODS : A eukaryotic expression plasmid pIRES2-EGFP-RGS16 wasconstructed, and transfected into C6 cells. The cells were selected by G418.The expression of enhanced green fluorescent protein (EGFP) was observedunder fluorescent microscope, and the expression of RGS16 mRNA andprotein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. The effects of RGS16 gene on cell cycleprogression, cell growth, and proliferation of C6 cells were measured by flowcytometry, cell growth curve, and clonogenic assay in plate. RESULTS: C6-RGS16 and C6-GFP cells, which separately expressed RGS16 and pIRES2-EGFP vector, were constructed successfully. S phase proportion wassignificantly higher in C6-RGS16 cells than in C6-GFP and C6 cells (28.5%vs. 18.9% and 14.3%, P<0.05) ; C6-RGS16 cells grew faster than C6-GFPand C6 cells. The clone formation rate was significantly lower in C6-RGS16cells than in C6-GFP and C6 cells (12% vs. 25% and 25% , P <0.05).CONCLUSION: Exogenous RGS16 gene could promote cell cycle progressionof glioma C6 cells from G1 phase to S phase, and accelerate cell growth,but canSt increase the clone formation.更多还原