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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Rel A、Rel B mRNA,细胞因子LTα、LTβ、LIGHT、TNFα、IL-6、IL-1βmRNA和增殖相关基因细胞周期素D1(Cyclin D1)、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,Rel A mRNA表达上调2.5倍(P=0.003),TNFα、IL-1β、Cyclin D1、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Rel A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Cyclin D1、Survivin的表达,但在细胞增殖水平上并未见明显效应。
Objective To investigate the effect of activation of lymphotoxin β receptor (LTβR) on the nuclear factor-kappa B (NF-κB) signaling pathway and cell proliferation in bladder cancer cell line 5637. Methods Bladder cancer cell line 5637 was used as a study object, and the activation of LTβR was induced by the ligand lymphotoxin α1β2 (LTα1β2). qRT-PCR was used to detect the expression of Rel A, Rel B mRNA, cytokines LTα, LTβ, LIGHT, TNFα, IL-6 and IL-1βmRNA and proliferation related genes Cyclin D1 and Survivin ) mRNA was detected by Western blot. The activation of NF-κB signaling pathway was detected by WB method. The proliferation of cells was detected by CCK-8 assay. Results After activation of LTβR, mRNA expression of Rel A was up-regulated by 2.5 folds (P = 0.003), while mRNA expression of TNFα, IL-1β, Cyclin D1 and Survivin increased to different extents (all P < (P <0.05). The results of WB showed that the expression of p-p65 was up-regulated (P> 0.05). Compared with the control group, there was no significant difference in the proliferation of cells ). Conclusion Activation of LTβR can promote the expression and activation of Rel A, a key member of the NF-κB signaling pathway. It may be involved in the formation of inflammatory microenvironment of bladder cancer by up-regulating the expression of pro-inflammatory cytokines TNFα and IL-1β. Gene Cyclin D1, Survivin expression, but no significant effect on the level of cell proliferation.