膀胱癌组织黑色素瘤抗原-A3 的表达及其对 核因子Kappa B 信号通路的影响

Expression of MAGE-A3 in bladder carcinoma and regulatory role in NF-κB Signaling pathway-related inflammatory factors

摘要:

目的 探讨膀胱癌组织中黑色素瘤抗原-A3（MAGE-A3）的表达变化及其对核因子Kappa B （NF-κB）信号通路相关炎症因子表达的影响。方法 依照纳入与排除标准，收集该院膀胱癌患者120 例。 实时荧光定量PCR（qRT-PCR）和Western blot 检测膀胱癌组织中MAGE-A3 mRNA 和蛋白的表达变化； ELISA 检测膀胱癌组织中NF-κB 信号通路相关炎症因子如肿瘤坏死因子α（TNF-α）和白介素1β（IL- 1β）的表达变化；分析MAGE-A3 阳性率和炎症因子含量与年龄、性别、肿瘤组织类型及肿瘤分期等的关系； 培养膀胱癌细胞系T24，采用小干扰RNA（siRNA）敲减MAGE-A3 的表达后，检测NF-κB P65 磷酸化水 平变化及TNF-α 和IL-1β 的表达变化。结果 膀胱癌组织中MAGE-A3 mRNA、蛋白及炎症因子的表达 水平高于正常膀胱组织，差异有统计学意义（P <0.01）；MAGE-A3 阳性率及患者肿瘤组织中炎症因子含量 与一般临床资料无相关，而与膀胱癌分期呈正相关；siRNA 敲减MAGE-A3 的表达后，NF-κB p65 磷酸化 水平及TNF-α 和IL-1β 的表达水平均降低，差异有统计学意义（均P < 0.01）。结论 MAGE-A3 可能通过 NF-κB 信号通路调控膀胱癌细胞炎症状态，促进膀胱癌的发生发展。

Abstract:

Objective To investigate the expression of melanoma-associated antigen-A3 (MAGE-A3) in bladder carcinoma and its regulatory effect on the expression of Nuclear Factor kappa B (NF-κB) signaling pathway related inflammatory factors. Methods Totally 120 cases of bladder cancer were involved in this study based on inclusion and exclusion criteria. RT-qPCR and Western blot were used to detect the expression of MAGE-A3 in bladder cancer tissues. ELISA assay was performed to detect the expression of NF-κB signaling related factors such as tumor necrosis factor alpha (TNF-α) and interleukin 1β (IL-1β). Association between MAGE-A3 positive rate or inflammatory factor content and age, sex, tumor type and tumor staging were determined. MAGE-A3 was knocked down by small interfering RNA (siRNA) in bladder cancer cell line T24. Results Expression of MAGE-A3 and inflammatory factor in bladder cancer tissues was significantly higher than that in normal bladder tissues (P < 0.01). The positive rate of MAGE-A3 and the content of inflammatory factors in tumor tissue were not correlated with general clinical data while positively associated with the stage of bladder cancer. SiRNA knockdown of MAGE-A3 induced significant reduction of NF-κB p65 phosphorylation levels and TNF-α and IL-1β expression levels ((P < 0.01). Conclusion MAGE-A3 regulates the inflammatory response and promote the development of bladder cancerthrough NF-κB signaling pathway.