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目的 探讨8BrcAMP对人视网膜母细胞瘤HXORb44细胞抑癌基因表达的效应。方法 应用RNA斑点印迹技术检测细胞p16、p21wafl、wp53、mp53和Rb的mRNA,应用免疫组化斑点印迹技术检测细胞P16、P21wafl、PRb、cdk2、cdk4和PCNA蛋白质表达的免疫反应性(IR)。结果 在斑点印迹标本上,人HXORb44细胞p16、p21wafl、wp53及Rb的mRNA信号和P16IR、P21waflIR及PRbIR均是实验组高于对照组,P<005～001,而mp53mRNA信号、PCNAIR、cdk2IR和cdk4IR均是实验组低于对照组,P<005～001。结论 8BrcAMP上调人HXORb44细胞p16、p21wafl、wp53、Rb抑癌基因的表达及P16、P21wafl、PRb蛋白的表达,并下调mp53mRNA、PCNAIR、cdk2IR和cdk4IR的表达,提示8BrcAMP可通过阻抑细胞周期进展相关基因的表达而抑制人HXORb44细胞的生长增殖。
Objective To investigate the effect of 8BrcAMP on tumor suppressor gene expression in human retinoblastoma HXORb44 cells. Methods The mRNA of p16, p21wafl, wp53, mp53 and Rb were detected by dot blot technique. The immunoreactivity (IR) of P16, P21wafl, PRb, cdk2, cdk4 and PCNA were detected by immunohistochemistry. Results The mRNA of p16, p21wafl, wp53 and Rb in human HXORb44 cells and P16IR, P21waflIR and PRbIR were all higher in the experimental group than in the control group, P <005 ~ 001, while the expression of mp53mRNA, PCNAIR, cdk2IR and cdk4IR The experimental group were lower than the control group, P <005 ~ 001. Conclusions 8BrcAMP up-regulated the expression of p16, p21wafl, wp53 and Rb tumor suppressor genes and the expression of P16, P21wafl and PRb in human HXORb44 cells and down-regulated the expressions of mp53mRNA, PCNAIR, cdk2IR and cdk4IR, suggesting that 8BrcAMP may be associated with the suppression of cell cycle progression Gene expression and inhibit the growth and proliferation of human HXORb44 cells.