白细胞介素-33上调A549细胞中黏蛋白基因MUC5AC和MUC5B的表达

Interleukin-33 Promotes the Expression of MUC5AC and MUC5B in A549 Cells

目的:探讨白细胞介素-33(IL-33)对A549细胞中黏蛋白基因表达的影响及其作用机制。方法:体外培养人肺腺癌细胞A549,给予IL-33刺激,通过荧光定量PCR技术(RT-PCR)检测黏蛋白家族基因MUC1,MUC5AC,MUC5B和NF-κB下游基因的mRNA水平,Western Blot检测IκBα的磷酸化水平。随后在IL-33刺激的A549细胞中加入NF-κB抑制剂PDTC,将A549细胞分为对照组,IL-33刺激组,PDTC处理组以及IL-33和PDTC共同处理组,RT-PCR检测黏蛋白家族基因MUC5AC和MUC5B的mRNA水平。结果:IL-33(100ng/ml)处理24h后,A549细胞中MUC5AC和MUC5B的mRNA水平高于对照组(P<0.05),同时NF-κB下游基因(RELA,RELB,NFκB2)的表达也显著高于对照组(P<0.01)。IL33处理A549细胞5min后,IκBα的磷酸化水平显著高于对照组。随后加入NF-κB通路抑制剂PDTC可以有效抑制IL33对MUC5AC基因的激活(P<0.05),而PDTC对MUC5B基因的激活没有影响。结论:IL-33可以通过激活NF-κB信号通路来上调A549细胞中MUC5AC基因的表达,而IL-33上调MUC5B的表达则可能是通过其他信号通路。

Objective:To explore the effect and mechanism of IL-33 on Mucins expression in A549 cells.Methods:Human lung A549 cells cultured in vitro were treated with IL-33,and then the mRNA levels of MUC1,MUC5 AC,MUC5B,and NF-κB downstream genes were examined by reverse transcriptase-polymerase chain reaction(RT-PCR).Western Blot was utilized to evaluate the expression of phosphorylation-level of IκBα.Subsequently,A549 cells were stimulated by IL-33 as well as PDTC,an inhibitor of NF-κB pathway.The cells were then divided into 4groups:a negative control group,an IL-33 treatment group,a PDTC treatment group and a double treatment of IL-33 and PDTC group.The changes of MUC5 AC and MUC5 BmRNA were examined by reverse transcriptase-polymerase chain reaction(RT-PCR).Results:When A549 cells were treated with100ng/ml IL-33 for 24hours,the expression levels of MUC5 AC and MUC5 B mRNA were increased significantly compared with the control group(P<0.05).The mRNA levels of NF-κBdownstream genes(RELA,RELB,NFKB2)were also increased significantly(P<0.01)compared with the control group.When A549 cells were treated with 100ng/ml IL-33 for 5minutes,the phosphorylation-level of IκBαwas increased compared with the control group.And then,NF-κB inhibitor PDTC can effectively inhibit the activation of IL-33 on MUC5AC,while PDTC has no effect on MUC5 B.Conclusion:IL-33 can induce MUC5 AC expression via NF-κB pathway in A549 cells,while it may induce MUC5 Bexpression via other pathways.