Lymphotoxin-beta receptor activation in bladder cancer cells affecting NF-κB signaling and proliferation
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摘要: 目的 研究淋巴毒素β受体(LTβR)的活化对膀胱癌细胞株5637核转录因子-κB(NF-κB)信号通路及细胞增殖的影响。 方法 以膀胱癌细胞株5637为研究对象,以配体淋巴毒素α1β2(LTα1β2)诱导LTβR信号活化。qRT-PCR检测NF-κB信号通路RelA、RelB mRNA,细胞因子LTα、LTβ、LIGHT、TNFα、IL-6、IL-1β mRNA和增殖相关基因细胞周期素D1(CyclinD1)、生存素(Survivin) mRNA的表达水平;WB法检测NF-κB经典信号通路活化状态;CCK-8法检测细胞增殖水平。 结果 LTβR活化后,RelA mRNA表达上调2.5倍(P=0.003),TNFα、IL-1β、CyclinD1、Survivin mRNA出现不同程度的上调(均P<0.05),随着LTβR表达下调,以上基因mRNA表达也下降(均P<0.05);WB结果显示活化标志蛋白p-p65表达出现升高趋势(P>0.05);与对照组相比,细胞增殖活性的差异未见统计学意义(P>0.05)。 结论 活化LTβR可促进NF-κB经典信号通路主要成员RelA的表达和活化,并有可能通过上调促炎细胞因子TNFα、IL-1β的表达参与膀胱癌炎性微环境的形成;有利促增殖相关基因CyclinD1、Survivin的表达,但在细胞增殖水平上并未见明显效应。Abstract: Objective To study the effect of lymphotoxin-beta receptor (LTβR) activation on NF-κB signaling and proliferation of bladder cancer cells. Methods The bladder cancer cells 5637 were cultured in our study,the activation of LTβR was induced by lymphotoxin α1β2(LTα1β2),a functional ligand.The qRT-PCR was applied to detect the mRNA expression of NF-κB family main members (RelA,RelB),cytokines including LTα,LTβ,LIGHT,TNFα,IL-6,IL-1β,and the proliferation-related genes including CyclinD1 and Survivin.The level of phospho-p65(p-p65) was determined by western blot.Cell viability was tested by CCK-8. Results After activation of LTβR,RelA mRNA was up-regulated about 2.5 times (P=0.003),TNFα,IL-1β,CyclinD1 and Survivin mRNA expressions were up-regulated significantly (all P<0.05).Along with the down-regulated expression of LTβR,the mRNA expression of the above-mentioned gene were also decreased (P all<0.05).WB determined that there was an up-regulated trend of p-p65 after activation of LTβR (all P>0.05).There were no significant differences in the proliferation of bladder cancer cells compared to the control (P both>0.05). Conclusion Activation of LTβR can promote the expression of NF-κB family member RelA and the activation of the classical NF-κB signaling.It might be a participant in the emergence of inflammatory microenvironment through up-regulating the expression of TNFα and IL-1β mRNA;Though CyclinD1 and Survivin have been up-regulated,but LTβR has no effect on the proliferation level of bladder cancer cell.
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