8-Br-cAMP对人视网膜母细胞瘤细胞系癌基因和抑癌基因作用的研究  

Effects of 8 Br cAMP on expression of oncogenes and antioncogenes in human retinoblastoma lines

作　　者：邓新国[1] 郭希让[1] 吴景兰[2] 庞广仁[1] 田小莉[1] 

机构地区：[1]河南省眼科研究所,河南郑州450003 [2]河南医科大学组织胚胎教研室,河南郑州450052

出　　处：《河南医学研究》2000年第1期28-29,共2页Henan Medical Research

摘　　要：目的 :为探讨 8 Br cAMP对人视网膜母细胞瘤之HXO Rb44细胞系的生长作用机理 ,我们观察了 8 Br cAMP对该细胞的癌基因和抑癌基因表达的体外效应。方法 :将体外培养的HXO Rb44细胞系等分为两组 ,一组为加 2× 10 5mol/L 8 Br cAMP的实验组 (EG) ,另一组为不加 8 Br cAMP的对照组 (CG)。将两组的细胞悬液滴于硝酸纤维素膜 (NCM) ,应用完整细胞RNA斑点印迹和蛋白质斑点印迹免疫组化技术分别检测c fos ,N mycp2 1ras ,野生型 (w)p5 3 ,突变型 (m)p5 3 ,Rb ,p16的mRNA和PCNA的蛋白表达。结果 :wp5 3 ,Rb ,p16及p2 1waf1的mRNA信号为EG >CG(P <0 0 5～ 0 0 1) ;c fos ,N myc ,p2 1ras的mRNA信号和PCNA IR则为EG <CG(P <0 0 5～ 0 0 1)。结论 :8 Br cAMP可上调wp5 3 ,Rb ,p16及p2 1waf1抑癌基因的表达 ,而下调c fos ,N myc ,H ras癌基因的表达 ,发挥抑制HXO Rb44细胞生长的作用 ,这提示 8 Br cAMP对人视网膜母细胞瘤可具有治疗作用。Objective: It is known that 8 Br cAMP is one of selective binding site analogues for cAMP RIIa to affect cell growth through regulation of gene expression;c fos,N myc and H ras belong to oncogenes.p53,Rb,p16 and p21waf1 are antioncogenes and PCNA reflects the cell proliferation.Oncogenes and antioncogenes affect cell growth through control of cell cycle.The aim of this study is to investigate the 8 Br cAMP effect on expression of oncogenes and antioncogenes in human HXO Rb44 cells in vitro.Methods: The cultured HXO Rb44 cells were devided into two aliquots.2×10 -5 mol/L of 8 Br cAMP was added into one aliquot as the experiment group(EG),the another aliquot without 8 Br cAMP as the control group(CG).After 24 hours,the celll suspension was dropped onto the nitrocellulose membrane(NCM).The c fos mRNA,N myc mRNA,p21ras mRNA,Rb mRNA,wild type (w) p53 mRNA,mutant type (m)p53 mRNA,p16 mRNA and p21 waf1 mRNA were detected with respective biotin labeled cDNA probes by intact cell RNA dot blot.Immunoreactivity (IR)of PCNA was detected with spicific antibodies by protein dot blot.Results: The mRNA dot blot signals of c fos,N myc,p21ras,mp53 and PCNA IR in EG were weaker than those in CG(P<0.05～0.01).While,the mRNA signals of Rb,wp53 p16 and p21waf1 in EG were stronger than those in CG( P <0.05～0.01).Conclusion: 8 Br cAMP could up regulate expression of antioncogenes including wp53,Rb,p16 and p21waf1 and down regulate expression of oncogenes,including c fos,N myc and p21ras and PCNA expression,while inhibit HXO Rb44 cell growth.The results suggest that 8 Br cAMP may contribute to treatment of human retinoblastoma.

关 键 词：视网膜母细胞瘤 癌基因 抗癌基因 斑点印迹技术 

分 类 号：R739.7[医药卫生—肿瘤]