摘要
目的:研究肿瘤坏死因子α(tumor necrosis factor-alpha, TNF-α)在非小细胞肺癌(non-small cell lung cancer, NSCLC)中对NF-κB/PXR信号通路和去SUMO化(small ubiquitin-like modifier,SUMO)修饰的调控作用。方法:培养人正常肺上皮细胞系BEAS-2B和人非小细胞肺癌细胞系A549,在两种细胞系中加入人重组肿瘤坏死因子α(recombinant human tumor necrosis factor-alpha, rhTNF-α),采用实时荧光定量PCR(qRT-PCR)方法检测核转录因子-κB (nuclear transcription factor-κB, NF-κB)、孕烷X受体(Pregnane X receptor, PXR)和多药耐药基因-1(multidrug resistance-1, MDR-1)以及包括SENP1、SENP2和SENP3在内的SUMO特异性蛋白酶(SUMO-specific proteases, SENPs)的m RNA水平的变化;采用细胞免疫组织化学染色的方法验证NF-κB、PXR和SENP1在蛋白水平的变化;采用Western blot方法检测磷酸化NF-κB的表达改变。结果:基础状态下,A549细胞系与BEAS-2B细胞系中NF-κB表达水平差异无统计学意义(P=0.745);加入TNF-α诱导后,A549细胞系和BEAS-2B细胞系中NF-κB的m RNA和蛋白水平以及磷酸化NF-κB表达水平升高,A549细胞系中的NF-κB及磷酸化NF-κB表达水平均明显高于BEAS-2B细胞系(P<0.05)。基础状态下,A549细胞系中PXR、MDR-1及SENP1的表达明显高于BEAS-2B细胞系,SENP2及SENP3的表达明显低于BEAS-2B细胞系(P<0.05);TNF-α诱导后,A549细胞系PXR、MDR-1、SENP1、SENP2和SENP3的表达均降低,但BEAS-2B细胞系中以上指标表达均升高(P<0.05)。结论:炎症因子TNF-α可以诱导NSCLC细胞系中NF-κB表达上调,PXR、MDR-1、SENP1、SENP2和SENP3的表达下调,提示TNF-α可能通过NF-κB/PXR炎症通路参与肿瘤的发生过程,并且,去SUMO化修饰可能参与TNF-α对NF-κB/PXR通路的调控作用。
Objective: To explore the regulation of TNF-α(tumor necrosis factor-alpha) on NF-κB/PXR signaling pathway and deSUMOylation modulation in non-small cell lung cancer. Methods: Human normal lung epithelial BEAS-2 B cell line and human non-small cell lung cancer A549 cell line were used to investigate the changes in m RNA of NF-κB(nuclear transcription factor-κB),PXR(Pregnane X receptor), MDR-1(multidrug resistance-1) and deSUMOylation enzymes including SENP1, SENP2, and SENP3 in recombinant human TNF-α(rhTNF-α)-induced NSCLC A549 and BEAS-2 B cell line by quantitative RT-PCR(qRT-PCR). Cellular immunochemical staining was used to verify the changes in protein levels of NF-κB, PXR and SENP1. Changes of phosphorylated NF-κB were detected by Western blot. Results: The baseline expression of NF-κB m RNA was similar in both A549 cell line and control(P=0.745). Increased expression of NF-κB in m RNA and protein levels as well as phosphorylated NF-κB after TNF-α treatment was found in A549 cell line and normal control. However, the expression in A549 cells was significantly higher than BEAS-2 B cells(P<0.05).The baseline expression of PXR, MDR-1 and SENP1 is higher in A549 cells, but the expression of SENP2 and SENP3 was much lower compared to BEAS-2 B cells(P<0.05). The TNF-α treatment decreased the expression of PXR, MDR-1, SENP1, SENP2 and SENP3 in A549 cells, but increased the expression in BEAS-2 B cells(P<0.05). Conclusions: TNF-α is involved in the contribution of NF-κB/PXR signaling pathway to carcinogenesis in NSCLC cell line. In addition, de-SUMOylation may play a role in the regulation of TNF-α on NF-κB/PXR pathway.
作者
于雪姣
卫红军
张芳
周磊
黄维清
YU Xue-jiao;WEI Hong-jun;ZHANG Fang;ZHOU Lei;HUANG Wei-qing(Department of Pathology,Basic Medical College,Qingdao University,Qingdao,Shandong,266071,China;Department of Pathology,Qingdao Municipal Hospital,Qingdao,Shandong,266071,China)
出处
《现代生物医学进展》
CAS
2020年第17期3221-3226,共6页
Progress in Modern Biomedicine
基金
山东省自然科学基金项目(ZR2016HM28)
青岛市医疗卫生重点学科建设项目(青卫科教字(2017)9号)。
