8-Br-cAMP对人视网膜母细胞瘤HXO-Rb44细胞抑癌基因表达的效应  

THE EFFECT OF 8-Br-cAMP ON EXPRESSION OF ANTIONCOGENES IN HUMAN RETINOBLASTOMA HXO-Rb44 CELLS IN VITRO

作　　者：邓新国[1] 郭雪[2] 吴景兰[3] 郭希让[1] 庞广仁[1] 田小莉[1] 

机构地区：[1]河南省眼科研究所,郑州450003 [2]河南省卫生防疫站,郑州450003 [3]河南医科大学组织学胚胎学教研室,郑州450052

出　　处：《解剖学报》2000年第1期61-64,I012,共5页Acta Anatomica Sinica

基　　金：河南省科委资助! ( 9712 0 0 2 61)

摘　　要：目的　探讨 8- Br- c AMP对人视网膜母细胞瘤之 HXO- Rb44细胞抑癌基因表达的效应及其对细胞生长的影响。　方法　应用原位杂交、RNA斑点印迹技术检测 p16 INK4m RNA、p2 1WAF1 m RNA、wp5 3m RNA、m p5 3m RNA和 Rb m RNA,应用免疫组织化学和蛋白质斑点印迹技术检测 PCNA- IR、p2 1WAF1 - IR、p16 - IR、p Rb- IR、cdk2 - IR和 cdk4- IR。　结果　在人 HXO- Rb44细胞内 p16 INK4m RNA、p2 1WAF 1 m RNA、wp5 3m RNA、m p5 3m RNA及 Rb m RNA位于胞质 ,p16 - IR,p2 1W AF 1 - IR及 p Rb- IR主要位于胞核。在 m RNA和蛋白质斑点印迹标本上 ,p16 INK4m RNA、p2 1waf1 m RNA、wp5 3m RNA、Rb m RNA和 p16 - IR、p2 1WAF 1 - IR及 p Rb- IR的相对扫描数值均是实验组 (经8- Br- c AMP处理 )高于各自的对照组 (未经 8- Br- c AMP处理 ) ,P<0 .0 5～ 0 .0 1;然而 mp5 3m RNA、PCNA - IR、cdk2 - IR和 cdk4- IR的相对扫描数值均是实验组低于对照组 ,P<0 .0 5～ 0 .0 1。　结论　 (1) 8- Br- c AMP可抑制HXO- Rb44细胞的生长增殖 ;(2 ) 8- Br- c AMP可上调抑癌基因的表达 ,包括 p16 INK4m RNA、p2 1WAF 1 m RNA、wp5 3m RNA、Rb m RNA及 p16、p2 1WAF 1 、p Rb的蛋白表达 ;(3) 8- Br- c AMP可下调 mp5 3m RNA。Objective\ To study 8\|Br\|cAMP effect on cell growth and antioncogene expression in human retinoblastoma HXO\|Rb44 cells.\ Methods\ The p16 INK4 mRNA,p21 WAF1 mRNA,w(wild type)p53 mRNA,m(mutant type)p53 mRNA and Rb mRNA were detected by in situ hybridization and RNA dot blot technique.The PCNA,p16,p21 WAF1 ,pRb,cdk2 and cdk4 proteins were detected by immunohistochemistry and protein dot blot technique.\ Results\ The signals of p21 WAF1 mRNA,p16 INK4 mRNA,wp53 mRNA,mp53 mRNA and Rb mRNA were localized in the cytoplasm of HXO\|Rb44 cells.The p16\|IR,p21 WAF1 \|IR and pRb\|IR were predominently localized in the nuclei.In the dot blot the relative scanning value of p16 INK4 mRNA,p21 waf1 mRNA,wp53 mRNA,Rb mRNA and p16\|IR,p21 WAF1 \|IR,pRb\|IR of the exprimental groups(EG,treated with 8\|Br\|cAMP)was higher than that in respective control groups(CG,treated with no 8\|Br\|cAMP), P <0\^05\|0\^01.However,the scanning value of the mp53 mRNA,PCNA\|IR,cdk2\|IR and cdk4\|IR of EG was lower than that in respective control groups, P <0\^05\|0\^01.\ Conclusions\ (1)8\|Br\|cAMP could inhibit human HXO\|Rb44 cell growth and proliferation.(2)8\|Br\|cAMP could up\|regulate antioncogene expression,including mRNA expression of p16 INK4 ,p21 waf1 ,wp53 and Rb and protein expression of p16,p21 WAFl and pRb.(3)8\|Br\|cAMP could down\|regulate expression of mp53 mRNA,PCNA\|IR,cdk2\|IR and cdk4\|IR expression.(4)The results suggest that 8\|Br\|cAMP may decrease human HXO\|Rb44 cell growth via inhibition of cell cycle progressing related gene expression.

关 键 词：视网膜母细胞瘤 抑癌基因 原位杂交 8-BR-CAMP 

分 类 号：R739.702[医药卫生—肿瘤]