摘要
目的:探究SRY相关的高迁移率族盒9(SOX9)通过Wnt/β-连环蛋白(β-catenin)途径促进非小细胞肺癌(NSCLC)A549细胞上皮-间充质转化(EMT)的机制。方法:将A549细胞分为OE-NC组、OE-SOX9组、OE-SOX9+XAV-939组。其中OESOX9组通过转染SOX9pcDNA质粒上调SOX9的表达水平;OE-SOX9+XAV-939组在转染SOX9pcDNA质粒的同时在培养基中加入β-catenin抑制剂XAV-939(1.0μmol/L)。用qPCR检测SOX9 mRNA表达水平,CCK-8法检测A549细胞增殖能力,划痕愈合实验检测A549细胞迁移能力,Transwell小室实验检测A549细胞的侵袭能力,WB实验检测SOX9、β-catenin、E-钙粘蛋白(E-cadherin)、γ-连环蛋白(γ-catenin)、N-钙黏蛋白(N-cadherin)、波形蛋白(vimentin)表达水平。结果:转染后OE-SOX9组和OE-SOX9+XAV-939组的SOX9 mRNA和蛋白的水平显著高于OE-NC组(均P<0.05),且OE-SOX9组和OE-SOX9+XAV-939组比较无显著差异(P>0.05)。OE-SOX9组细胞增殖、侵袭和迁移能力显著高于OE-NC组,而OE-SOX9+XAV-939组显著低于OE-SOX9组(均P<0.05)。OE-SOX9组β-catenin蛋白水平显著高于OE-NC组,而OE-SOX9+XAV-939组β-catenin蛋白的水平低于OE-SOX9组(均P<0.05)。与OE-NC组比较,OE-SOX9组的上皮细胞表型标志物E-cadherin、γ-catenin水平下调并且间充质细胞表型标志物N-cadherin、vimentin上调,而OE-SOX9+XAV-939组E-cadherin、y-catenin高于OE-SOX9组,且N-cadherin、vimentin低于OE-SOX9组(均P<0.05)。结论:SOX9可通过激活Wnt/β-catenin通路促进NSCLCA549细胞的增殖、迁移和EMT。
Objective:To explore the mechanism by which SRY-related high mobility group-box 9(SOX9) promotes the epithelial mesenchymal transition(EMT) of non-small cell lung cancer(NSCLC) A549 cells via the Wnt/β-catenin pathway.Methods:The human NSCLC A549 cell line was divided into three groups:OE-NC group,OE-SOX9 group and OE-SOX9+XAV-939 group.The cells in OESOX9 group were transfected with SOX9 pcDNA plasmid to up-regulate the expression level of SOX9;The cells in OE-SOX9+XAV-939 group were transfected with SOX9 pcDNA plasmid while the β-catenin inhibitor XAV-939(1.0 μmol/L) was added to the medium.qPCR was used to detect SOX9 mRNA levels;CCK-8 was used to examine the proliferation of A549 cells;Wound-healing assay and Transwell chamber assay were used to detect the migration and invasion of A549 cells,respectively;and WB was used to detect protein expressions of SOX9,β-catenin,E-cadherin,y-catenin,N-cadherin and vimentin.Results:The mRNA and protein levels of SOX9 in OE-SOX9 group and OE-SOX9+XAV-939 group were significantly higher than those in the OE-NC group after transfection(all P<0.05),while there was no significant difference between the OE-SOX9 group and the OE-SOX9+XAV-939 group(P>0.05).The proliferation,migration and invasion of cells in OE-SOX9 group were significantly higher than those in OE-NC group;however,those abilities in OE-SOX9+XAV-939 group were significantly lower than those in OE-SOX9 group(all P<0.05).The level of β-catenin protein in OE-SOX9 group was significantly higher than that in the OE-NC group,while the level of β-catenin protein in OE-SOX9+XAV-939 group was lower than that in OE-SOX9 group(all P<0.05).Compared with the OE-NC group,the levels of phenotypic markers of epithelial cells,E-cadherin and y-catenin,were down-regulated,and the phenotypic markers of mesenchymal cells,N-cadherin and vimentin,were up-regulated in cells of OE-SOX9 group;however,E-cadherin and y-catenin were higher,and N-cadherin and vimentin were lower in OE-SOX9+XAV-939 group than those in OE-SOX9 group(all P<0.05).Conclusion:SOX9 could promote proliferation,migration and EMT of NSCLC A549 cells by activating the Wnt/β-catenin pathway.
作者
王秋琼
熊弢
陈江勇
何刚
WANG Qiuqiong;XIONG Tao;CHEN Jiangyong;HE Gang(Department of Respiratory,Yongchuan Hospital Affiliated to Chongqing Medical University,Chongqing 402160,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2019年第12期1345-1349,共5页
Chinese Journal of Cancer Biotherapy
