FOXP3基因沉默抑制肺腺癌A549细胞上皮-间质转化和改善5-氟尿嘧啶耐药性  

FOXP3 gene silencing inhibits epithelial-mesenchymal transformation and improves 5-fluorouracil resistance in lung adenocarcinoma

作　　者：历春[1] 王鹤霏 何程远 房惠 裴怡杰 盖晓东[1] LI Chun;WANG Hefei;HE Chengyuan;FANG Hui;PEI Yijie;GAI Xiaodong(Department of Immunology,School of Basic Medicine,Beihua University,Jilin 132013,China)

机构地区：[1]北华大学基础医学院免疫学教研室,吉林132013 [2]吉林大学白求恩第一医院肿瘤妇科,长春130012

出　　处：《中国免疫学杂志》2022年第10期1212-1217,共6页Chinese Journal of Immunology

基　　金：吉林省自然科学基金项目(YDZJ202101ZYTS089,20190201220JC);吉林省卫生计生委科技项目(2020J017)资助。

摘　　要：目的:探究叉头蛋白3(FOXP3)基因沉默对肺腺癌上皮-间质转化(EMT)及5-氟尿嘧啶(5-FU)耐药性的影响。方法:构建2段特异性针对FOXP3基因的siRNA片段,并将其通过脂质体介导转染至肺腺癌A549细胞,同时设立转染无义序列的阴性对照组。将转染后的A549细胞分为3组:si-NC组(阴性对照)、si-FOXP3-1组和si-FOXP3-2组。qRT-PCR和Western blot检测各组细胞中FOXP3的表达水平,Western blot检测各组细胞中E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)以及N-钙黏蛋白(N-cadherin)表达水平,Transwell检测各组细胞迁移和侵袭能力,ELISA检测各组细胞上清液中基质金属蛋白酶2(MMP-2)和MMP-9蛋白浓度。qRT-PCR和Western blot检测5-FU处理A549细胞后FOXP3表达水平,CCK-8检测各组细胞对5-FU的敏感性,Western blot检测各组细胞中P-糖蛋白(P-gp)表达水平。结果:与si-NC组相比,si-FOXP3-1和si-FOXP3-2组中FOXP3在mRNA和蛋白表达水平均显著降低(P<0.01),EMT相关蛋白Vimentin和N-cadherin表达明显下降而E-cadherin表达明显升高(P<0.01),细胞迁移和侵袭能力明显减弱(P<0.01),MMP-2和MMP-9蛋白浓度明显降低(P<0.01)。5-FU处理的A549细胞中FOXP3表达水平显著高于未经5-FU处理组(P<0.05),与si-NC组相比,si-FOXP3-1组和si-FOXP3-2组细胞对5-FU的敏感性显著增加(P<0.05),P-gp表达显著降低(P<0.01)。结论:FOXP3基因沉默可抑制肺腺癌A549细胞EMT,改善5-FU的耐药性。Objective:To explore the effect of forkhead protein 3(FOXP3)gene silencing on epithelial-mesenchymal transition(EMT)and 5-fluorouracil(5-FU)chemosensitivity in lung adenocarcinoma.Methods:Two FOXP3 gene-specific siRNA were constructed and transfected into lung adenocarcinoma A549 cells by lipofectamine,respectively.At the same time,a nonsense siRNA were used as negative control.The transfected A549 cells were divided into 3 groups:negative control group(si-NC),si-FOXP3-1and si-FOXP3-2 group.qRT-PCR and Western blot were used to detect the expression levels of FOXP3 in each group,Western blot was used to detect the expression levels of E-cadherin,Vimentin and N-cadherin proteins in each group,Transwell was used to detect cell migration and invasion abilities in each group,ELISA was used to detect the concentration of matrix metalloproteinase 2(MMP-2)and MMP-9 in cell supernatant in each group.qRT-PCR and Western blot were used to detect the expression of FOXP3 in A549 cells treated with 5-FU,CCK-8 was used to detect sensitivity of cells to 5-FU and Western blot was used to detect expression of P-glycoprotein(P-gp)in each group.Results:Compared with si-NC group,expression levels of FOXP3 mRNA and protein in si-FOXP3-1 and si-FOXP3-2 groups were both significantly reduced(P<0.01),EMT-related proteins Vimentin and N-cadherin were decreased while E-cadherin were increased(P<0.01),cell migration and invasion abilities were both significantly reduced(P<0.01),concentration of MMP-2 and MMP-9 proteins were both significantly decreased(P<0.01).Expression levels of FOXP3 mRNA and protein in A549cells treated with 5-FU were significantly higher(P<0.05).Compared with si-NC group,sensitivity of cells to 5-FU was increased(P<0.05)and the expression of P-gp was reduced(P<0.01).Conclusion:FOXP3 gene silencing may induce EMT and improve 5-FU resistance in lung adenocarcinoma A549 cells.

关 键 词：叉头蛋白3 肺腺癌 上皮-间质转化 5-氟尿嘧啶 药物敏感性 

分 类 号：R734.2[医药卫生—肿瘤]