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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、PC
Objective To investigate the effect of 8- Br-c AMP on the expression of tumor suppressor gene HXO-Rb44 in human retinoblastoma and its effect on cell growth. Methods The expressions of p16 INK4m RNA, p21WAF1mRNA, wp5 3m RNA, m p5 3m RNA and Rb m RNA were detected by in situ hybridization and dot blot technique. Immunohistochemistry and protein dot blot were used to detect the expressions of PCNA - IR and p2 1WAF1 - IR, p16 - IR, p Rb - IR, cdk2 - IR and cdk4 - IR. Results The p16 INK4m RNA, p2 1 WAF 1 m RNA, wp5 3m RNA, m p5 3m RNA and Rb m RNA were located in the cytoplasm in human HXO-Rb44 cells, while p16 - IR, p2 1W AF 1 - IR and p Rb - IR were mainly Located in the nucleus. The relative scans of p16 INK4m RNA, p2 1waf1 m RNA, wp5 3m RNA, Rb m RNA and p16 - IR, p2 1 WAF 1 - IR and p Rb - IR were both in the experimental group ( Treated with 8- Br-c AMP) was higher than the respective control group (without 8-Br-cAMP treatment), P <0.05-0.01; whereas mp5 3m RNA, PCNA- IR, cdk2- The relative scans of IR and cdk4-IR were lower in the experimental group than in the control group (P <0.05-0.01). (2) 8-Br-c AMP can up-regulate the expression of tumor suppressor genes, including p16 INK4m RNA, p2 1WAF 1 m RNA, wp5 3m RNA, Rb m RNA and p16, p2 1WAF 1, p Rb protein expression; (3) 8-Br-c AMP can down-regulate mp5 3m RNA, PC