全反式维甲酸对U251细胞系增殖和MAPK信号通路的影响

摘要

目的：探讨全反式维甲酸（all-trans retinoic acid，ATRA）对U251细胞系增殖和MAPK信号通路的影响。方法使用不同浓度的ATRA溶液对脑胶质瘤细胞U251进行孵育，检测ATRA 对U251细胞增殖的影响，并通过qRT-PCR与Western blot方法检测MKPs与MAPK信号通路中各蛋白的变化。结果与对照组相比，ATRA可以有效的抑制脑胶质瘤细胞U251的增殖，并且具有浓度依赖性。qRT-PCR结果显示，采用不同浓度ATRA孵育48h后，MKPs mRNA的表达发生改变，但是MKP-5变化情况与下调67LR表达时不同，说明2种方法对 MAPK信号通路作用的主要区别是对MKP-5的调控。Western blot实验结果表明，ATRA孵育48h后， MAPK信号通路中各蛋白的磷酸化发生变化，说明ATRA通过控制MAPK信号通路中蛋白的磷酸化程度对U251细胞系进行调控。结论维甲酸通过与不同的维甲酸受体相结合，发挥其生理作用，对脑胶质瘤细胞U251进行调控。ATRA可通过降低磷酸化ERK1/2的表达来抑制肿瘤的增殖，且能调控3种蛋白的磷酸化，说明MAPK信号通路在肿瘤增殖过程中发挥重要作用。%Objective To investigate the retinoic acid incubation effect on proliferation of U251 cell line and effect on MAPK signal pathway. Methods ATRA solution of different concentration on the U25 1 glioma cells were incubated,the influence of ATRA on the proliferation of U25 1 cells were detected,and the proteins of MKPs and MAPK signaling pathways were detected by qRT-PCR and Western blot.Using Graph Prism 5 software for quantitative analysis of experimental results.Results Compared with control group,ATRA could effectively inhibit the proliferation of U25 1 glioma cells, in a concentration dependent manner.QRT-PCR results showed that,different concentrations of ATRA after incubation for 48 hours,the expression of MKPs mRNA changed,but the changes of MKP-5 and expression of 67LR was different,explained the main differences between the two methods of the MAPK signaling pathway was the regulation of MKP-5.Western blot results showed that the ATRA,after 48 hours of incubation,the protein MAPK pathway had changed in phosphorylation, which showed that ATRA protein in the MAPK signaling pathway through control of the degree of phosphorylation on U25 1 cell line regulation.Conclusion Retinoic acid and retinoic acid receptor play its physiological effects and regulate human glioma cell line U25 1 proliferation through different combination.Retinoic acid could not only reduce the expression of phosphorylated ERK1/2 to inhibit tumor proliferation,but also regulate three kinds of protein phosphorylation,therefore its mechanism will be more complex,at the same time that the MAPK signaling pathway plays a crucial role in tumor proliferation process.