IL-33促进非小细胞肺癌EMT的研究

Study of IL-33 promoting EMT in non-small cell lung cancer

目的探讨IL-33参与非小细胞肺癌(non-small cell lung cancer,NSCLC)上皮细胞-间充质转化(epithelial-mesenchymal transition,EMT)的过程及临床意义。方法生物信息学分析IL-33在非小细胞肺癌中的生物学行为。https://www.ncbi.nlm.nih.gov/geo/网站下载非小细胞肺癌芯片GSE5843和GSE102287进行生存分析。免疫组织化学评估非小细胞肺癌中蛋白IL-33、ST2的表达,免疫荧光法检测E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)的组织定位和分布。体外CCK-8法检测IL-33刺激下A549、SK-MES-1细胞的细胞活力,划痕实验和Transwell实验检测IL-33促进A549、SK-MES-1细胞迁移和侵袭能力。免疫印迹实验检测最佳浓度IL-33处理下ST2、E-cadherin、波形蛋白(Vimentin)、基质金属蛋白酶-9(Matrixmetalloproteinase-9,MMP-9)蛋白的表达。结果生物信息学分析发现IL-33可通过EMT参与NSCLC的进展和转移,生存分析提示IL-33与非小细胞肺癌预后有关,IL-33高表达者,预后差(P<0.05)。免疫组化结果显示与癌旁组织相比,非小细胞肺癌中IL-33、ST2高表达,E-cadherin表达下降,Vimentin明显增多(P<0.05),且在肺腺癌与肺鳞癌之间的表达无明显差异(P>0.05)。IL-33可促进A549、SK-MES-1细胞增殖、迁移和侵袭。与HPAEpiC细胞相比,A549、SK-MES-1细胞高表达ST2、Vimentin、MMP-9、E-cadherin(P<0.05),IL-33刺激下A549、SK-MES-1细胞ST2、Vimentin、MMP-9表达明显升高,E-cadherin明显下降(P<0.05)。结论非小细胞肺癌中IL-33通过ST2促进EMT过程,促进非小细胞肺癌进展。

Objective To investigate IL-33 in epithelial mesenchymal transition(EMT)in non-small cell lung cancer(NSCLC)and its clinical significance.Methods The biological behavior of IL-33 in non-small cell lung cancer was assessed by bioinformatics analysis.NSCLC chips GSE5843 and GSE102287 were download from https://www.ncbi.nlm.nih.gov/geo/for survival analysis.Immunohistochemistry was used to evaluate the expression of protein IL-33 and ST2,and immunofluorescence was used to detect the tissue localization and distribution of E-cadherin and Vimentin in non-small cell lung cancer.In vitro,CCK-8 was used to detect the cell viability of A549 and SK-MES-1 cells stimulated by IL-33,and scratch test and Transwell test were adopted to examine the migration and invasion of A549 and SK-MES-1 cells stimulated by IL-33.Western blot was used to detect the expression of ST2,E-cadherin,Vimentin,and Matrix metalloproteinase-9(MMP-9)proteins under the optimal concentration of IL-33.Results Bioinformatics analysis revealed that IL-33 can participate in the progression and metastasis of NSCLC through EMT.Survival analysis suggested that IL-33 was associated with the prognosis of non-small cell lung cancer,and those with high expression of IL-33 had poor prognosis(P<0.05).Immunohistochemical results showed that compared with adjacent tissues,non-small cell lung cancer showed high expression of IL-33 and ST2,decreased expression of E-cadherin and significantly increased expression of Vimentin(P<0.05),but there was no significant difference between lung adenocarcinoma and squamous cell lung carcinoma(P>0.05).In vitro,IL-33 could promote the proliferation,migration and invasion of A549 and SK-MES-1 cells.Compared with HPAEpiC cells,A549 and SK-MES-1 cells showed high expression of ST2,Vimentin,MMP-9,and E-cadherin(P<0.05).Under IL-33 stimulation,the expression of ST2,Vimentin,and MMP-9 in A549 and SK-MES-1 cells was significantly increased,while E-cadherin was significantly decreased(P<0.05).Conclusion Facilitating the EMT process through ST2,IL-33 can promote non-small cell lung cancer progression.