摘要
目的探讨Tob与细胞周期蛋白(cyclin)D1的调控关系及其对喉癌细胞增殖的影响。方法荧光定量聚合酶链反应(PCR)检测74例喉癌及相应癌旁正常组织中Tob与cyclinD1的表达;应用U0126处理Hep2细胞,检测处理后细胞中磷酸化的Tob蛋白和Tob蛋白总量的改变,以及cyclinD1的表达量,并用四甲基偶氮唑蓝(MTT)法测定细胞毒性指数;RNA干扰(RNAi)抑制Hep2细胞中Tob蛋白的表达后,检测cyclinD1的表达改变,MTT法测定细胞毒性指数。结果Tob和cyclinD1分别在喉癌组织中表达下降和增加,且二者呈负相关;U0126处理后,磷酸化的Tob减少,Tob蛋白总量无明显改变,cyclinD1表达降低,且细胞增殖下降;siR-NA-Tob转染后可使磷酸化的Tob和Tob总量减少,而cyclinD1表达增加,细胞增殖亦增强。结论Tob和cy-clinD1都与喉癌的发生相关。Tob接受细胞外信号调节激酶(ERK1)的调控,并调节下游cyclinD1基因的表达,影响喉癌细胞的增殖能力,发挥其肿瘤抑制基因的作用。
Objective To explore the relationship between Tob and cyclinD1 in laryngeal cancer,the regulative process between them and their influence on proliferation of Hep2.Methods Seventy-four cases of laryngeal cancer and pericancerous normal tissues were selected to detect the expression of Tob and cyclinD1 by fluorescent quantitative PCR.Hep2 cell was treated by U0126,the expression of phosphor-Tob,Tob and cyclinD1 was detected by western blotting,the proliferation was detected by MTT.Tob was inhibited through RNAi,and the expression of cyclinD1 was detected by western blotting.Then,the proliferation was detected with MTT.Results Compared with pericancerous normal tissues,the expression of Tob and cyclinD1 presented an obvious change in laryngeal cancer;there was a negative relation between them.In Hep2 cell treated by U0126,phosphor-Tob was decreasing,the total Tob was almost the same,and cyclinD1 was decreasing too;the proliferation was found inhibited.In Hep2 cell treated by RNAi,the expression of phosphor-Tob and Tob were decreasing together,and cyclinD1 was incremental;the proliferation enhanced.Conclusion There is a relationship between Tob and cyclinD1 in laryngeal cancer.Tob,acting as a tumor suppressor gene,accepts the regulation of ERK1,controls the expression of cyclinD1 and influences the proliferation of Hep2 cell.
基金
国家自然科学基金(30700980)
